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J.; Al‐Nakib, W. title: Direct detection of rhinoviruses by an enzyme‐linked immunosorbent assay date: 2005-12-07 journal: J Med Virol DOI: 10.1002/jmv.1890230211 sha: doc_id: 9922 cord_uid: t1hoox6e file: cache/cord-007497-nn5l5rai.json key: cord-007497-nn5l5rai authors: Garcia-Sanchez, J.; Corral, C.; Halaihel, N.G.; Simon, M.C.; Alonso, J.L.; Muzquiz, J.L.; Ortega, C.; Girones, O. title: Survey of rotavirus infection in a dairy herd: comparison between polycrylamide gel electrophoresis and two commercial tests date: 2002-11-13 journal: Vet Microbiol DOI: 10.1016/0378-1135(93)90057-e sha: doc_id: 7497 cord_uid: nn5l5rai file: cache/cord-001446-mpuovmeb.json key: cord-001446-mpuovmeb authors: Bratcher, Preston E.; Gaggar, Amit title: Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA date: 2014-11-03 journal: PLoS One DOI: 10.1371/journal.pone.0111466 sha: doc_id: 1446 cord_uid: mpuovmeb file: cache/cord-003623-n01rgqyv.json key: cord-003623-n01rgqyv authors: Schuh, Amy J.; Amman, Brian R.; Sealy, Tara S.; Flietstra, Timothy D.; Guito, Jonathan C.; Nichol, Stuart T.; Towner, Jonathan S. title: Comparative analysis of serologic cross-reactivity using convalescent sera from filovirus-experimentally infected fruit bats date: 2019-04-30 journal: Sci Rep DOI: 10.1038/s41598-019-43156-z sha: doc_id: 3623 cord_uid: n01rgqyv file: cache/cord-007495-gpz4gkv3.json key: cord-007495-gpz4gkv3 authors: Weiss, M.; Steck, F.; Kaderli, R.; Horzinek, M.C. title: Antibodies to berne virus in horses and other animals date: 2002-11-13 journal: Vet Microbiol DOI: 10.1016/0378-1135(84)90014-2 sha: doc_id: 7495 cord_uid: gpz4gkv3 file: cache/cord-011212-ovjdzyxv.json key: cord-011212-ovjdzyxv authors: Pan, Qing; Wang, Jing; Gao, Yulong; Cui, Hongyu; Liu, Changjun; Qi, Xiaole; Zhang, Yanping; Wang, Yongqiang; Li, Kai; Gao, Li; Wang, Xiaomei title: Development and application of a novel ELISA for detecting antibodies against group I fowl adenoviruses date: 2019-12-14 journal: Appl Microbiol Biotechnol DOI: 10.1007/s00253-019-10208-3 sha: doc_id: 11212 cord_uid: ovjdzyxv file: cache/cord-003859-k8wfyj9b.json key: cord-003859-k8wfyj9b authors: Paweska, Janusz T.; Moolla, Naazneen; Storm, Nadia; Msimang, Veerle; Conteh, Ousman; Weyer, Jacqueline; van Vuren, Petrus Jansen title: Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera date: 2019-07-24 journal: Viruses DOI: 10.3390/v11080678 sha: doc_id: 3859 cord_uid: k8wfyj9b file: cache/cord-012891-heqsfzkm.json key: cord-012891-heqsfzkm authors: Blanco Vázquez, Cristina; Alonso-Hearn, Marta; Juste, Ramón A.; Canive, María; Iglesias, Tania; Iglesias, Natalia; Amado, Javier; Vicente, Fernando; Balseiro, Ana; Casais, Rosa title: Detection of latent forms of Mycobacterium avium subsp. paratuberculosis infection using host biomarker-based ELISAs greatly improves paratuberculosis diagnostic sensitivity date: 2020-09-03 journal: PLoS One DOI: 10.1371/journal.pone.0236336 sha: doc_id: 12891 cord_uid: heqsfzkm file: cache/cord-007644-7bsixsgd.json key: cord-007644-7bsixsgd authors: Chirnside, E.D.; Francis, P.M.; De Vries, A.A.F.; Sinclaira, R.; Mumford, J.A. title: Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus date: 2000-04-04 journal: J Virol Methods DOI: 10.1016/0166-0934(95)00020-u sha: doc_id: 7644 cord_uid: 7bsixsgd file: cache/cord-009877-3cyz6o9c.json key: cord-009877-3cyz6o9c authors: Barclay, Wendy S.; Callow, Kathleen A.; Sergeant, Marianne; Ai‐Nakib, Widad title: Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions date: 2005-12-07 journal: J Med Virol DOI: 10.1002/jmv.1890250411 sha: doc_id: 9877 cord_uid: 3cyz6o9c file: cache/cord-014462-11ggaqf1.json key: cord-014462-11ggaqf1 authors: nan title: Abstracts of the Papers Presented in the XIX National Conference of Indian Virological Society, “Recent Trends in Viral Disease Problems and Management”, on 18–20 March, 2010, at S.V. University, Tirupati, Andhra Pradesh date: 2011-04-21 journal: Indian J Virol DOI: 10.1007/s13337-011-0027-2 sha: doc_id: 14462 cord_uid: 11ggaqf1 file: cache/cord-009784-kxa68zbc.json key: cord-009784-kxa68zbc authors: Bolton, Jessica S.; Chaudhury, Sidhartha; Dutta, Sheetij; Gregory, Scott; Locke, Emily; Pierson, Tony; Bergmann-Leitner, Elke S. title: Comparison of ELISA with electro-chemiluminescence technology for the qualitative and quantitative assessment of serological responses to vaccination date: 2020-04-17 journal: Malar J DOI: 10.1186/s12936-020-03225-5 sha: doc_id: 9784 cord_uid: kxa68zbc file: cache/cord-011840-neowfhwg.json key: cord-011840-neowfhwg authors: Liu, Weixiao; Liu, Xuri; Liu, Chao; Zhang, Zhe; Jin, Wujun title: Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops date: 2020-03-05 journal: Biotechnol Lett DOI: 10.1007/s10529-020-02854-9 sha: doc_id: 11840 cord_uid: neowfhwg file: cache/cord-281081-rifr5uub.json key: cord-281081-rifr5uub authors: Deng, Junhua; Jin, Yipeng; Liu, Yuxiu; Sun, Jie; Hao, Liying; Bai, Jingjing; Huang, Tian; Lin, Degui; Jin, Yaping; Tian, Kegong title: Serological survey of SARS‐CoV‐2 for experimental, domestic, companion and wild animals excludes intermediate hosts of 35 different species of animals date: 2020-05-07 journal: Transbound Emerg Dis DOI: 10.1111/tbed.13577 sha: doc_id: 281081 cord_uid: rifr5uub file: cache/cord-013348-lsksys56.json key: cord-013348-lsksys56 authors: Goto, Keiko; Yamaoka, Yutaro; Khatun, Hajera; Miyakawa, Kei; Nishi, Mayuko; Nagata, Noriko; Yanaoka, Toshikazu; Kimura, Hirokazu; Ryo, Akihide title: Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 date: 2020-09-19 journal: Microorganisms DOI: 10.3390/microorganisms8091437 sha: doc_id: 13348 cord_uid: lsksys56 file: cache/cord-255390-dvp0luxe.json key: cord-255390-dvp0luxe authors: Jones, R. 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Evolutionary implications date: 2009-03-20 journal: Virus Res DOI: 10.1016/j.virusres.2009.02.019 sha: doc_id: 257715 cord_uid: pbcr81qm file: cache/cord-014965-efmozngq.json key: cord-014965-efmozngq authors: nan title: Infectious diseases other than CMV (1st Section) date: 2001-06-11 journal: Bone Marrow Transplant DOI: 10.1038/sj.bmt.1702942 sha: doc_id: 14965 cord_uid: efmozngq file: cache/cord-262640-4vr4cm1s.json key: cord-262640-4vr4cm1s authors: Nguyen, N. N.; Mutnal, M. B.; Gomez, R. R.; Pham, H. N.; Nguyen, L. T.; Koss, W.; Rao, A.; Arroliga, A. C.; Wang, L.; Wang, D.; Hua, Y.; Powell, P. R.; Chen, L.; McCormack, C.; Linz, W. J.; Mohammad, A. A. title: Correlation of ELISA based with random access serologic immunoassays for identifying adaptive immune response to SARS-CoV-2 date: 2020-07-08 journal: nan DOI: 10.1101/2020.07.06.20145938 sha: doc_id: 262640 cord_uid: 4vr4cm1s file: cache/cord-275793-k0uvqcmp.json key: cord-275793-k0uvqcmp authors: Xia, Hongyan; Liu, Lihong; Nordengrahn, Ann; Kiss, István; Merza, Malik; Eriksson, Ronnie; Blomberg, Jonas; Belák, Sándor title: A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies date: 2010-04-18 journal: J Virol Methods DOI: 10.1016/j.jviromet.2010.04.009 sha: doc_id: 275793 cord_uid: k0uvqcmp file: cache/cord-261372-xjbs09gi.json key: cord-261372-xjbs09gi authors: Sozzi, Enrica; Luppi, Andrea; Lelli, Davide; Martin, Ana Moreno; Canelli, Elena; Brocchi, Emiliana; Lavazza, Antonio; Cordioli, Paolo title: Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus date: 2010-02-28 journal: Research in Veterinary Science DOI: 10.1016/j.rvsc.2009.05.009 sha: doc_id: 261372 cord_uid: xjbs09gi file: cache/cord-276989-441aclcc.json key: cord-276989-441aclcc authors: Liu, Jianbo; Gao, Ran; Shi, Hongyan; Cong, Guangyi; Chen, Jianfei; Zhang, Xin; Shi, Da; Cao, Liyan; Wang, Xiaobo; Zhang, Jialin; Ji, Zhaoyang; Jing, Zhaoyang; Feng, Li title: Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum date: 2020-03-12 journal: J Virol Methods DOI: 10.1016/j.jviromet.2020.113855 sha: doc_id: 276989 cord_uid: 441aclcc file: cache/cord-277265-p8pns7r9.json key: cord-277265-p8pns7r9 authors: Malik, Yashpal Singh; Verma, Atul; Kumar, Naveen; Deol, Pallavi; Kumar, Deepak; Ghosh, Souvik; Dhama, Kuldeep title: Biotechnological innovations in farm and pet animal disease diagnosis date: 2019-09-20 journal: Genomics and Biotechnological Advances in Veterinary, Poultry, and Fisheries DOI: 10.1016/b978-0-12-816352-8.00013-8 sha: doc_id: 277265 cord_uid: p8pns7r9 file: cache/cord-264936-3posyr5n.json key: cord-264936-3posyr5n authors: Mohammadzadeh, Sara; Khabiri, Alireza; Roohvand, Farzin; Memarnejadian, Arash; 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Yang, Ling; Zhang, Ting; Yin, Ye; Cui, Xiaodai; Tang, Xiangjun; Wang, Luoping; He, Bo; Ma, Lianhua; Lei, Tingting; Zeng, Changqing; Fang, Jianqiu; Yu, Jun; Wang, Jian; Yang, Huanming; West, Matthew B; Bhatnagar, Aruni; Lu, Youyong; Xu, Ningzhi; Liu, Siqi title: Assessment of Immunoreactive Synthetic Peptides from the Structural Proteins of Severe Acute Respiratory Syndrome Coronavirus date: 2003-12-01 journal: Clin Chem DOI: 10.1373/clinchem.2003.023184 sha: doc_id: 262268 cord_uid: gm99cadh file: cache/cord-273361-i5v5rz4x.json key: cord-273361-i5v5rz4x authors: Chen, Tsu-Han; Lee, Fan; Lin, Yeou-Liang; Pan, Chu-Hsiang; Shih, Chia-Ni; Lee, Ming-Chang; Tsai, Hsiang-Jung title: Development of a Luminex assay for the detection of swine antibodies to non-structural proteins of foot-and-mouth disease virus date: 2013-10-31 journal: Journal of Immunological Methods DOI: 10.1016/j.jim.2013.08.002 sha: doc_id: 273361 cord_uid: i5v5rz4x file: cache/cord-294478-3ickafd3.json key: cord-294478-3ickafd3 authors: Kapil, Sanjay; 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diagnostic accuracies of rapid serological tests and ELISA to molecular diagnostics in patients with suspected COVID-19 presenting to the hospital date: 2020-06-02 journal: Clin Microbiol Infect DOI: 10.1016/j.cmi.2020.05.028 sha: doc_id: 344581 cord_uid: h7ikjgic file: cache/cord-340960-abanr641.json key: cord-340960-abanr641 authors: Brigger, D.; Horn, M.P.; Pennington, L.F.; Powell, A.E.; Siegrist, D.; Weber, B.; Engler, O.; Piezzi, V.; Damonti, L.; Iseli, P.; Hauser, C.; Froehlich, T.K.; Villiger, P.M.; Bachmann, M.F.; Leib, S.L.; Bittel, P.; Fiedler, M.; Largiadèr, C.; Marschall, J.; Stalder, H.; Kim, P.S.; Jardetzky, T.S.; Eggel, A.; Nagler, M. title: Accuracy of serological testing for SARS‐CoV‐2 antibodies: first results of a large mixed‐method evaluation study date: 2020-09-30 journal: Allergy DOI: 10.1111/all.14608 sha: doc_id: 340960 cord_uid: abanr641 file: cache/cord-346320-ysgz6adr.json key: cord-346320-ysgz6adr authors: Arabi, Yaseen M.; Hajeer, Ali H.; Luke, Thomas; Raviprakash, Kanakatte; Balkhy, Hanan; Johani, Sameera; Al-Dawood, Abdulaziz; Al-Qahtani, Saad; Al-Omari, Awad; Al-Hameed, Fahad; Hayden, Frederick G.; Fowler, Robert; Bouchama, Abderrezak; Shindo, Nahoko; Al-Khairy, Khalid; Carson, Gail; Taha, Yusri; Sadat, Musharaf; Alahmadi, Mashail title: Feasibility of Using Convalescent Plasma Immunotherapy for MERS-CoV Infection, Saudi Arabia date: 2016-09-17 journal: Emerg Infect Dis DOI: 10.3201/eid2209.151164 sha: doc_id: 346320 cord_uid: ysgz6adr file: cache/cord-351498-bmq6zcb0.json key: cord-351498-bmq6zcb0 authors: Martínez-Sernández, Victoria; Perteguer, María J.; Hernández-González, Ana; Mezo, Mercedes; González-Warleta, Marta; Orbegozo-Medina, Ricardo A.; Romarís, Fernanda; Paniagua, Esperanza; Gárate, Teresa; Ubeira, Florencio M. title: Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis date: 2018-03-21 journal: Parasitol Res DOI: 10.1007/s00436-018-5809-7 sha: doc_id: 351498 cord_uid: bmq6zcb0 file: cache/cord-348161-757c51xw.json key: cord-348161-757c51xw authors: Petrosova, A.; Konry, T.; Cosnier, S.; Trakht, I.; Lutwama, J.; Rwaguma, E.; Chepurnov, A.; Mühlberger, E.; Lobel, L.; Marks, R.S. title: Development of a highly sensitive, field operable biosensor for serological studies of Ebola virus in central Africa date: 2007-03-26 journal: Sens Actuators B Chem DOI: 10.1016/j.snb.2006.07.005 sha: doc_id: 348161 cord_uid: 757c51xw file: cache/cord-353614-z4fpy607.json key: cord-353614-z4fpy607 authors: Kusano, Nario; Hirashima, Keita; Kuwahara, Minoru; Narahara, Kenji; Imamura, Tadashi; Mimori, Tomohiro; Nakahira, Ken; Torii, Kuniaki title: Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies date: 2007-02-16 journal: J DOI: 10.1007/s10327-006-0316-6 sha: doc_id: 353614 cord_uid: z4fpy607 file: cache/cord-353190-7qcoxl81.json key: cord-353190-7qcoxl81 authors: Nicklas, Werner; Bleich, André; Mähler, Michael title: Viral Infections of Laboratory Mice date: 2012-05-17 journal: The Laboratory Mouse DOI: 10.1016/b978-0-12-382008-2.00019-2 sha: doc_id: 353190 cord_uid: 7qcoxl81 file: cache/cord-019347-tj3ye1mx.json key: cord-019347-tj3ye1mx authors: nan title: ABSTRACT BOOK date: 2010-02-19 journal: Ann Allergy Asthma Immunol DOI: 10.1016/s1081-1206(10)61294-x sha: doc_id: 19347 cord_uid: tj3ye1mx file: cache/cord-022501-9wnmdvg5.json key: cord-022501-9wnmdvg5 authors: nan title: P1460 – P1884 date: 2015-12-28 journal: Clin Microbiol Infect DOI: 10.1111/j.1470-9465.2006.12_4_1431.x sha: doc_id: 22501 cord_uid: 9wnmdvg5 file: cache/cord-022888-dnsdg04n.json key: cord-022888-dnsdg04n authors: nan title: Poster Sessions date: 2009-08-19 journal: Eur J Immunol DOI: 10.1002/eji.200990224 sha: doc_id: 22888 cord_uid: dnsdg04n file: cache/cord-023095-4dannjjm.json key: cord-023095-4dannjjm authors: nan title: Research Abstract Program of the 2011 ACVIM Forum Denver, Colorado, June 15–18, 2011 date: 2011-05-03 journal: J Vet Intern Med DOI: 10.1111/j.1939-1676.2011.0726.x sha: doc_id: 23095 cord_uid: 4dannjjm file: cache/cord-015021-pol2qm74.json key: cord-015021-pol2qm74 authors: nan title: Third International Congress on the Immune Consequences of Trauma, Shock and Sepsis —Mechanisms and Therapeutic Approaches date: 1994 journal: Intensive Care Med DOI: 10.1007/bf02258437 sha: doc_id: 15021 cord_uid: pol2qm74 file: cache/cord-031907-ilhr3iu5.json key: cord-031907-ilhr3iu5 authors: nan title: ISEV2020 Abstract Book date: 2020-07-15 journal: nan DOI: 10.1080/20013078.2020.1784511 sha: doc_id: 31907 cord_uid: ilhr3iu5 file: cache/cord-022940-atbjwpo5.json key: cord-022940-atbjwpo5 authors: nan title: Poster Sessions date: 2016-09-07 journal: FEBS J DOI: 10.1111/febs.13808 sha: doc_id: 22940 cord_uid: atbjwpo5 file: cache/cord-015394-uj7fe5y6.json key: cord-015394-uj7fe5y6 authors: nan title: Scientific Abstracts date: 2008-12-23 journal: Reprod Sci DOI: 10.1177/19337191080150020102 sha: doc_id: 15394 cord_uid: uj7fe5y6 Reading metadata file and updating bibliogrpahics === updating bibliographic database Building study carrel named keyword-elisa-cord === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 94844 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95786 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 94058 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95519 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96163 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95982 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95840 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96636 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95484 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95744 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95832 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95914 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96878 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96260 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 94491 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96206 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96282 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 94989 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95069 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96875 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96581 Aborted $FILE2BIB "$FILE" > "$OUTPUT" parallel: Warning: No more processes: Decreasing number of running jobs to 95. parallel: Warning: Raising ulimit -u or /etc/security/limits.conf may help. === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96102 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96414 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 95550 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96599 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96618 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96684 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96439 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 98104 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97669 Aborted $FILE2BIB "$FILE" > "$OUTPUT" parallel: Warning: No more processes: Decreasing number of running jobs to 94. parallel: Warning: Raising ulimit -u or /etc/security/limits.conf may help. === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96222 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96490 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96547 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96847 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96849 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97073 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97271 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 98115 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 98270 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 505 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97459 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97970 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 96700 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97488 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97668 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97896 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === id: cord-007480-grndfx7b author: Koopmans, M. title: Seroepidemiology of Breda virus in cattle using ELISA date: 2002-11-13 pages: extension: .txt txt: ./txt/cord-007480-grndfx7b.txt cache: ./cache/cord-007480-grndfx7b.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-007480-grndfx7b.txt' === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97586 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === OMP: Error #34: System unable to allocate necessary resources for OMP thread: OMP: System error #11: Resource temporarily unavailable OMP: Hint Try decreasing the value of OMP_NUM_THREADS. /data-disk/reader-compute/reader-cord/bin/file2bib.sh: line 39: 97371 Aborted $FILE2BIB "$FILE" > "$OUTPUT" === file2bib.sh === id: cord-007495-gpz4gkv3 author: Weiss, M. title: Antibodies to berne virus in horses and other animals date: 2002-11-13 pages: extension: .txt txt: ./txt/cord-007495-gpz4gkv3.txt cache: ./cache/cord-007495-gpz4gkv3.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-007495-gpz4gkv3.txt' === file2bib.sh === id: cord-281081-rifr5uub author: Deng, Junhua title: Serological survey of SARS‐CoV‐2 for experimental, domestic, companion and wild animals excludes intermediate hosts of 35 different species of animals date: 2020-05-07 pages: extension: .txt txt: ./txt/cord-281081-rifr5uub.txt cache: ./cache/cord-281081-rifr5uub.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-281081-rifr5uub.txt' === file2bib.sh === id: cord-007497-nn5l5rai author: Garcia-Sanchez, J. title: Survey of rotavirus infection in a dairy herd: comparison between polycrylamide gel electrophoresis and two commercial tests date: 2002-11-13 pages: extension: .txt txt: ./txt/cord-007497-nn5l5rai.txt cache: ./cache/cord-007497-nn5l5rai.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-007497-nn5l5rai.txt' === file2bib.sh === id: cord-003208-lwirkob3 author: Yan, Liping title: Novel protein chip for the detection of antibodies against infectious bronchitis virus date: 2018-09-17 pages: extension: .txt txt: ./txt/cord-003208-lwirkob3.txt cache: ./cache/cord-003208-lwirkob3.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-003208-lwirkob3.txt' === file2bib.sh === id: cord-004101-0r2g5p1i author: Wang, Yu title: Self-assembly into virus–like particles of the recombinant capsid protein of porcine circovirus type 3 and its application on antibodies detection date: 2020-01-07 pages: extension: .txt txt: ./txt/cord-004101-0r2g5p1i.txt cache: ./cache/cord-004101-0r2g5p1i.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-004101-0r2g5p1i.txt' === file2bib.sh === id: cord-003471-tr3ageky author: Gaikwad, Satish S. title: Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein date: 2019-01-31 pages: extension: .txt txt: ./txt/cord-003471-tr3ageky.txt cache: ./cache/cord-003471-tr3ageky.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-003471-tr3ageky.txt' === file2bib.sh === id: cord-000434-ff2zadol author: Zhao, Rongmao title: Identification of a Highly Conserved H1 Subtype-Specific Epitope with Diagnostic Potential in the Hemagglutinin Protein of Influenza A Virus date: 2011-08-19 pages: extension: .txt txt: ./txt/cord-000434-ff2zadol.txt cache: ./cache/cord-000434-ff2zadol.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-000434-ff2zadol.txt' === file2bib.sh === id: cord-009922-t1hoox6e author: Dearden, C. J. title: Direct detection of rhinoviruses by an enzyme‐linked immunosorbent assay date: 2005-12-07 pages: extension: .txt txt: ./txt/cord-009922-t1hoox6e.txt cache: ./cache/cord-009922-t1hoox6e.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-009922-t1hoox6e.txt' === file2bib.sh === id: cord-007644-7bsixsgd author: Chirnside, E.D. title: Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus date: 2000-04-04 pages: extension: .txt txt: ./txt/cord-007644-7bsixsgd.txt cache: ./cache/cord-007644-7bsixsgd.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-007644-7bsixsgd.txt' === file2bib.sh === id: cord-011212-ovjdzyxv author: Pan, Qing title: Development and application of a novel ELISA for detecting antibodies against group I fowl adenoviruses date: 2019-12-14 pages: extension: .txt txt: ./txt/cord-011212-ovjdzyxv.txt cache: ./cache/cord-011212-ovjdzyxv.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-011212-ovjdzyxv.txt' === file2bib.sh === id: cord-009877-3cyz6o9c author: Barclay, Wendy S. title: Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions date: 2005-12-07 pages: extension: .txt txt: ./txt/cord-009877-3cyz6o9c.txt cache: ./cache/cord-009877-3cyz6o9c.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-009877-3cyz6o9c.txt' === file2bib.sh === id: cord-010578-uib9h1lb author: Mawle, Alison C. title: Seroepidemiology of Chronic Fatigue Syndrome: A Case-Control Study date: 1995-12-17 pages: extension: .txt txt: ./txt/cord-010578-uib9h1lb.txt cache: ./cache/cord-010578-uib9h1lb.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-010578-uib9h1lb.txt' === file2bib.sh === id: cord-275793-k0uvqcmp author: Xia, Hongyan title: A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies date: 2010-04-18 pages: extension: .txt txt: ./txt/cord-275793-k0uvqcmp.txt cache: ./cache/cord-275793-k0uvqcmp.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-275793-k0uvqcmp.txt' === file2bib.sh === id: cord-254384-mwzz1db5 author: Lu, Guilan title: Large-scale seroprevalence analysis of human metapneumovirus and human respiratory syncytial virus infections in Beijing, China date: 2011-02-10 pages: extension: .txt txt: ./txt/cord-254384-mwzz1db5.txt cache: ./cache/cord-254384-mwzz1db5.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-254384-mwzz1db5.txt' === file2bib.sh === id: cord-262268-gm99cadh author: Wang, Jingqiang title: Assessment of Immunoreactive Synthetic Peptides from the Structural Proteins of Severe Acute Respiratory Syndrome Coronavirus date: 2003-12-01 pages: extension: .txt txt: ./txt/cord-262268-gm99cadh.txt cache: ./cache/cord-262268-gm99cadh.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-262268-gm99cadh.txt' === file2bib.sh === id: cord-251943-jzaeaxam author: Zhang, Jian‐San title: A serological survey on neutralizing antibody titer of SARS convalescent sera date: 2005-08-24 pages: extension: .txt txt: ./txt/cord-251943-jzaeaxam.txt cache: ./cache/cord-251943-jzaeaxam.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-251943-jzaeaxam.txt' === file2bib.sh === id: cord-001446-mpuovmeb author: Bratcher, Preston E. title: Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA date: 2014-11-03 pages: extension: .txt txt: ./txt/cord-001446-mpuovmeb.txt cache: ./cache/cord-001446-mpuovmeb.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-001446-mpuovmeb.txt' === file2bib.sh === id: cord-277735-a9gkath5 author: Leung, Danny Tze Ming title: Antibody Response of Patients with Severe Acute Respiratory Syndrome (SARS) Targets the Viral Nucleocapsid date: 2004-07-15 pages: extension: .txt txt: ./txt/cord-277735-a9gkath5.txt cache: ./cache/cord-277735-a9gkath5.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-277735-a9gkath5.txt' === file2bib.sh === id: cord-003656-7mzsaz7a author: Wium, Martha title: DNA Vaccines Against Mycoplasma Elicit Humoral Immune Responses in Ostriches date: 2019-05-14 pages: extension: .txt txt: ./txt/cord-003656-7mzsaz7a.txt cache: ./cache/cord-003656-7mzsaz7a.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-003656-7mzsaz7a.txt' === file2bib.sh === id: cord-255390-dvp0luxe author: Jones, R. D title: Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles date: 2000-04-10 pages: extension: .txt txt: ./txt/cord-255390-dvp0luxe.txt cache: ./cache/cord-255390-dvp0luxe.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-255390-dvp0luxe.txt' === file2bib.sh === id: cord-011840-neowfhwg author: Liu, Weixiao title: Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops date: 2020-03-05 pages: extension: .txt txt: ./txt/cord-011840-neowfhwg.txt cache: ./cache/cord-011840-neowfhwg.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-011840-neowfhwg.txt' === file2bib.sh === id: cord-013348-lsksys56 author: Goto, Keiko title: Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 date: 2020-09-19 pages: extension: .txt txt: ./txt/cord-013348-lsksys56.txt cache: ./cache/cord-013348-lsksys56.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-013348-lsksys56.txt' === file2bib.sh === id: cord-003315-r1wkx0ml author: Jacobs, Sophie title: Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ date: 2018-11-01 pages: extension: .txt txt: ./txt/cord-003315-r1wkx0ml.txt cache: ./cache/cord-003315-r1wkx0ml.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-003315-r1wkx0ml.txt' === file2bib.sh === id: cord-254318-w8wrn9lx author: Díez, José-María title: Currently available intravenous immunoglobulin contains antibodies reacting against severe acute respiratory syndrome coronavirus 2 antigens date: 2020-05-13 pages: extension: .txt txt: ./txt/cord-254318-w8wrn9lx.txt cache: ./cache/cord-254318-w8wrn9lx.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-254318-w8wrn9lx.txt' === file2bib.sh === id: cord-281393-96j70n2z author: Capai, L. title: Seroprevalence of SARS-CoV-2 IgG antibodies, in Corsica (France), April and June 2020. date: 2020-09-30 pages: extension: .txt txt: ./txt/cord-281393-96j70n2z.txt cache: ./cache/cord-281393-96j70n2z.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-281393-96j70n2z.txt' === file2bib.sh === id: cord-262762-mgegswvn author: Callebaut, P. title: Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea date: 1982-09-30 pages: extension: .txt txt: ./txt/cord-262762-mgegswvn.txt cache: ./cache/cord-262762-mgegswvn.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-262762-mgegswvn.txt' === file2bib.sh === id: cord-313517-5ipj2z86 author: Fung, Joshua title: Antigen Capture Enzyme-Linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans date: 2019-09-14 pages: extension: .txt txt: ./txt/cord-313517-5ipj2z86.txt cache: ./cache/cord-313517-5ipj2z86.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-313517-5ipj2z86.txt' === file2bib.sh === id: cord-276989-441aclcc author: Liu, Jianbo title: Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum date: 2020-03-12 pages: extension: .txt txt: ./txt/cord-276989-441aclcc.txt cache: ./cache/cord-276989-441aclcc.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-276989-441aclcc.txt' === file2bib.sh === id: cord-003623-n01rgqyv author: Schuh, Amy J. title: Comparative analysis of serologic cross-reactivity using convalescent sera from filovirus-experimentally infected fruit bats date: 2019-04-30 pages: extension: .txt txt: ./txt/cord-003623-n01rgqyv.txt cache: ./cache/cord-003623-n01rgqyv.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-003623-n01rgqyv.txt' === file2bib.sh === id: cord-009784-kxa68zbc author: Bolton, Jessica S. title: Comparison of ELISA with electro-chemiluminescence technology for the qualitative and quantitative assessment of serological responses to vaccination date: 2020-04-17 pages: extension: .txt txt: ./txt/cord-009784-kxa68zbc.txt cache: ./cache/cord-009784-kxa68zbc.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-009784-kxa68zbc.txt' === file2bib.sh === id: cord-003859-k8wfyj9b author: Paweska, Janusz T. title: Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera date: 2019-07-24 pages: extension: .txt txt: ./txt/cord-003859-k8wfyj9b.txt cache: ./cache/cord-003859-k8wfyj9b.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-003859-k8wfyj9b.txt' === file2bib.sh === id: cord-012891-heqsfzkm author: Blanco Vázquez, Cristina title: Detection of latent forms of Mycobacterium avium subsp. paratuberculosis infection using host biomarker-based ELISAs greatly improves paratuberculosis diagnostic sensitivity date: 2020-09-03 pages: extension: .txt txt: ./txt/cord-012891-heqsfzkm.txt cache: ./cache/cord-012891-heqsfzkm.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-012891-heqsfzkm.txt' === file2bib.sh === id: cord-333026-9f6ecg30 author: Kompanikova, J. title: Microbiologic Methods in the Diagnostics of Upper Respiratory Tract Pathogens date: 2017-03-03 pages: extension: .txt txt: ./txt/cord-333026-9f6ecg30.txt cache: ./cache/cord-333026-9f6ecg30.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-333026-9f6ecg30.txt' === file2bib.sh === id: cord-301313-9595vm0k author: OKBA, NISREEN M.A. title: SARS-CoV-2 specific antibody responses in COVID-19 patients date: 2020-03-20 pages: extension: .txt txt: ./txt/cord-301313-9595vm0k.txt cache: ./cache/cord-301313-9595vm0k.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-301313-9595vm0k.txt' === file2bib.sh === id: cord-336899-zngp67tb author: Kadkhoda, Kamran title: COVID‐19: are neutralizing antibodies neutralizing enough? date: 2020-06-03 pages: extension: .txt txt: ./txt/cord-336899-zngp67tb.txt cache: ./cache/cord-336899-zngp67tb.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-336899-zngp67tb.txt' === file2bib.sh === id: cord-026559-xx52u01h author: Tripathi, Siddhartha title: Blood Plasma Microfluidic Device: Aiming for the Detection of COVID-19 Antibodies Using an On-Chip ELISA Platform date: 2020-06-10 pages: extension: .txt txt: ./txt/cord-026559-xx52u01h.txt cache: ./cache/cord-026559-xx52u01h.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-026559-xx52u01h.txt' === file2bib.sh === id: cord-317123-0tdfvlqd author: Tan, Xiaotian title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples date: 2020-04-21 pages: extension: .txt txt: ./txt/cord-317123-0tdfvlqd.txt cache: ./cache/cord-317123-0tdfvlqd.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-317123-0tdfvlqd.txt' === file2bib.sh === id: cord-334896-3g75spkc author: MINAMI, Shohei title: Establishment of serological test to detect antibody against ferret coronavirus date: 2016-03-03 pages: extension: .txt txt: ./txt/cord-334896-3g75spkc.txt cache: ./cache/cord-334896-3g75spkc.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-334896-3g75spkc.txt' === file2bib.sh === id: cord-312456-6lxc2rj2 author: Soltan, Mohamed A. title: Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species date: 2016-05-11 pages: extension: .txt txt: ./txt/cord-312456-6lxc2rj2.txt cache: ./cache/cord-312456-6lxc2rj2.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-312456-6lxc2rj2.txt' === file2bib.sh === id: cord-321691-46la29tm author: Hsueh, Po-Ren title: SARS Antibody Test for Serosurveillance date: 2004-09-17 pages: extension: .txt txt: ./txt/cord-321691-46la29tm.txt cache: ./cache/cord-321691-46la29tm.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-321691-46la29tm.txt' === file2bib.sh === id: cord-335591-r0x8yaqj author: Ohnishi, Kazuo title: Establishment and Characterization of Monoclonal Antibodies Against SARS Coronavirus date: 2007-11-28 pages: extension: .txt txt: ./txt/cord-335591-r0x8yaqj.txt cache: ./cache/cord-335591-r0x8yaqj.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-335591-r0x8yaqj.txt' === file2bib.sh === id: cord-349744-8cg5yj20 author: Lassaunière, Ria title: Evaluation of nine commercial SARS-CoV-2 immunoassays date: 2020-04-10 pages: extension: .txt txt: ./txt/cord-349744-8cg5yj20.txt cache: ./cache/cord-349744-8cg5yj20.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-349744-8cg5yj20.txt' === file2bib.sh === id: cord-305516-s1lvhknm author: Watanabe, Shumpei title: Epizootology and experimental infection of Yokose virus in bats date: 2008-09-11 pages: extension: .txt txt: ./txt/cord-305516-s1lvhknm.txt cache: ./cache/cord-305516-s1lvhknm.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-305516-s1lvhknm.txt' === file2bib.sh === id: cord-331277-fjsuo3yy author: Hoste, Alexis C.R. title: Two serological approaches for detection of antibodies to SARS-CoV-2 in different scenarios: A screening tool and a point-of-care test date: 2020-08-11 pages: extension: .txt txt: ./txt/cord-331277-fjsuo3yy.txt cache: ./cache/cord-331277-fjsuo3yy.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-331277-fjsuo3yy.txt' === file2bib.sh === id: cord-320559-up1q3k6q author: Dortmans, J.C.F.M. title: Porcine epidemic diarrhea virus (PEDV) introduction into a naive Dutch pig population in 2014 date: 2018-05-24 pages: extension: .txt txt: ./txt/cord-320559-up1q3k6q.txt cache: ./cache/cord-320559-up1q3k6q.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-320559-up1q3k6q.txt' === file2bib.sh === id: cord-322529-3xn5v54s author: Rodák, L. title: Verification of Sensitivity and Specificity of Group A Rotavirus Detection in Piglets Faeces with Monoclonal Blocking ELISA Methods date: 2004-06-30 pages: extension: .txt txt: ./txt/cord-322529-3xn5v54s.txt cache: ./cache/cord-322529-3xn5v54s.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-322529-3xn5v54s.txt' === file2bib.sh === id: cord-313193-q5zeoqlb author: Carrat, F. title: Seroprevalence of SARS-CoV-2 among adults in three regions of France following the lockdown and associated risk factors: a multicohort study. date: 2020-09-18 pages: extension: .txt txt: ./txt/cord-313193-q5zeoqlb.txt cache: ./cache/cord-313193-q5zeoqlb.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-313193-q5zeoqlb.txt' === file2bib.sh === id: cord-344581-h7ikjgic author: Ong, David S.Y. title: Comparison of diagnostic accuracies of rapid serological tests and ELISA to molecular diagnostics in patients with suspected COVID-19 presenting to the hospital date: 2020-06-02 pages: extension: .txt txt: ./txt/cord-344581-h7ikjgic.txt cache: ./cache/cord-344581-h7ikjgic.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-344581-h7ikjgic.txt' === file2bib.sh === id: cord-335121-ro3x3qa3 author: Ingram, George A. title: A comparative assessment of four serological methods used in the detection and measurement of anti-parasite antibodies in the serum of the amphibian, bufo viridis date: 1988-04-30 pages: extension: .txt txt: ./txt/cord-335121-ro3x3qa3.txt cache: ./cache/cord-335121-ro3x3qa3.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-335121-ro3x3qa3.txt' === file2bib.sh === id: cord-317223-juw4xt8q author: Pedersen, Niels C. title: The causes of false-positives encountered during the screening of old-world primates for antibodies to human and simian retroviruses by ELISA date: 1986-11-30 pages: extension: .txt txt: ./txt/cord-317223-juw4xt8q.txt cache: ./cache/cord-317223-juw4xt8q.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-317223-juw4xt8q.txt' === file2bib.sh === id: cord-318266-i8x80knq author: Böse, Reinhard title: Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot date: 1994-05-31 pages: extension: .txt txt: ./txt/cord-318266-i8x80knq.txt cache: ./cache/cord-318266-i8x80knq.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-318266-i8x80knq.txt' === file2bib.sh === id: cord-348660-qnbgywgy author: Yilmaz, Huseyin title: Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen date: 2018-07-09 pages: extension: .txt txt: ./txt/cord-348660-qnbgywgy.txt cache: ./cache/cord-348660-qnbgywgy.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-348660-qnbgywgy.txt' === file2bib.sh === id: cord-347374-mryazbnq author: Okba, Nisreen M.A. title: Severe Acute Respiratory Syndrome Coronavirus 2−Specific Antibody Responses in Coronavirus Disease Patients date: 2020-07-17 pages: extension: .txt txt: ./txt/cord-347374-mryazbnq.txt cache: ./cache/cord-347374-mryazbnq.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-347374-mryazbnq.txt' === file2bib.sh === id: cord-334277-g3go3u02 author: Kovac, Marc title: EDTA-Anticoagulated Whole Blood for SARS-CoV-2 Antibody Testing by Electrochemiluminescence Immunoassay (ECLIA) and Enzyme-Linked Immunosorbent Assay (ELISA) date: 2020-08-14 pages: extension: .txt txt: ./txt/cord-334277-g3go3u02.txt cache: ./cache/cord-334277-g3go3u02.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-334277-g3go3u02.txt' === file2bib.sh === id: cord-351952-lhhjax3s author: Pickering, Suzanne title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date: 2020-09-24 pages: extension: .txt txt: ./txt/cord-351952-lhhjax3s.txt cache: ./cache/cord-351952-lhhjax3s.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-351952-lhhjax3s.txt' === file2bib.sh === id: cord-301175-6alsigxk author: Okda, Faten title: Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus date: 2015-08-01 pages: extension: .txt txt: ./txt/cord-301175-6alsigxk.txt cache: ./cache/cord-301175-6alsigxk.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-301175-6alsigxk.txt' === file2bib.sh === id: cord-334165-7gfk554m author: Stadlbauer, Daniel title: SARS‐CoV‐2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup date: 2020-04-17 pages: extension: .txt txt: ./txt/cord-334165-7gfk554m.txt cache: ./cache/cord-334165-7gfk554m.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-334165-7gfk554m.txt' === file2bib.sh === id: cord-353614-z4fpy607 author: Kusano, Nario title: Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies date: 2007-02-16 pages: extension: .txt txt: ./txt/cord-353614-z4fpy607.txt cache: ./cache/cord-353614-z4fpy607.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-353614-z4fpy607.txt' === file2bib.sh === id: cord-318444-sgm24q1i author: Walter, Justin D. title: Sybodies targeting the SARS-CoV-2 receptor-binding domain date: 2020-05-16 pages: extension: .txt txt: ./txt/cord-318444-sgm24q1i.txt cache: ./cache/cord-318444-sgm24q1i.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-318444-sgm24q1i.txt' === file2bib.sh === id: cord-317057-c2bwky6e author: Pickering, S. title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date: 2020-06-04 pages: extension: .txt txt: ./txt/cord-317057-c2bwky6e.txt cache: ./cache/cord-317057-c2bwky6e.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-317057-c2bwky6e.txt' === file2bib.sh === id: cord-327890-ocisq7e4 author: Pallett, Scott J C title: Point-of-care serological assays for delayed SARS-CoV-2 case identification among health-care workers in the UK: a prospective multicentre cohort study date: 2020-07-24 pages: extension: .txt txt: ./txt/cord-327890-ocisq7e4.txt cache: ./cache/cord-327890-ocisq7e4.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-327890-ocisq7e4.txt' === file2bib.sh === /data-disk/reader-compute/reader-cord/bin/file2bib.sh: fork: retry: No child processes id: cord-344309-6c2wttxg author: Lin, Huixing title: Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein date: 2018-08-20 pages: extension: .txt txt: ./txt/cord-344309-6c2wttxg.txt cache: ./cache/cord-344309-6c2wttxg.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-344309-6c2wttxg.txt' === file2bib.sh === id: cord-348161-757c51xw author: Petrosova, A. title: Development of a highly sensitive, field operable biosensor for serological studies of Ebola virus in central Africa date: 2007-03-26 pages: extension: .txt txt: ./txt/cord-348161-757c51xw.txt cache: ./cache/cord-348161-757c51xw.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-348161-757c51xw.txt' === file2bib.sh === id: cord-317026-9zgc6xrb author: Zhao, Shan title: Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus date: 2019-11-30 pages: extension: .txt txt: ./txt/cord-317026-9zgc6xrb.txt cache: ./cache/cord-317026-9zgc6xrb.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-317026-9zgc6xrb.txt' === file2bib.sh === id: cord-343185-lbmbp9ca author: Hansen, C. B. title: SARS-CoV-2 antibody responses determine disease severity in COVID-19 infected individuals date: 2020-07-29 pages: extension: .txt txt: ./txt/cord-343185-lbmbp9ca.txt cache: ./cache/cord-343185-lbmbp9ca.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-343185-lbmbp9ca.txt' === file2bib.sh === id: cord-340960-abanr641 author: Brigger, D. title: Accuracy of serological testing for SARS‐CoV‐2 antibodies: first results of a large mixed‐method evaluation study date: 2020-09-30 pages: extension: .txt txt: ./txt/cord-340960-abanr641.txt cache: ./cache/cord-340960-abanr641.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 6 resourceName b'cord-340960-abanr641.txt' === file2bib.sh === id: cord-346320-ysgz6adr author: Arabi, Yaseen M. title: Feasibility of Using Convalescent Plasma Immunotherapy for MERS-CoV Infection, Saudi Arabia date: 2016-09-17 pages: extension: .txt txt: ./txt/cord-346320-ysgz6adr.txt cache: ./cache/cord-346320-ysgz6adr.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-346320-ysgz6adr.txt' === file2bib.sh === id: cord-346574-u28y1ttw author: Chen, Keyan title: Development and evaluation of an immunochromatographic strip for rapid detection of porcine hemagglutinating encephalomyelitis virus date: 2012-08-24 pages: extension: .txt txt: ./txt/cord-346574-u28y1ttw.txt cache: ./cache/cord-346574-u28y1ttw.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-346574-u28y1ttw.txt' === file2bib.sh === id: cord-281760-34wuttqw author: Pereira, E.P.V. title: Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review date: 2019-05-22 pages: extension: .txt txt: ./txt/cord-281760-34wuttqw.txt cache: ./cache/cord-281760-34wuttqw.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-281760-34wuttqw.txt' === file2bib.sh === id: cord-326232-668f53qc author: Toftaker, Ingrid title: Evaluation of a multiplex immunoassay for bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk against two indirect ELISAs using latent class analysis date: 2018-06-01 pages: extension: .txt txt: ./txt/cord-326232-668f53qc.txt cache: ./cache/cord-326232-668f53qc.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 2 resourceName b'cord-326232-668f53qc.txt' === file2bib.sh === id: cord-334968-gonx5taq author: Pignatelli, Jaime title: Lineage specific antigenic differences in porcine torovirus hemagglutinin-esterase (PToV-HE) protein date: 2013-12-23 pages: extension: .txt txt: ./txt/cord-334968-gonx5taq.txt cache: ./cache/cord-334968-gonx5taq.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-334968-gonx5taq.txt' === file2bib.sh === id: cord-342024-kaku49xd author: Espejo, Andrea P title: Review of Current Advances in Serologic Testing for COVID-19 date: 2020-06-25 pages: extension: .txt txt: ./txt/cord-342024-kaku49xd.txt cache: ./cache/cord-342024-kaku49xd.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-342024-kaku49xd.txt' === file2bib.sh === id: cord-344871-486sk4wc author: Wu, Jianping title: Biochemical and structural characterization of the interface mediating interaction between the influenza A virus non-structural protein-1 and a monoclonal antibody date: 2016-09-16 pages: extension: .txt txt: ./txt/cord-344871-486sk4wc.txt cache: ./cache/cord-344871-486sk4wc.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-344871-486sk4wc.txt' === file2bib.sh === id: cord-351498-bmq6zcb0 author: Martínez-Sernández, Victoria title: Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis date: 2018-03-21 pages: extension: .txt txt: ./txt/cord-351498-bmq6zcb0.txt cache: ./cache/cord-351498-bmq6zcb0.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-351498-bmq6zcb0.txt' === file2bib.sh === id: cord-022310-yc6xtw0s author: Lappin, Michael R. title: Microbiology and Infectious Disease date: 2011-12-15 pages: extension: .txt txt: ./txt/cord-022310-yc6xtw0s.txt cache: ./cache/cord-022310-yc6xtw0s.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-022310-yc6xtw0s.txt' === file2bib.sh === id: cord-311349-145kwny3 author: Mariani, Stefano title: Surface plasmon resonance applications in clinical analysis date: 2014-02-25 pages: extension: .txt txt: ./txt/cord-311349-145kwny3.txt cache: ./cache/cord-311349-145kwny3.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 5 resourceName b'cord-311349-145kwny3.txt' === file2bib.sh === id: cord-279406-wwdqh9qs author: Guzman, Norberto A. title: A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy date: 2020-07-30 pages: extension: .txt txt: ./txt/cord-279406-wwdqh9qs.txt cache: ./cache/cord-279406-wwdqh9qs.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 4 resourceName b'cord-279406-wwdqh9qs.txt' === file2bib.sh === id: cord-022353-q2k2krnm author: W. Quimby, Fred title: Clinical Chemistry of the Laboratory Mouse date: 2007-09-02 pages: extension: .txt txt: ./txt/cord-022353-q2k2krnm.txt cache: ./cache/cord-022353-q2k2krnm.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 3 resourceName b'cord-022353-q2k2krnm.txt' === file2bib.sh === id: cord-006828-i88on326 author: nan title: Abstracts DGRh-Kongress 2013 date: 2013-09-15 pages: extension: .txt txt: ./txt/cord-006828-i88on326.txt cache: ./cache/cord-006828-i88on326.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 6 resourceName b'cord-006828-i88on326.txt' === file2bib.sh === id: cord-014462-11ggaqf1 author: nan title: Abstracts of the Papers Presented in the XIX National Conference of Indian Virological Society, “Recent Trends in Viral Disease Problems and Management”, on 18–20 March, 2010, at S.V. University, Tirupati, Andhra Pradesh date: 2011-04-21 pages: extension: .txt txt: ./txt/cord-014462-11ggaqf1.txt cache: ./cache/cord-014462-11ggaqf1.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 5 resourceName b'cord-014462-11ggaqf1.txt' === file2bib.sh === id: cord-353190-7qcoxl81 author: Nicklas, Werner title: Viral Infections of Laboratory Mice date: 2012-05-17 pages: extension: .txt txt: ./txt/cord-353190-7qcoxl81.txt cache: ./cache/cord-353190-7qcoxl81.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 5 resourceName b'cord-353190-7qcoxl81.txt' === file2bib.sh === id: cord-022653-qa1uph35 author: nan title: Poster Discussion Session PDS date: 2017-08-30 pages: extension: .txt txt: ./txt/cord-022653-qa1uph35.txt cache: ./cache/cord-022653-qa1uph35.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 7 resourceName b'cord-022653-qa1uph35.txt' === file2bib.sh === id: cord-015147-h0o0yqv8 author: nan title: Oral Communications and Posters date: 2014-09-12 pages: extension: .txt txt: ./txt/cord-015147-h0o0yqv8.txt cache: ./cache/cord-015147-h0o0yqv8.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 5 resourceName b'cord-015147-h0o0yqv8.txt' === file2bib.sh === id: cord-006860-a3b8hyyr author: nan title: 40th Annual Meeting of the GTH (Gesellschaft für Thrombose- und Hämostaseforschung) date: 1996 pages: extension: .txt txt: ./txt/cord-006860-a3b8hyyr.txt cache: ./cache/cord-006860-a3b8hyyr.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 9 resourceName b'cord-006860-a3b8hyyr.txt' === file2bib.sh === id: cord-019347-tj3ye1mx author: nan title: ABSTRACT BOOK date: 2010-02-19 pages: extension: .txt txt: ./txt/cord-019347-tj3ye1mx.txt cache: ./cache/cord-019347-tj3ye1mx.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 7 resourceName b'cord-019347-tj3ye1mx.txt' === file2bib.sh === id: cord-022650-phsr10jp author: nan title: Abstracts TPS date: 2018-08-14 pages: extension: .txt txt: ./txt/cord-022650-phsr10jp.txt cache: ./cache/cord-022650-phsr10jp.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 20 resourceName b'cord-022650-phsr10jp.txt' === file2bib.sh === id: cord-023095-4dannjjm author: nan title: Research Abstract Program of the 2011 ACVIM Forum Denver, Colorado, June 15–18, 2011 date: 2011-05-03 pages: extension: .txt txt: ./txt/cord-023095-4dannjjm.txt cache: ./cache/cord-023095-4dannjjm.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 9 resourceName b'cord-023095-4dannjjm.txt' === file2bib.sh === id: cord-022501-9wnmdvg5 author: nan title: P1460 – P1884 date: 2015-12-28 pages: extension: .txt txt: ./txt/cord-022501-9wnmdvg5.txt cache: ./cache/cord-022501-9wnmdvg5.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 11 resourceName b'cord-022501-9wnmdvg5.txt' === file2bib.sh === id: cord-015021-pol2qm74 author: nan title: Third International Congress on the Immune Consequences of Trauma, Shock and Sepsis —Mechanisms and Therapeutic Approaches date: 1994 pages: extension: .txt txt: ./txt/cord-015021-pol2qm74.txt cache: ./cache/cord-015021-pol2qm74.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 22 resourceName b'cord-015021-pol2qm74.txt' === file2bib.sh === id: cord-001521-l36f1gp7 author: nan title: Oral and Poster Manuscripts date: 2011-04-08 pages: extension: .txt txt: ./txt/cord-001521-l36f1gp7.txt cache: ./cache/cord-001521-l36f1gp7.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 27 resourceName b'cord-001521-l36f1gp7.txt' === file2bib.sh === id: cord-022888-dnsdg04n author: nan title: Poster Sessions date: 2009-08-19 pages: extension: .txt txt: ./txt/cord-022888-dnsdg04n.txt cache: ./cache/cord-022888-dnsdg04n.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 24 resourceName b'cord-022888-dnsdg04n.txt' === file2bib.sh === id: cord-031907-ilhr3iu5 author: nan title: ISEV2020 Abstract Book date: 2020-07-15 pages: extension: .txt txt: ./txt/cord-031907-ilhr3iu5.txt cache: ./cache/cord-031907-ilhr3iu5.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 26 resourceName b'cord-031907-ilhr3iu5.txt' === file2bib.sh === id: cord-022940-atbjwpo5 author: nan title: Poster Sessions date: 2016-09-07 pages: extension: .txt txt: ./txt/cord-022940-atbjwpo5.txt cache: ./cache/cord-022940-atbjwpo5.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 17 resourceName b'cord-022940-atbjwpo5.txt' === file2bib.sh === id: cord-015394-uj7fe5y6 author: nan title: Scientific Abstracts date: 2008-12-23 pages: extension: .txt txt: ./txt/cord-015394-uj7fe5y6.txt cache: ./cache/cord-015394-uj7fe5y6.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 X-Parsed-By ['org.apache.tika.parser.DefaultParser', 'org.apache.tika.parser.csv.TextAndCSVParser'] X-TIKA:content_handler ToTextContentHandler X-TIKA:embedded_depth 0 X-TIKA:parse_time_millis 16 resourceName b'cord-015394-uj7fe5y6.txt' Que is empty; done keyword-elisa-cord === reduce.pl bib === id = cord-000434-ff2zadol author = Zhao, Rongmao title = Identification of a Highly Conserved H1 Subtype-Specific Epitope with Diagnostic Potential in the Hemagglutinin Protein of Influenza A Virus date = 2011-08-19 pages = extension = .txt mime = text/plain words = 5752 sentences = 307 flesch = 51 summary = The highly conserved H1 subtype-specific immunodominant epitope may form the basis for developing novel assays for sero-diagnosis and active surveillance against H1N1 IAVs. Influenza A viruses (IAVs), members of the Orthomyxoviridae family, are highly contagious to a variety of avian and mammalian species. To confirm that these antibodies can recognize the HA antigen, the reactivity of the anti-peptide sera were evaluated by Western blot and ELISA against the purified HA0 protein of H1N1pdm virus. The sensitivity and specificity of peptide-ELISA versus HI test was 96.5% and 74.4%, respectively, indicating the potential of the peptide-ELISA method in detecting antibody against H1-subtype IAVs. In the present study, we identified immunodominant linear B cell epitopes on the H1N1pdm virus HA protein by a peptide scanning approach using H1N1pdm patients sera. To screen the H1-subtype specific epitopes, a set of 50 peptides spanning the amino acid sequences of the HA protein ectodomain of pandemic A/H1N1 2009 (H1N1pdm) influenza virus strain A/ California/04/2009 were synthesized. cache = ./cache/cord-000434-ff2zadol.txt txt = ./txt/cord-000434-ff2zadol.txt === reduce.pl bib === id = cord-003208-lwirkob3 author = Yan, Liping title = Novel protein chip for the detection of antibodies against infectious bronchitis virus date = 2018-09-17 pages = extension = .txt mime = text/plain words = 3973 sentences = 220 flesch = 52 summary = RESULTS: We have developed two indirect microarray methods to detect antibodies against IBV: a chemiluminescent immunoassay test (CIT) and a rapid diagnostic test (RDT). Compared with these methods, enzyme-linked immunosorbent assay (ELISA) has been widely used for testing IBV early infection and continuous infection, and this technique can be used for both antigenic and antibody detection. The data showed that 130 serum samples were positive for antibodies against IBV, and 14 samples were negative, similar to the results of the IDEXX IBV Ab Test kit with the nsp5 concentration of 0.2 mg/mL (Table 4 ). The specific experiments of the RDT showed that no cross-reaction Fig. 4 a Distribution of the SNRs of the IDEXX-positive (n = 142) and IDEXX-negative (n = 42) serum samples of the clinical sera obtained from the IBV protein microarray. cache = ./cache/cord-003208-lwirkob3.txt txt = ./txt/cord-003208-lwirkob3.txt === reduce.pl bib === id = cord-004101-0r2g5p1i author = Wang, Yu title = Self-assembly into virus–like particles of the recombinant capsid protein of porcine circovirus type 3 and its application on antibodies detection date = 2020-01-07 pages = extension = .txt mime = text/plain words = 4241 sentences = 232 flesch = 58 summary = The purified eCap and dCap were identified by transmission electron microscopy (TEM), a large number of round hollow particles with a diameter of 10 nm virus-like particles (VLPs) were observed in eCap, whereas irregular aggregation of proteins observed in dCap. After formation the VLPs were applied as a coating antigen to establish an indirect ELISA (I-ELISA) for detection of PCV3-specific antibody in swine serum. To the best of our knowledge, this is the first report of self-assembly into VLPs of PCV3 recombinant Cap. Our results demonstrated that the VLP-based I-ELISA will be a valuable tool for detecting the presence of PCV3 antibodies in serum samples and will facilitate screening of large numbers of swine serum for clinical purposes. Generation in yeast of recombinant virus-like particles of porcine circovirus type 2 capsid protein and their use for a serologic assay and development of monoclonal antibodies cache = ./cache/cord-004101-0r2g5p1i.txt txt = ./txt/cord-004101-0r2g5p1i.txt === reduce.pl bib === id = cord-003471-tr3ageky author = Gaikwad, Satish S. title = Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein date = 2019-01-31 pages = extension = .txt mime = text/plain words = 3398 sentences = 199 flesch = 51 summary = PURPOSE: The aim of the present study was to develop a serodiagnostic test for differentiation infected from vaccinated animal (DIVA) strategy accompanying the marker vaccine lacking an immunodominant epitope (IDE) of nucleoprotein of Newcastle disease virus (NDV). Epitope repeat protein (ERP) gene encoding eight repeats of an IDE sequence containing ETQFLDLMRAVANSMR (C-terminal IDE aa 444-459) on NDV NP separated by tetra-glycine linker was synthesized commercially using codon optimization for baculovirus expression. In addition to SPF chicken serum, different hyper-immune chicken sera to NDV, H9N2 avian influenza virus (AIV), infectious bronchitis virus (IBV), and infectious bursal disease virus (IBDV) were tested for specificity of rERP using an indirect ELISA assay. Second and third groups were sera taken 14 dpi from SPF chickens vaccinated with IDE deleted recombinant Newcastle disease virus (rNDV) [4] and NDV LaSota strain [19] , respectively, which kindly supplied by the OIE reference laboratory for ND, Korea. cache = ./cache/cord-003471-tr3ageky.txt txt = ./txt/cord-003471-tr3ageky.txt === reduce.pl bib === id = cord-007480-grndfx7b author = Koopmans, M. title = Seroepidemiology of Breda virus in cattle using ELISA date = 2002-11-13 pages = extension = .txt mime = text/plain words = 3354 sentences = 176 flesch = 55 summary = Two direct blocking enzyme linked immunosorbent assays (ELISA) for the detection of antibodies to Breda virus in sera of cattle were compared. This "Breda virus" (BRV) is morphologically and antigenically different from known bovine viruses and causes diarrhea in gnotobiotic and colostrum-deprived calves (Woode et al., 1982) . In the first approach (consecutive method) a 1:400 dilution of the BRV2 antigen preparation, purified from calf feces was added to the coated wells in ELISA buffer 1 (PBS supplemented with 0.35 M NaC1, 1 mM EDTA pH 7.5 and 0.05% Tween 80 ). When using the consecutive method more animals were found negative for antibodies to BRV2 or had low blocking percentages (Table 1 ) . In Fig. 3 , the percentages of animals with antibodies to BRV ( >~ 20% blocking, corresponding to a serum titer of/> 30 ) are shown for different age groups (n--244). cache = ./cache/cord-007480-grndfx7b.txt txt = ./txt/cord-007480-grndfx7b.txt === reduce.pl bib === id = cord-003315-r1wkx0ml author = Jacobs, Sophie title = Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ date = 2018-11-01 pages = extension = .txt mime = text/plain words = 6190 sentences = 337 flesch = 59 summary = We next developed a firefly luciferase-based reporter cell line, named Fawa-λ-luc, to detect IFN-λ in biological fluids with high specificity and sensitivity. This bioassay was as sensitive as a commercially available enzyme-linked immunosorbent assay in detecting mouse IFN-λ in cell culture supernatant, as well as in serum and bronchoalveolar lavage samples of virus-infected mice. Unlike mouse and human type I IFNs that exhibit strongly species-specific activity (Veomett and Veomett 1979) , type III IFNs were reported to act on cell types from both origins (Lasfar and others 2006; Hermant and others 2014) . IFNl4 was quantified by a cytopathic effect reduction assay in A549 cells, using recombinant human IFN-l3 as a standard, which was reported to have a similar specific activity (Hamming and others 2013). We then compared the sensitivities of the Fawa-l-luc and of the ELISA assays to detect IFN-l in mouse serum and BAL samples. cache = ./cache/cord-003315-r1wkx0ml.txt txt = ./txt/cord-003315-r1wkx0ml.txt === reduce.pl bib === id = cord-003656-7mzsaz7a author = Wium, Martha title = DNA Vaccines Against Mycoplasma Elicit Humoral Immune Responses in Ostriches date = 2019-05-14 pages = extension = .txt mime = text/plain words = 5575 sentences = 308 flesch = 53 summary = Currently there are no commercially available vaccines against ostrich-infecting mycoplasmas and this study therefore set out to develop and evaluate the use of a DNA vaccine against mycoplasma infections in ostriches using an OppA protein as antigen. The ability of the DNA vaccines to elicit an anti-OppA antibody response was evaluated by ELISA using the recombinant OppA protein of Ms03 as coating antigen. In this study we report, for the first time, that a DNA vaccine can elicit a humoral immune response in ostriches using OppA as antigen. The controls were serum samples representing the week 0, 3, 6, and 9 sampling points of a single ostrich, randomly selected from the pCI-neo_oppA 1,200 µg group based on high titers produced after vaccination. In this study, DNA vaccines were developed for ostriches using the oppA gene of an ostrich-infecting mycoplasma (Ms03) as vaccine antigen. cache = ./cache/cord-003656-7mzsaz7a.txt txt = ./txt/cord-003656-7mzsaz7a.txt === reduce.pl bib === id = cord-009922-t1hoox6e author = Dearden, C. J. title = Direct detection of rhinoviruses by an enzyme‐linked immunosorbent assay date = 2005-12-07 pages = extension = .txt mime = text/plain words = 4065 sentences = 205 flesch = 55 summary = This paper describes the first enzyme‐linked immunosorbent assay for the detection of rhinovirus antigens in clinical specimens (nasal washings), either directly or following overnight cell culture amplification. Secondly, a direct ELISA system was developed in which the nasal washings or control antigen (uninfected tissue culture fluid) were added directly to each of a set of duplicate ELISA plate wells coated with either pre-or postchallenge rabbit anti HRV-EL hyperimmune serum. Figure 2 shows the results obtained with nasal washings, collected from three volunteers on consecutive days following HKV-EL or saline challenge, tested in both the cell-culture-amplified (CCA)-ELISA and direct ELISA systems. Although we would like to emphasise that our data are preliminary, both the direct and CCA-ELISAs gave a good correlation with virus isolation when used to detect rhinovirus antigens in nasal washings (obtained from 18 volunteers challenged with either HRV-EL or saline). cache = ./cache/cord-009922-t1hoox6e.txt txt = ./txt/cord-009922-t1hoox6e.txt === reduce.pl bib === id = cord-007497-nn5l5rai author = Garcia-Sanchez, J. title = Survey of rotavirus infection in a dairy herd: comparison between polycrylamide gel electrophoresis and two commercial tests date = 2002-11-13 pages = extension = .txt mime = text/plain words = 3578 sentences = 165 flesch = 54 summary = Faecal samples taken from 15 cows before and after calving as well as faeces taken from their calves daily from birth to two weeks of life were tested for rotavirus by polyacrylamide gel electrophoresis (PAGE) and compared with an ELISA and a latex agglutination commercial test. Various methods have been developed for rapid detection of rotaviruses in faecal samples; these include electron microscopy (considered for a long time as the reference method), immune electron microscopy, counter immuno-electrophoresis, complement fixation, fluorescent antibody tests, different immunoassays using antibodies labelled with radioisotopes or enzymes (ELISA), latex agglutination and polyacrylamide gel electrophoresis (PAGE). All the faeces samples (120 from cows and 240 from calves) were tested for the presence of rotaviruses by polyacrylamide gel electrophoresis (PAGE), a commercial ELISA kit and a commercial latex agglutination kit. Another three discrepancies were detected in which the samples were positive by PAGE and negative by both ELISA and latex agglutination and all of them corresponded to two calves with a discontinuous shedding pattern (calf no. cache = ./cache/cord-007497-nn5l5rai.txt txt = ./txt/cord-007497-nn5l5rai.txt === reduce.pl bib === id = cord-011212-ovjdzyxv author = Pan, Qing title = Development and application of a novel ELISA for detecting antibodies against group I fowl adenoviruses date = 2019-12-14 pages = extension = .txt mime = text/plain words = 3884 sentences = 223 flesch = 47 summary = Since 2015, outbreaks of hepatitis-hydropericardium syndrome (HPS) caused by a novel genotype of fowl adenovirus 4 (FAdV-4) infection have created serious economic losses in China. In one recent study, recombinant fiber-based indirect ELISA was used to detect serum samples from chickens experimentally inoculated with different FAdV-1 or FAdV-4 strains (Feichtner et al. A recombinant hexon-based single serum dilution ELISA was also developed to measure the hexon-specific antibodies against FAdV-4 in sera of chickens (Rajasekhar and Roy 2014) . In this study, we developed a group-specific and sensitive ELISA based on the novel genotype of FAdV-4 for detecting antibodies against twelve FAdV-I serotypes. The common ELISA developed in our study was applied to detect serum samples from SPF chickens inoculated with inactivated FAdV-1, FAdV-4, and FAdV-8a, and showed high sensitivity for all three FAdV hypervirulent serotypes. Furthermore, the ELISA showed higher sensitivity in detecting serum samples of HPS caused by the novel FAdV-4 genotype that has recently emerged in China. cache = ./cache/cord-011212-ovjdzyxv.txt txt = ./txt/cord-011212-ovjdzyxv.txt === reduce.pl bib === id = cord-007495-gpz4gkv3 author = Weiss, M. title = Antibodies to berne virus in horses and other animals date = 2002-11-13 pages = extension = .txt mime = text/plain words = 2113 sentences = 103 flesch = 51 summary = Positive reactions were also obtained in serum neutralization tests and ELISA using small numbers of horse sera from Germany, France and the U.S.A. The results of neutralization tests and ELISA were correlated in 83% of random samples tested; 13% were neutralization-positive and ELISA-negative and in 4% the inverse was observed. The activity of Berne virus in the Swiss adult horse population was evalPositive reactions in serum neutralization of small numbers of randomlycollected equine sera from Germany (8/11) and the USA (24/38) and in ELISA of samples from Southern France (10/28) and the USA (12/16) were recorded. As shown in Table IV , high percentages of serum samples with elevated neutralization titers were encountered in all ungulates studied (horse, cattle, goat, sheep and pig). We have preliminary evidence (from neutralization tests using 31 randomly collected cattle sera) that a Berne-related virus is active also in the Dutch cattle population; in only 3 samples titers were <10 and in 10 sera values of >50 were recorded. cache = ./cache/cord-007495-gpz4gkv3.txt txt = ./txt/cord-007495-gpz4gkv3.txt === reduce.pl bib === id = cord-003623-n01rgqyv author = Schuh, Amy J. title = Comparative analysis of serologic cross-reactivity using convalescent sera from filovirus-experimentally infected fruit bats date = 2019-04-30 pages = extension = .txt mime = text/plain words = 6363 sentences = 277 flesch = 37 summary = To evaluate the ability of our system comprising seven filovirus-specific indirect ELISAs to predict the filovirus species most antigenically similar to the species responsible for past infection, we tested seven Marburg virus convalescent serum 35 or whole blood 36 samples collected from experimentally inoculated ERBs. Five of these samples www.nature.com/scientificreports www.nature.com/scientificreports/ were collected four weeks post primary Marburg virus inoculation 35, 36 , while two of the samples were collected at 23 and 27 weeks post primary inoculation following a "natural" boost (i.e., Marburg virus-specific antibody levels waned in these bats and then increased following contact with infectious cagemates) 36 . Although significant levels of serological IgG cross-reactivity were observed between the prime-boost filovirus-specific antisera and some of the filovirus antigens, when the overall covariance of the seven-individual indirect ELISAs in the system were considered, we were able to predict the filovirus species responsible for past infection 100% of the time using as little as 25 μL of sera (each serum was tested against each antigen in duplicate). cache = ./cache/cord-003623-n01rgqyv.txt txt = ./txt/cord-003623-n01rgqyv.txt === reduce.pl bib === id = cord-001446-mpuovmeb author = Bratcher, Preston E. title = Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA date = 2014-11-03 pages = extension = .txt mime = text/plain words = 4873 sentences = 226 flesch = 43 summary = Circulating levels of SP-D have been examined for their potential use as a biomarker in various diseases including dermatitis [2, 3] , acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] , periodontitis [14] , interstitial pulmonary fibrosis (IPF) [10, 12, [15] [16] [17] [18] [19] [20] [21] [22] [23] , chronic obstructive pulmonary disease (COPD) [15, [24] [25] [26] [27] [28] [29] [30] [31] [32] [33] [34] [35] [36] , emphysema [37] , cystic fibrosis (CF) [15, 38, 39] , coronary disease [40, 41] , sclerosis [42] [43] [44] [45] [46] , cancer [47, 48] , sarcoidosis [21, 49] , allergies [28, [50] [51] [52] , rheumatoid arthritis [53, 54] , and respiratory infections [18, [55] [56] [57] [58] [59] [60] . Serum levels of surfactant proteins A and D are useful biomarkers for interstitial lung disease in patients with progressive systemic sclerosis cache = ./cache/cord-001446-mpuovmeb.txt txt = ./txt/cord-001446-mpuovmeb.txt === reduce.pl bib === id = cord-003859-k8wfyj9b author = Paweska, Janusz T. title = Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera date = 2019-07-24 pages = extension = .txt mime = text/plain words = 6407 sentences = 308 flesch = 51 summary = title: Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera Diagnostic performance of three indirect enzyme-linked immunosorbent assays (I-ELISA) was evaluated for the detection of IgG antibody to Ebola virus (EBOV) in human sera. Validation data sets derived from individual sera collected in South Africa (SA), representing an EBOV non-endemic country, and from sera collected during an Ebola disease (EBOD) outbreak in Sierra Leone (SL), were categorized according to the compounded results of the three I-ELISAs and real time reverse-transcription polymerase chain reaction (RT-PCR). The purpose of this study was to evaluate and compare the diagnostic performance of EBOV IgG-indirect ELISAs based on antigens produced by classical virological and recombinant protein expression methods using human serum panels from EBOV non-infected and EBOV infected humans. cache = ./cache/cord-003859-k8wfyj9b.txt txt = ./txt/cord-003859-k8wfyj9b.txt === reduce.pl bib === id = cord-012891-heqsfzkm author = Blanco Vázquez, Cristina title = Detection of latent forms of Mycobacterium avium subsp. paratuberculosis infection using host biomarker-based ELISAs greatly improves paratuberculosis diagnostic sensitivity date = 2020-09-03 pages = extension = .txt mime = text/plain words = 7818 sentences = 354 flesch = 43 summary = The aim of this study was to evaluate the diagnostic potential of commercial ELISAs based on detection of these five bovine biomarkers to detect latent and patent forms of MAP infection in naturally infected cattle, using reference serum samples from well characterized animals with focal, multifocal and diffuse histological lesions in their intestinal or associated lymphoid tissues [40]. The diagnostic performance of the ABCA13, MMP8 and SPARC-based ELISAs for the focal and any type of lesion groups was compared with the IDEXX ELISA and additionally with other conventional PTB diagnosis methods such as specific fecal and tissue real-time PCR and bacteriological culture ( Table 4 ). Our results indicate that the ABCA13, SPARC and MMP8-based ELISAs have higher AUC values and sensitivities than the IDEXX ELISA and other current diagnostic methods for detection of animals with focal, multifocal and any type of histopathological lesions, respectively. cache = ./cache/cord-012891-heqsfzkm.txt txt = ./txt/cord-012891-heqsfzkm.txt === reduce.pl bib === id = cord-007644-7bsixsgd author = Chirnside, E.D. title = Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus date = 2000-04-04 pages = extension = .txt mime = text/plain words = 4054 sentences = 198 flesch = 49 summary = authors: Chirnside, E.D.; Francis, P.M.; De Vries, A.A.F.; Sinclaira, R.; Mumford, J.A. title: Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus A recombinant glutathione-S-transferase fusion protein expressing amino acids 55–98 of equine arteritis virus (EAV) G(L) (rG(L)55–98) was tested in an ELISA for its ability to detect serum antibodies to EAV. This paper describes an indirect ELISA using a recombinant glutathione-Stransferase fusion protein (Smith and Johnson, 1988) as an antigen to screen equine sera for the presence of antibodies to EAV, and its evaluation as a diagnostic test with large numbers of equine serum samples. By testing > 1500 equine sera in ELISA to G,55-98, we have demonstrated that amino acid residues 55-98 of the Bucyrus strain of EAV G,_ encompass a highly immunoreactive antigen which correlates closely with the host virus neutralizing response. cache = ./cache/cord-007644-7bsixsgd.txt txt = ./txt/cord-007644-7bsixsgd.txt === reduce.pl bib === id = cord-014462-11ggaqf1 author = nan title = Abstracts of the Papers Presented in the XIX National Conference of Indian Virological Society, “Recent Trends in Viral Disease Problems and Management”, on 18–20 March, 2010, at S.V. University, Tirupati, Andhra Pradesh date = 2011-04-21 pages = extension = .txt mime = text/plain words = 35453 sentences = 1711 flesch = 49 summary = Molecular diagnosis based on reverse transcription (RT)-PCR s.a. one step or nested PCR, nucleic acid sequence based amplification (NASBA), or real time RT-PCR, has gradually replaced the virus isolation method as the new standard for the detection of dengue virus in acute phase serum samples. Non-genetic methods of management of these diseases include quarantine measures, eradication of infected plants and weed hosts, crop rotation, use of certified virus-free seed or planting stock and use of pesticides to control insect vector populations implicated in transmission of viruses. The results of this study indicate that NS1 antigen based ELISA test can be an useful tool to detect the dengue virus infection in patients during the early acute phase of disease since appearance of IgM antibodies usually occur after fifth day of the infection. The studies showed high level of expression in case of constructed vector as compared to infected virus for the specific protein. cache = ./cache/cord-014462-11ggaqf1.txt txt = ./txt/cord-014462-11ggaqf1.txt === reduce.pl bib === id = cord-009877-3cyz6o9c author = Barclay, Wendy S. title = Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions date = 2005-12-07 pages = extension = .txt mime = text/plain words = 3078 sentences = 144 flesch = 47 summary = In a preliminary study involving 12 volunteers inoculated intranasally with human rhinovirus EL (HRV-EL), we showed that the presence of circulating rhinovirus-specific IgA, rather than IgG, protected volunteers from infection [Barclay and Al-Nakib, 19871. Figure 1 shows a) the mean HRV-2-neutralising antibody, b) specific IgG and c) IgA in the serum, and d) specific IgA in nasal secretions (normalized for total IgA) in pre-inoculation samples from the three groups of volunteers. Recently, a similar study involving samples obtained from volunteers inoculated with another rhinovirus serotype (HRV-9) and tested by ELISA for homologous antibodies, appears to confirm the association between ELISA-serumspecific IgA and protection against reinfection (Callow and Sergeant, unpublished data). The size of rises in HRV-2-specific IgA in the serum after virus inoculation showed a significant negative correlation with the concentration of specific IgA in the pre-inoculation nasal secretions and with the presence of serum neutralising antibody, indicating that volunteers who were most susceptible to infection and illness, in fact, showed the largest antibody response. cache = ./cache/cord-009877-3cyz6o9c.txt txt = ./txt/cord-009877-3cyz6o9c.txt === reduce.pl bib === id = cord-009784-kxa68zbc author = Bolton, Jessica S. title = Comparison of ELISA with electro-chemiluminescence technology for the qualitative and quantitative assessment of serological responses to vaccination date = 2020-04-17 pages = extension = .txt mime = text/plain words = 6166 sentences = 286 flesch = 47 summary = The PfCSP-FL protein is comprised of 26 Tyr -127 Asp linked to 207 Pro -383 Ser [4] ; "Repeat" is a Keywords: Serology, Vaccine, Antigen, Multiplex, Antigenic competition, ELISA, Electro-chemiluminescence 32-mer peptide representing the central Repeat region (NANP 8 ); C-term is a recombinant protein representing the C-terminal fragment (AA 207-383); Pf16 is an epitope within the C-terminus that has been used as a functional marker when evaluating anti-CSP antibodies induced by vaccination [4, 7, 8] . To establish a multiplex assay using an ECLIA platform, several parameters (i.e., antigen coating concentration, antigenic competition between closely related antigens, sample dilutions) were optimized and the performance of the assay determined in regards to specificity, linearity, and throughput. cache = ./cache/cord-009784-kxa68zbc.txt txt = ./txt/cord-009784-kxa68zbc.txt === reduce.pl bib === id = cord-011840-neowfhwg author = Liu, Weixiao title = Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops date = 2020-03-05 pages = extension = .txt mime = text/plain words = 3920 sentences = 232 flesch = 52 summary = title: Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops OBJECTIVES: To develop a sensitive monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) to detect Vip3Aa in genetically modified (GM) crops and their products. A sensitive monoclonal antibody-based sandwich ELISA was developed to detect Vip3Aa in GM crops and their products. These antibodies were then employed to develop a sandwich ELISA for sensitive, direct and convenient measurement of the Vip3Aa concentration in GM crops and their products. To the best of our knowledge, this is the first quantitative monoclonal antibody-based ELISA method reported for the sensitive detection of Vip3Aa proteins. In this study, we describe a sensitive monoclonal antibody-based ELISA method for the detection of Vip3Aa in GM crops and their products. Development of monoclonal antibodies against Cry1Ab protein from Bacillus thuringiensis and their application in an ELISA for detection of transgenic Bt-maize cache = ./cache/cord-011840-neowfhwg.txt txt = ./txt/cord-011840-neowfhwg.txt === reduce.pl bib === id = cord-281081-rifr5uub author = Deng, Junhua title = Serological survey of SARS‐CoV‐2 for experimental, domestic, companion and wild animals excludes intermediate hosts of 35 different species of animals date = 2020-05-07 pages = extension = .txt mime = text/plain words = 1515 sentences = 86 flesch = 55 summary = In this study, 1,914 serum samples from 35 animal species were used for detection of SARS‐CoV‐2‐specific antibodies using double‐antigen sandwich ELISA after validating its specificity and sensitivity. The results showed that no SARS‐CoV‐2‐specific antibodies were detected in above samples which excluded the possibility of 35 animal species as intermediate host for SARS‐CoV‐2. The results showed that no SARS-CoV-2-specific antibodies were detected in above species of animals including pangolin which has been reported as an intermediate host of SARS-CoV-2 (Kangpeng Xiao, 2020) . After confirming the specificity, sensitivity and suitability of SARS-CoV-2 ELISA kit for different species of experimental animals, clinical serum samples from domestic livestock (pig, cow, sheep, horse), poultry (chicken, duck, goose), experimental animal (mice, rat and rhesus monkey), companion animal (dog and cat) and wild animals (camel, fox, mink, alpaca, ferret, bamboo rat, peacock, eagle, tiger rhinoceros, pangolin, leopard cat, jackal, giant panda, masked civet, porcupine, bear, yellow-throated marten, weasel, red pandas and wild boar) were used for antibody detection. cache = ./cache/cord-281081-rifr5uub.txt txt = ./txt/cord-281081-rifr5uub.txt === reduce.pl bib === id = cord-013348-lsksys56 author = Goto, Keiko title = Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 date = 2020-09-19 pages = extension = .txt mime = text/plain words = 4742 sentences = 247 flesch = 50 summary = title: Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 From the resultant hybridoma clones, we selected nine clones producing mAbs reactive to the HPeV3-VP0 antigen, based on enzyme-linked immunosorbent assay (ELISA). Although RT-PCR-based diagnostic tests targeting 5 -UTR of the HPeV3 genome were developed for HPeV3 detection in clinical samples, there is currently no diagnostic method for detecting the viral antigens. To develop a rapid and effective diagnostic strategy, there is an urgent need to produce highly specific monoclonal antibodies (mAbs) toward HPeV3 antigens. As a result, we obtained nine mAb clones for characterization, and thereafter, generated an ELISA system that is specifically able to detect the HPeV3 VP0 antigens. We next performed antigen-capture ELISA with recombinant VP0 protein and virions released into the cell-culture supernatant of the HPeV3-infected cells. In this study, we sought to generate specific mAbs and develop an ELISA test for the detection of HPeV3 VP0 antigen. cache = ./cache/cord-013348-lsksys56.txt txt = ./txt/cord-013348-lsksys56.txt === reduce.pl bib === id = cord-255390-dvp0luxe author = Jones, R. D title = Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles date = 2000-04-10 pages = extension = .txt mime = text/plain words = 4834 sentences = 237 flesch = 39 summary = Abstract The purpose of this subacute 22-day study was to evaluate methods for canine circulating immunoglobulins (IgM, IgG, and IgE) and select Band T-lymphocyte populations (CD4-helpers, CD8-suppressors, pan-T and pan-B) for immunotoxicity testing using an organ system (concordance) approach. The flow cytometry method was acceptable for measuring select canine lymphocyte populations and detecting the expected decrease in B cells due to cyclophosphamide treatment. The purpose of this study was to evaluate immunoglobulins (IgM, IgG, IgE) by ELISA and lymphocytes (CD4-helpers, CD8-suppressors, pan-T CD5 and pan-B CD21) by flow cytometry for detecting adverse effects in dogs administered positive control challenges to the immune system. The next day following each immunization, serum samples were taken for analysis of IgG, IgM and IgE, along with clinical chemistry, hematology, urinalysis, and flow cytometry on day 8, to evaluate possible influences of repeated immunization. cache = ./cache/cord-255390-dvp0luxe.txt txt = ./txt/cord-255390-dvp0luxe.txt === reduce.pl bib === id = cord-010578-uib9h1lb author = Mawle, Alison C. title = Seroepidemiology of Chronic Fatigue Syndrome: A Case-Control Study date = 1995-12-17 pages = extension = .txt mime = text/plain words = 2570 sentences = 164 flesch = 49 summary = We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We conducted serological tests for a large number of infectious agents as part of a case-control study assessing risk factors for CFS. Antibodies against human T-lymphotrophic virus types I and II were detected with an ELISA, and confirmatory testing was performed by western blotting [5] . All other agents tested were detected in ;;:::25% of CFS cases, and antibody levels were compared between cases and controls. Evidence for active Epstein-Barr virus infection in patients with persistent unexplained illness: elevated anti-early antigen antibodies cache = ./cache/cord-010578-uib9h1lb.txt txt = ./txt/cord-010578-uib9h1lb.txt === reduce.pl bib === id = cord-277735-a9gkath5 author = Leung, Danny Tze Ming title = Antibody Response of Patients with Severe Acute Respiratory Syndrome (SARS) Targets the Viral Nucleocapsid date = 2004-07-15 pages = extension = .txt mime = text/plain words = 4010 sentences = 192 flesch = 55 summary = We examined serum samples obtained from 46 patients with SARS, 40 patients with non-SARS pneumonia, and 38 healthy individuals, by use of Western blotting (WB), enzyme-linked immunoassay (ELISA), and immunofluorescence assay, using both native and bacterially produced antigens of the virus. Both the humoral and cellular arms of the adaptive immune response are presumed to be important in controlling 2 or preparation U reacted with a serum sample from a patient with SARS showing the highly reactive antigens-nucleocapsid (N) 1, N2, and N3-in the former. Second, we made a recombinant antigen of the N-terminal half of the N protein (rNa), and, when it was used in an IgG ELISA, we found results almost identical to those found with the crude viral extract (89% sensitivity and 94%-95% specificity), including 4 negative cases in common ( figure 2) . cache = ./cache/cord-277735-a9gkath5.txt txt = ./txt/cord-277735-a9gkath5.txt === reduce.pl bib === id = cord-006828-i88on326 author = nan title = Abstracts DGRh-Kongress 2013 date = 2013-09-15 pages = extension = .txt mime = text/plain words = 30772 sentences = 2576 flesch = 52 summary = Comparing gene expression profiles of yellow fever immunized individuals and active SLE patients it was possible to identify a "common" and an "autoimmune-specific" IFN signature. The inflammatory and profibrotic effects upon Aab stimulation in vitro, and their associations with clinical findings suggest a role for autoantibody-mediated activation of immune cells mediated through the AT1R and ETAR in the pathogenesis or even the onset of the disease. This study was aimed to investigate the humoral and cellular immune response to VZV including assessment of IgG-anti-VZV avidity and VZV-specific reactivity of lymphocytes in RA (n=56) or JIA patients (n=75) on different treatments, including biologic agents, such as anti-tumor-necrosis-factor(TNF)-alpha or anti-interleukin-6 (IL-6) receptor inhibition (tocilizumab), compared to 37 healthy adults (HA) and 41 children (HC). Production of cytokines by B cells in response to TLR9 stimulation inversely correlates with disease activity in SLE-patients cache = ./cache/cord-006828-i88on326.txt txt = ./txt/cord-006828-i88on326.txt === reduce.pl bib === === reduce.pl bib === id = cord-262762-mgegswvn author = Callebaut, P. title = Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea date = 1982-09-30 pages = extension = .txt mime = text/plain words = 3605 sentences = 179 flesch = 50 summary = Abstract An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the coronavirus-like agent in feces of pigs naturally affected with porcine epidemic diarrhea (PED) or experimentally infected with the CV777 isolate. Control fecal samples, used to determine the limit between positive and negative absorbance values in ELISA, consisted of 25 non-diarrheal specimens obtained from conventional pigs of all ages. A total of 62 serum samples, to be examined for antibody by ELISA blocking, were collected from four CDCD piglets and seven conventional experimental fattening pigs prior to inoculation with CV777 and at various intervals between 7 and 81 days thereafter. To compare the sensitivity of EM and ELISA for the detection of CVLA, the specimens of fecal material and intestinal contents obtained from experimentally infected CDCD piglets and experimental fattening pigs as described in the section "Specimens from experimental pigs", were examined for coronavirus by EM. cache = ./cache/cord-262762-mgegswvn.txt txt = ./txt/cord-262762-mgegswvn.txt === reduce.pl bib === id = cord-022310-yc6xtw0s author = Lappin, Michael R. title = Microbiology and Infectious Disease date = 2011-12-15 pages = extension = .txt mime = text/plain words = 14109 sentences = 913 flesch = 39 summary = 47 Because of these findings, it is currently recommended to combine serologic test results with those of blood culture or PCR assay results when evaluating clinically ill cats for bartonellosis. Because serologic test results do not accurately correlate with presence of bacteremia and individual culture or PCR assay results can be falsely negative, there is no indication for testing healthy cats for Bartonella spp. T. gondii-specific IgM is detectable in serum by ELISA in approximately 80% of subclinically ill cats 2 to 4 weeks after experimental induction of toxoplasmosis; these titers generally are negative less than 16 weeks after infection. The author, however, has demonstrated IgG antibody titers greater than 1 : 16,384 in subclinically ill cats 5 years after experimental induction In chronic disease or asymptomatic carriers, demonstration of organisms is unreliable, and a tentative diagnosis is based on clinical signs and a positive titer. gondii-specific antibodies can also be detected in the serum, CSF, and aqueous humor of healthy, infected animals, one cannot assay results in clinically ill dogs, E. cache = ./cache/cord-022310-yc6xtw0s.txt txt = ./txt/cord-022310-yc6xtw0s.txt === reduce.pl bib === id = cord-022353-q2k2krnm author = W. Quimby, Fred title = Clinical Chemistry of the Laboratory Mouse date = 2007-09-02 pages = extension = .txt mime = text/plain words = 30195 sentences = 1702 flesch = 48 summary = Assessment of long-term average blood glucose levels in mice is also available by RIAs measuring glycosylated hemoglobin and glycosylated serum proteins (collectively known as fructosamines) (Gould et al. Leptin resistance, a common feature of obesity in mice and humans, has also been shown to result, in part, from the shedding of membrane-bound hepatic leptin receptors into the plasma, where soluble receptors modulate circulating leptin levels and possibly its biologic activity (Cohen et al. d. OTHER ANALYTES ASSOCIATED WITH LIPID METABOLISM AND ATHEROSCLEROSIS IN MICE ELISA kits are commercially available for the quantitation of many mouse coagulation proteins including: fibrinogen, factor VII, d-dimer, tissue factor, and von Willebrand's factor antigen. The ability of the first component of complement, C1, to bind specific sites on the heavy chain of mouse IgG2b and activate a sequence of reactions leading to production of a molecular unit capable of lysing a target cell membrane has established the complement system as the primary mediator of antibody-antigen reactions. cache = ./cache/cord-022353-q2k2krnm.txt txt = ./txt/cord-022353-q2k2krnm.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-276989-441aclcc author = Liu, Jianbo title = Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum date = 2020-03-12 pages = extension = .txt mime = text/plain words = 4023 sentences = 213 flesch = 54 summary = title: Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum On the strip, a gold-labeled anti-SIgA SC mAb was applied to a conjugate pad; purified PEDV particles and goat anti-mouse antibodies were blotted onto a nitrocellulose membrane to form the test and control lines, respectively. Therefore, the aim of this study was to develop a simple and rapid immunochromatographic test strip incorporating a colloidal gold-labeled anti-SIgA SC mAb probe for the detection of anti-PEDV-specific SIgA in swine, and to compare its performance with an indirect SIgA ELISA based on the whole PEDV virus (Cong et al., 2019) . To test for colostrum, we diluted the samples 1: 20 in sample buffer and mixed thoroughly then, 100 μL of the solution was dispensed onto the sample pad well of the strip apparatus to determine the presence of PEDV specific SIgA, as described above. cache = ./cache/cord-276989-441aclcc.txt txt = ./txt/cord-276989-441aclcc.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-275793-k0uvqcmp author = Xia, Hongyan title = A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies date = 2010-04-18 pages = extension = .txt mime = text/plain words = 2648 sentences = 154 flesch = 53 summary = title: A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies This study describes a novel blocking microsphere-based immunoassay for highly sensitive and specific detection of antibodies against bovine viral diarrhoea virus (BVDV). The objectives of this study were to develop a blocking microsphere-based immunoassay (bMIA) for detection of antibodies against BVDV, and to compare the performance of the assay with a commercial ELISA kit. This study describes the development and evaluation of a microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies. The diagnostic performance of the new bMIA was compared to that of a commercial blocking ELISA system, by testing a large panel of 509 bovine sera. A simple, rapid and reliable enzyme-linked immunosorbent assay for the detection of bovine virus diarrhoea virus (BVDV) specific antibodies in cattle serum, plasma and bulk milk cache = ./cache/cord-275793-k0uvqcmp.txt txt = ./txt/cord-275793-k0uvqcmp.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-281393-96j70n2z author = Capai, L. title = Seroprevalence of SARS-CoV-2 IgG antibodies, in Corsica (France), April and June 2020. date = 2020-09-30 pages = extension = .txt mime = text/plain words = 3684 sentences = 262 flesch = 60 summary = A minimum sample size of 1814 was calculated assuming an a priori 5% IgG anti-SARS-CoV-2 seroprevalence (Salje et al., 2020) , a confidence in the estimate of 95%, a maximum allowable error in the prevalence of 1%, and a Corsican population size of 344,679 habitants based on the latest French census data (INSEE, 2020). Residual sera obtained from persons of all ages were tested for the presence of anti-SARS-CoV-2 IgG using the EUROIMMUN enzyme immunoassay kit for semiquantitative detection of IgG antibodies against S1 domain of viral spike protein (ELISA-S) (reference: In all samples with a ratio ≥ 0.8, neutralizing antibodies were detected using a VNT as previously described (Gallian et al., 2020) . To the best of our knowledge this is the first study describing the prevalence of SARS-CoV-2 antibodies in a representative sample of Corsican patients having carried out a blood analysis in biological laboratories after the COVID19 epidemic period. cache = ./cache/cord-281393-96j70n2z.txt txt = ./txt/cord-281393-96j70n2z.txt === reduce.pl bib === id = cord-262268-gm99cadh author = Wang, Jingqiang title = Assessment of Immunoreactive Synthetic Peptides from the Structural Proteins of Severe Acute Respiratory Syndrome Coronavirus date = 2003-12-01 pages = extension = .txt mime = text/plain words = 4027 sentences = 189 flesch = 49 summary = Consequently, we thoroughly investigated the immunoreactivities with patient sera of a series of synthesized peptides from SARS-coronavirus structural proteins. Results: Four epitopic sites, S599, M137, N66, and N371-404, located in the SARS-coronavirus S, M, and N proteins, respectively, were detected by screening synthesized peptides. The peptides representing the COOH terminus of the N protein, in particular N371 and N385, had high absorbance/cutoff value ratios with the highest positive detection rate and the lowest hydrophobicity score among all of the synthesized peptides (Fig. 1C, and Fig. 3 in the online Data Supplement). The other 17 peptides reacted only slightly with the sera from SARS patients and gave low detection rates, suggesting that the regions of the S protein covered by these peptides have no epitopic site. The patient sera preincubated with 4 mg/L S599 or N385 gave a 25-30% lower response in the ELISA (data not shown), suggesting that the two peptides could compete with SARS coronavirus for binding to the antibodies in SARS serum. cache = ./cache/cord-262268-gm99cadh.txt txt = ./txt/cord-262268-gm99cadh.txt === reduce.pl bib === id = cord-254384-mwzz1db5 author = Lu, Guilan title = Large-scale seroprevalence analysis of human metapneumovirus and human respiratory syncytial virus infections in Beijing, China date = 2011-02-10 pages = extension = .txt mime = text/plain words = 3978 sentences = 242 flesch = 57 summary = To assess the seroprevalence of hMPV infection in China, we tested a total of 1,156 serum specimens for the presence of anti-hMPV IgG antibody in children and adults free of acute respiratory illness in Beijing, China by using hMPV nucleocapsid (N) protein as an antigen. To assess the seroprevalence of hMPV infection in China, we used hMPV N protein as an antigen to test serum samples for the presence of anti-hMPV IgG antibody in children and adults free of acute respiratory illness in Beijing, China. Lower seropositive rates and geometric mean titer (GMT) of anti-hMPV IgG were observed in children aged six months to six years when compared to hRSV. To test the specificity of the ELISA methods established in this study, the reactions of mouse sera against influenza virus A (subtypes H1-H16), human coronaviruses (229E, HKU1 and NL63), and polyomavirus JC against hMPV and hRSV N protein were evaluated. cache = ./cache/cord-254384-mwzz1db5.txt txt = ./txt/cord-254384-mwzz1db5.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-251943-jzaeaxam author = Zhang, Jian‐San title = A serological survey on neutralizing antibody titer of SARS convalescent sera date = 2005-08-24 pages = extension = .txt mime = text/plain words = 2542 sentences = 139 flesch = 50 summary = A seroepidemiologic study was conducted in North China in 2003 to determine the neutralizing antibody titer of severe acute respiratory syndrome (SARS) convalescent sera. A total of 99 SARS convalescent serum samples were collected from patients from the Inner Mongolia Autonomous Region, Hebei Province, and Beijing 35–180 days after the onset of symptoms. To gain a comprehensive understanding of the antibody to SARS-CoV, we report the anti-SARS antibody titer of 87 SARS convalescent sera determined by neutralization assay. These 87 serum samples were confirmed to be positive for anti-SARS antibodies with the combination of ELISA, neutralization, and Western blot, so they were pooled to form a convalescent sera database for the further analysis of neutralizing antibody titer. The anti-SARS neutralizing antibody titer of 87 positive convalescent sera was analyzed quantitatively by the neutralization assay. In our laboratory, a combination of ELISA, neutralization assay, and Western blot were performed on 99 SARS convalescent sera. cache = ./cache/cord-251943-jzaeaxam.txt txt = ./txt/cord-251943-jzaeaxam.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-279406-wwdqh9qs author = Guzman, Norberto A. title = A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy date = 2020-07-30 pages = extension = .txt mime = text/plain words = 17172 sentences = 835 flesch = 33 summary = To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses. It would be beneficial to have a sample processing method before separation, to isolate and concentrate the intended viruses or EVs. Immunoaffinity capillary electrophoresis has already been proven to be a useful technology to isolate, separate, and quantify cell-free molecules of biological interest based on the specificity and selectivity not only of antibody reagents, but also of lectin and aptamer reagents, quantifying molecules ranging from microgram/milliliter to femtogram/milliliter [25, 54, 55, 57, 75] . cache = ./cache/cord-279406-wwdqh9qs.txt txt = ./txt/cord-279406-wwdqh9qs.txt === reduce.pl bib === id = cord-254318-w8wrn9lx author = Díez, José-María title = Currently available intravenous immunoglobulin contains antibodies reacting against severe acute respiratory syndrome coronavirus 2 antigens date = 2020-05-13 pages = extension = .txt mime = text/plain words = 2687 sentences = 167 flesch = 48 summary = MATERIAL & METHODS: Gamunex(®)-C and Flebogamma(®) DIF (Grifols) intravenous immunoglobulin (IVIG) products were tested using ELISA techniques for antibodies against several antigens of human common betacoronaviruses that may crossreact with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. Gamunex R -C (Grifols Therapeutics, Inc., NC, USA) and Flebogamma R DIF (Instituto Grifols S.A., Barcelona, Spain) IVIGs were tested for crossreactivity against several betacoronaviruses, including SARS-CoV, MERS-CoV and SARS-CoV-2 antigens, using ELISA techniques. Even with this uncertainty, in the context of the current health emergency (pandemic), the potential of IVIG as a therapy for COVID-19 is already being evaluated in a number of studies involving patients with severe SARS-CoV-2 viral infections including pneumonia [28] [29] [30] . • This is the first time that currently available intravenous immunoglobulins have been reported to contain antibodies that crossreact against antigens of SARS-CoV-2 and other coronaviruses. cache = ./cache/cord-254318-w8wrn9lx.txt txt = ./txt/cord-254318-w8wrn9lx.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-281760-34wuttqw author = Pereira, E.P.V. title = Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review date = 2019-05-22 pages = extension = .txt mime = text/plain words = 9686 sentences = 431 flesch = 42 summary = Considering the fast development of IgY technology, this work aims to review its applications in human and animal health, in addition to increasing the potential use of these antibodies among researchers and consequently promoting the reduced use of non-invasive procedures on animals. extracted IgY from hens immunized with the recombinant protein FanC, from enterotoxigenic Escherichia coli (ETEC) and these antibodies bound specifically to FanC in ELISA, Western blot and Dot-blotting [59] , demanding, thus, more investigations to evaluate its viability as a potential immunotherapeutic compound. Anti-DENV2 IgY produced in goose was able to neutralize the virus in vitro and in vivo without binding to Fcγ receptors on myeloid cells and generating ADE (antibody dependent enhancement) in mice [57] . Hen egg yolk antibodies (IgY), production and use for passive immunization against bacterial enteric infections in chicken: a review Preventive effect of anti-VacA egg yolk immunoglobulin (IgY) on Helicobacter pylori-infected mice cache = ./cache/cord-281760-34wuttqw.txt txt = ./txt/cord-281760-34wuttqw.txt === reduce.pl bib === === reduce.pl bib === id = cord-301313-9595vm0k author = OKBA, NISREEN M.A. title = SARS-CoV-2 specific antibody responses in COVID-19 patients date = 2020-03-20 pages = extension = .txt mime = text/plain words = 4271 sentences = 248 flesch = 55 summary = Here, we describe development of serological assays for the detection of virus neutralizing antibodies and antibodies to the nucleocapsid (N) protein and various spike (S) domains including the S1 subunit, and receptor binding domain (RBD) of SARS-CoV-2 in ELISA format. Using a wellcharacterized cohort of serum samples from PCR-confirmed SARS-CoV-2 and patients PCR-confirmed to be infected with seasonal coronaviruses and other respiratory pathogens, we validated and tested various antigens in different platforms developed in-house as well as a commercial platform. We evaluated SARS-CoV-2 specific antibody responses in severe and mild cases using serum samples collected at different times post-disease onset from three French PCR-confirmed CoVID-19 patients. We tested sera for SARS-CoV-2 specific antibodies using different ELISAs. Following infections, all three patients seroconverted between days 13 and 21 post onset of disease (Figure 1) , and antibodies were elicited against the SARS-CoV-2 S and S1 subunit including the N-terminal (S1 A ) domain and the receptor binding domain (RBD). cache = ./cache/cord-301313-9595vm0k.txt txt = ./txt/cord-301313-9595vm0k.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-001521-l36f1gp7 author = nan title = Oral and Poster Manuscripts date = 2011-04-08 pages = extension = .txt mime = text/plain words = 183363 sentences = 11362 flesch = 53 summary = The IC 50 values determined in functional NI assays provide valuable information for detection of resistant viruses, but should not be used to draw direct correlations with drug concentrations needed to inhibit virus replication in the infected human host, as clinical data to support such inferences are inadequate. • Standardized reagents and protocols • Choice of detection technology • Simple instrumentation requirements • High sensitivity for use with low virus concentrations • Compatibility with batch-mode processing and largescale assay throughput • Broad specificity of influenza detection • Flexibility in assay format • Additional NA assay applications -cell-based viral assays, screening for new NIs, detection of NA from other organisms Functional neuraminidase inhibition assays enable detection of any resistance mutation and are extremely important in conjunction with sequence-based screening assays for global monitoring of virus isolates for NI resistance mutations, including known and new mutations. Such new assays need to include methods to measure local antibodies and virus-specific lymphocytes, especially in the case of live attenuated influenza vaccines, because of their potential to induce such broad-based immune responses. cache = ./cache/cord-001521-l36f1gp7.txt txt = ./txt/cord-001521-l36f1gp7.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-015147-h0o0yqv8 author = nan title = Oral Communications and Posters date = 2014-09-12 pages = extension = .txt mime = text/plain words = 73711 sentences = 3862 flesch = 43 summary = Cyclooxygenases (COX) catalyze the first step in the synthesis of prostaglandins (PG) from arachidonic acid.COX-1 is constitutively expressed.The COX-2 gene is an immediate early-response gene that is induced by variety of mitogenic and inflammatory stimuli.Levels of COX-2 are increased in both inflamed and malignant tissues.In inflamed tissues, there is both pharmacological and genetic evidence that targeting COX-2 can either improve (e.g., osteoarthritis) or exacerbate symptoms (e.g., inflammatory bowel disease).Multiple lines of evidence suggest that COX-2 plays a significant role in carcinogenesis.The most specific data that support a cause-and effect relationship between COX-2 and tumorigenesis come from genetic studies.Overexpression of COX-2 has been observed to drive tumor formation whereas COX-2 deficiency protects against several tumor types.Selective COX-2 inhibitors protect against the formation and growth of experimental tumors.Moreover, selective COX-2 inhibitors are active in preventing colorectal adenomas in humans.Increased amounts of COX-2-derived PGE2 are found in both inflamed and neoplastic tissues.The fact that PGE2 can stimulate cell proliferation, inhibit apoptosis and induce angiogenesis fits with evidence that induction of COX-2 contributes to both wound healing and tumor growth.Taken together, it seems likely that COX-2 induction contributes to wound healing in response to injury but reduces the threshold for carcinogenesis. cache = ./cache/cord-015147-h0o0yqv8.txt txt = ./txt/cord-015147-h0o0yqv8.txt === reduce.pl bib === id = cord-311349-145kwny3 author = Mariani, Stefano title = Surface plasmon resonance applications in clinical analysis date = 2014-02-25 pages = extension = .txt mime = text/plain words = 13425 sentences = 630 flesch = 39 summary = In the last 20 years, surface plasmon resonance (SPR) and its advancement with imaging (SPRi) emerged as a suitable and reliable platform in clinical analysis for label-free, sensitive, and real-time monitoring of biomolecular interactions. The advantages brought about by current SPR technology include real-time monitoring of the analyte/molecular markers, label free and parallel analysis (with SPRi), minimal sample pretreatment, quantitative response, and very good sensitivity and reproducibility, (reported detection limits are in atto-or femtomolar ranges and coefficient of variations below 10 %). Preventing nonspecific adsorption of biomolecules (e.g., protein) on the SPR sensing surface is another key-step for the development of specific biosensor, with real application to clinical diagnostics where complex matrices (such as serum, blood, and urine) are analyzed. cache = ./cache/cord-311349-145kwny3.txt txt = ./txt/cord-311349-145kwny3.txt === reduce.pl bib === id = cord-313517-5ipj2z86 author = Fung, Joshua title = Antigen Capture Enzyme-Linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans date = 2019-09-14 pages = extension = .txt mime = text/plain words = 2710 sentences = 149 flesch = 51 summary = Though the gold standard for diagnosing MERS-CoV infection in humans is still nucleic acid amplification test (NAAT) of the up-E region, an antigen capture enzyme-linked immunosorbent assay (ELISA) could also be of use for early diagnosis in less developed locations. In the present method, a step-by-step guide to perform a MERS-CoV nucleocapsid protein (NP) capture ELISA using two NP-specific monoclonal antibodies is provided for readers to develop their in-house workflow or diagnostic kit for clinical use and for mass-screening project of animals (e.g., dromedaries and bats) to better understand the spread and evolution of the virus. Nucleic acid amplification test (NAAT, e.g., real-time reverse transcription quantitative polymerase chain reaction [real-time RT-qPCR]), virus isolation, transmission electron microscopy, immunohistochemistry, and serological methods (e.g., antigen capture enzyme-linked immunosorbent assay [ELISA] and immunofluorescence assay [IFA] ) have been developed and used for MERS-CoV diagnosis [2] [3] [4] [5] [6] [7] . cache = ./cache/cord-313517-5ipj2z86.txt txt = ./txt/cord-313517-5ipj2z86.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-301175-6alsigxk author = Okda, Faten title = Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus date = 2015-08-01 pages = extension = .txt mime = text/plain words = 8275 sentences = 411 flesch = 46 summary = title: Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus Therefore, the objective of this study was to develop and validate multiple improved serological assays for PEDV, including an indirect ELISA (iELISA); a highly specific monoclonal antibody-based blocking ELISA (bELISA); fluorescent microsphere immunoassays (FMIA) that can be multiplexed to monitor exposure to multiple antigens and pathogens simultaneously; and a fluorescent focus neutralization assay (FFN) to measure functional virus neutralizing antibodies. RESULTS: A recombinant North American nucleoprotein (NP) based iELISA was developed and validated along with a bELISA using newly developed PEDV-NP specific biotinylated monoclonal antibodies (mAbs) and an FMIA using magnetic beads coupled with expressed NA PEDV-NP. In this study, we report the adaptation of a recombinant, highly purified, NA PEDV-NP antigen to the development of iELISA, bELISA and FMIA platforms for the detection of PEDV antibodies in serum. cache = ./cache/cord-301175-6alsigxk.txt txt = ./txt/cord-301175-6alsigxk.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-006860-a3b8hyyr author = nan title = 40th Annual Meeting of the GTH (Gesellschaft für Thrombose- und Hämostaseforschung) date = 1996 pages = extension = .txt mime = text/plain words = 90660 sentences = 5152 flesch = 50 summary = Dept of Pediatrics, University Hospitals Kiel and Mtinster, Germany Resistance to activated protein C (APCR), in the majority of cases associated with the Arg 506 Gin point mutation in the factor V gene is present in more than 50 % of patients < 60 years of age with unexplained thrombophilia. The regular APC resistance test is not applicable to plasma from Orally anticoagulated (OAC) or heparinized patients due to decreased levels of vitamin K-dependent clotting factors and to thrombin inhibition by antithrombin, respectively. On admission an extensive coagulation screen yielded the following results (n/normal, t/elevated, I/reduced, +/positive, -/negative): PT t, aPTT t, Tr n, factor II, V, VIII n, factor VII, IX, XI, XII /,, fibrinogan t, ATIII n, protein C, S *, activated protein C sensitivity ratio 1.92 ($), FV-Leidenmutation PCR -, fibrinolytic system n, TAT t, Ft÷2 t, lupus anticoagulant +, heparin induced platelet antibodies +; no diagnosis of a specific autoimmuna disorder could be made. cache = ./cache/cord-006860-a3b8hyyr.txt txt = ./txt/cord-006860-a3b8hyyr.txt === reduce.pl bib === === reduce.pl bib === === reduce.pl bib === id = cord-317123-0tdfvlqd author = Tan, Xiaotian title = Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples date = 2020-04-21 pages = extension = .txt mime = text/plain words = 4154 sentences = 220 flesch = 52 summary = Here, we present a microfluidic ELISA technology for rapid (15-20 minutes), quantitative, sensitive detection of SARS-CoV-2 biomarkers using SARS-CoV-2 specific IgG and viral antigen – S protein in serum. We also characterized various humanized monoclonal IgG, and identified a candidate with a high binding affinity towards SARS-CoV-2 S1 protein that can serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. In this work, we present a microfluidic ELISA technology for rapid (15-20 minutes), quantitative, and sensitive detection of SARS-CoV-2 biomarkers using SARS-CoV-2 specific IgG and viral antigen -S protein, both of which are spiked in serum, as a model system. For the anti-S1 IgG detection experiments (see Figure S2 (B) for the detailed protocol), various concentrations of monoclonal antibodies were prepared by diluting the stock solutions with 50 times diluted human serum (the serum was diluted with 1× reagent diluent, which correlates to 1% BSA). cache = ./cache/cord-317123-0tdfvlqd.txt txt = ./txt/cord-317123-0tdfvlqd.txt === reduce.pl bib === id = cord-312456-6lxc2rj2 author = Soltan, Mohamed A. title = Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species date = 2016-05-11 pages = extension = .txt mime = text/plain words = 4209 sentences = 218 flesch = 53 summary = title: Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species The aim of this investigation was to evaluate a commercially available RT-iiPCR assay for RVA detection in feces from different animal species. Therefore, the aim of our investigation was to evaluate a recently available insulated isothermal RT-PCR (RT-iiPCR) reagent set (POCKIT TM Rotavirus A Reagent Set, GeneReach USA, Lexington, MA, USA) with use of a portable PCR machine, which could potentially be used for point-of-need detection for RVA in the feces of different animal species. Additionally, the sensitivity of the rotavirus RT-iiPCR reagent set was evaluated by comparison with the commercially available rtRT-PCR assay using 10-fold serial dilutions of nucleic acid extracted from a positive bovine clinical sample. There was a significant difference in the number of positive samples detected with the in-house rtRT-PCR assay versus the other two molecular tests. cache = ./cache/cord-312456-6lxc2rj2.txt txt = ./txt/cord-312456-6lxc2rj2.txt === reduce.pl bib === id = cord-022653-qa1uph35 author = nan title = Poster Discussion Session PDS date = 2017-08-30 pages = extension = .txt mime = text/plain words = 58292 sentences = 3300 flesch = 53 summary = 0206 | G protein coupled receptor kinase 2 (GRK2) regulates endothelial permeability induced by Bradykinin 0208 | Pharmacokinetics (PK) and pharmacodynamics (PD) of c1 esterase inhibitor of chronic urticaria challenges most commonly identified were the following: time of onset of disease; frequency/duration of and provoking factors for wheals; diurnal variation; occurrence in relation to weekends, holidays, and foreign travel; shape, size, and distribution of wheals; associated angioedema; associated subjective symptoms of lesions; family and personal history regarding urticaria, atopy; previous or current allergies, infections, internal diseases, or other possible causes; psychosomatic and psychiatric diseases; surgical implantations and events during surgery; gastric/ intestinal problems; induction by physical agents or exercise; use of drugs; food allergies; relationship to the menstrual cycle; smoking habits; type of work, hobbies; stress; quality of life and emotional impact; previous therapy and response to therapy, and previous diagnostic procedures/results. cache = ./cache/cord-022653-qa1uph35.txt txt = ./txt/cord-022653-qa1uph35.txt === reduce.pl bib === id = cord-318444-sgm24q1i author = Walter, Justin D. title = Sybodies targeting the SARS-CoV-2 receptor-binding domain date = 2020-05-16 pages = extension = .txt mime = text/plain words = 5902 sentences = 416 flesch = 49 summary = Two independently prepared RBD constructs were used for in vitro sybody selections, and resulting single clones that could bind the full spike ectodomain were sequenced, expressed, and purified. Six unique sybodies show favorable binding affinity to the SARS-CoV-2 spike, and five of these were also found to substantially attenuate the interaction between the viral RBD and human ACE2. While this purified pre-fusion spike (PFS) had not yet been available for binder selections and characterization by grating-coupled interferometry, it was used to conduct ELISAs in order to identify selected sybodies which recognize the RBD in the pre-fusion context (see below). Since virulence of SARS-CoV-2 is dependent on the ability of the viral RBD to bind to human ACE2 (hACE2), we sought to determine which of the 57 selected sybodies that were well-behaved upon purification could inhibit interaction between the isolated RBD and purified hACE2. cache = ./cache/cord-318444-sgm24q1i.txt txt = ./txt/cord-318444-sgm24q1i.txt === reduce.pl bib === id = cord-026559-xx52u01h author = Tripathi, Siddhartha title = Blood Plasma Microfluidic Device: Aiming for the Detection of COVID-19 Antibodies Using an On-Chip ELISA Platform date = 2020-06-10 pages = extension = .txt mime = text/plain words = 2010 sentences = 125 flesch = 52 summary = title: Blood Plasma Microfluidic Device: Aiming for the Detection of COVID-19 Antibodies Using an On-Chip ELISA Platform We propose to first separate plasma from whole human blood using a microfluidic device and subsequently perform the detection of antibodies in the separated plasma using a semi-automated on-chip ELISA. The reported plasma separation microdevice is not only an alternate to the centrifuge, but it can also be easily integrated with a biosensing platform/detection technology (for example, ELISA) and result in a point-of-care device. (2020) have reported successful detection of SARS-CoV-2 virus with high sensitivity in swab specimens Fig. 1 a Blood plasma microdevice design and zoomed view at the junction. Herein, we propose the integration of sandwich ELISA (enzyme-linked immunoabsorbent assay) with the blood plasma separation microdevice to detect COVID-19 antibodies after minor modifications in the design. b Experi-mental sandwich ELISA: showing steps to identify the presence of SARS-CoV-2 (COVID-19) antibodies present in blood plasma separated and flows towards the plasma outlet reservoir. cache = ./cache/cord-026559-xx52u01h.txt txt = ./txt/cord-026559-xx52u01h.txt === reduce.pl bib === id = cord-313193-q5zeoqlb author = Carrat, F. title = Seroprevalence of SARS-CoV-2 among adults in three regions of France following the lockdown and associated risk factors: a multicohort study. date = 2020-09-18 pages = extension = .txt mime = text/plain words = 4076 sentences = 265 flesch = 57 summary = Methods Participants in a survey on COVID-19 from an existing consortium of three general adult population cohorts living in the Ile-de-France (IDF) or Grand Est (GE), two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA), with a low rate, were asked to take a dried-blood spot (DBS) for anti-SARS-CoV-2 antibodies assessment. Participants in a survey on COVID-19 from an existing consortium of three general adult population cohorts living in the Ile-de-France (IDF) or Grand Est (GE) -two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA) -with a low rate, were asked to take a dried-blood spot (DBS) for anti-SARS-CoV-2 antibodies assessment. Participants with a positive ELISA-S had a higher rate of self-reported symptoms than those with negative tests except for skin lesions (table 2) is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint cache = ./cache/cord-313193-q5zeoqlb.txt txt = ./txt/cord-313193-q5zeoqlb.txt === reduce.pl bib === id = cord-317057-c2bwky6e author = Pickering, S. title = Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date = 2020-06-04 pages = extension = .txt mime = text/plain words = 5304 sentences = 269 flesch = 53 summary = A highly specific in-house ELISA was developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs) on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. Accordingly, we developed a highly specific semi-quantitative 4 ELISA for the detection of anti-spike (S), -S receptor binding domain (RBD) and -N antibodies, and used this to cross-evaluate ten commercial antibody tests (seven lateral flow immunoassays (LFIAs), one chemiluminescent assay and two ELISAs) on a collection of 110 serum samples from confirmed RNA positive patients, and 50 pre-pandemic samples from March 2019. With no existing gold standard for the assessment of the serological response to SARS-CoV-2, we started by comparing commercial serological assays with the best performing configuration of the in-house ELISA (detection of anti-S IgM and IgG antibodies), which was also most likely to represent antibodies detected by the commercial tests. cache = ./cache/cord-317057-c2bwky6e.txt txt = ./txt/cord-317057-c2bwky6e.txt === reduce.pl bib === id = cord-327890-ocisq7e4 author = Pallett, Scott J C title = Point-of-care serological assays for delayed SARS-CoV-2 case identification among health-care workers in the UK: a prospective multicentre cohort study date = 2020-07-24 pages = extension = .txt mime = text/plain words = 5947 sentences = 271 flesch = 42 summary = In phase 1, two point-of-care lateral flow serological assays, the Onsite CTK Biotech COVID-19 split IgG/IgM Rapid Test (CTK Bitotech, Poway, CA, USA) and the Encode SARS-CoV-2 split IgM/IgG One Step Rapid Test Device (Zhuhai Encode Medical Engineering, Zhuhai, China), were evaluated for performance against a laboratory immunoassay (EDI Novel Coronavirus COVID-19 IgG ELISA kit [Epitope Diagnostics, San Diego, CA, USA]) in 300 samples from health-care workers and 100 pre-COVID-19 negative control samples. These include point-of-care molecular platforms for acute phase testing, and laboratory ELISA or lateral flow serological assays for antibodies specific to SARS-CoV-2 for delayed case identification. To derive a measure of sensitivity, the results of lateral flow serological assays and ELISA were compared in 300 health-care workers who had previously received PCR testing (AusDiagnostics, Sydney, Australia) at initial presentation with COVID-19 symptoms. cache = ./cache/cord-327890-ocisq7e4.txt txt = ./txt/cord-327890-ocisq7e4.txt === reduce.pl bib === id = cord-334165-7gfk554m author = Stadlbauer, Daniel title = SARS‐CoV‐2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup date = 2020-04-17 pages = extension = .txt mime = text/plain words = 5286 sentences = 414 flesch = 69 summary = Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two‐stage ELISA for high‐throughput screening of human serum samples for antibodies binding to the spike protein of SARS‐CoV‐2 We reported in our earlier work that individuals not exposed to SARS-CoV-2 are completely naïve to the spike protein, and their serum samples show little or no reactivity in an ELISA (Amanat et al., 2020) . We developed this as a two-stage ELISA in which the first stage ('a' steps below) includes relatively high-throughput screening of samples in a single serum dilution against the RBD (which expresses very well and therefore can be produced in greater quantities). This is followed by a second stage ('b' steps below) in which positive samples from the first stage undergo a confirmatory ELISA against the full-length spike protein (which is harder to express; therefore there is usually less available). i. Thaw the required number of vials of antigen (SARS-CoV-2 RBD protein) to coat 96-well microtiter ELISA plates at a concentration of 2 μg/ml. cache = ./cache/cord-334165-7gfk554m.txt txt = ./txt/cord-334165-7gfk554m.txt === reduce.pl bib === id = cord-320559-up1q3k6q author = Dortmans, J.C.F.M. title = Porcine epidemic diarrhea virus (PEDV) introduction into a naive Dutch pig population in 2014 date = 2018-05-24 pages = extension = .txt mime = text/plain words = 4216 sentences = 207 flesch = 57 summary = Porcine epidemic diarrhea virus (PEDV) is the highly contagious, causative agent of an economically important acute enteric disease in pigs of all ages. In total, 838 blood samples from sows from 267 farms and 101 samples from wild boars were collected from May till November 2014 and tested for antibodies against PEDV by ELISA. The number of required blood samples from animals and farms to estimate the seroprevalence of PEDV in Dutch sow herds was calculated based on the following assumptions: PEDV is highly contagious and no vaccination against this virus was carried out in the Netherlands. For the detection of PEDV antibodies in serum samples an in-house indirect ELISA based on the viral spike (S) protein S1-part of the G2b strain GDU (Non-S-INDEL, GenBank KU985230.1) was used, similar as the ELISA previously described (Gerber et al., 2014) . cache = ./cache/cord-320559-up1q3k6q.txt txt = ./txt/cord-320559-up1q3k6q.txt === reduce.pl bib === id = cord-317026-9zgc6xrb author = Zhao, Shan title = Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus date = 2019-11-30 pages = extension = .txt mime = text/plain words = 6223 sentences = 317 flesch = 55 summary = In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure. On the other hand, serological assays can be used to support the diagnosis of a clinical ECoV infection by showing seroconversion or a significant increase in antibody titer in paired serum samples. For the six horses that did not show a significant rise in VNT, five serum samples collected at the acute stage already had high antibody levels as shown by ELISA and neutralization assay (mean OD value > 2.6, mean VNT > 9, Table S1 ). For the six horses that did not show a significant rise in VNT, five serum samples collected at the acute stage already had high antibody levels as shown by ELISA and neutralization assay (mean OD value > 2.6, mean VNT > 9, Table S1 ). cache = ./cache/cord-317026-9zgc6xrb.txt txt = ./txt/cord-317026-9zgc6xrb.txt === reduce.pl bib === id = cord-333026-9f6ecg30 author = Kompanikova, J. title = Microbiologic Methods in the Diagnostics of Upper Respiratory Tract Pathogens date = 2017-03-03 pages = extension = .txt mime = text/plain words = 1784 sentences = 100 flesch = 49 summary = Blood samples were simultaneously examined by the enzyme-linked immunosorbent assay (ELISA) and by the FilmArray Respiratory Panel for eight different pathogens in a total of 15 tests performed in nasopharyngeal swabs. Nonetheless, since most repiratory tract infections are viral in origin and there is no treatment available, the diagnosis provided by the FilmArray Panel does not provide any additional clinical benefit and thus should be used only whenever necessary on the individual basis. This method allows for identification of 21 different respiratory pathogens from a nasopharyngeal swab, 18 of viral etiology and three of bacterial origin (Idaho Technology 2007). The laboratory costs to run one examination with different methods showed that the FilmArray multiplex PCR respiratory panel is more expensive than the ELISA, HIT, and the cultivation. Since most URIs are viral in origin and there is no treatment available, the diagnosis provided by the FilmArray panel is not always necessary and the method should be used on an individual basis when clinically justified. cache = ./cache/cord-333026-9f6ecg30.txt txt = ./txt/cord-333026-9f6ecg30.txt === reduce.pl bib === id = cord-326232-668f53qc author = Toftaker, Ingrid title = Evaluation of a multiplex immunoassay for bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk against two indirect ELISAs using latent class analysis date = 2018-06-01 pages = extension = .txt mime = text/plain words = 5627 sentences = 307 flesch = 56 summary = The objective of this study was to estimate sensitivity and specificity across different cut-off values for the MVD-Enferplex BCV/BRSV multiplex, by comparing them to a commercially available ELISA, the SVANOVIR(®) BCV-Ab and SVANOVIR(®) BRSV-Ab, respectively. The aim of this study was to estimate the test sensitivity and specificity of the newly developed MVD-Enferplex BCV/BRSV multiplex across different cut-off values, for detection of antibodies in BTM. Estimates of test parameters and true prevalence in the two subpopulations across different cut-off values for the BCV multiplex and the BCV ELISA are presented in Table 5 . The chosen cut-off will Table 4 Results from the sensitivity analysis (BRSV): Median estimates and 95% posterior credibility intervals (PCI) of the sensitivity (Se) and specificity (Sp) of bulk tank milk BRSV multiplex and BRSV ELISA at the manufacturers' recommended cut-off (alternative 2, Fig. 1 ), for the conditionally independent (CID) model and conditionally dependent (COC) models where the covariance is expressed as proportions of maximum possible value. cache = ./cache/cord-326232-668f53qc.txt txt = ./txt/cord-326232-668f53qc.txt === reduce.pl bib === id = cord-334277-g3go3u02 author = Kovac, Marc title = EDTA-Anticoagulated Whole Blood for SARS-CoV-2 Antibody Testing by Electrochemiluminescence Immunoassay (ECLIA) and Enzyme-Linked Immunosorbent Assay (ELISA) date = 2020-08-14 pages = extension = .txt mime = text/plain words = 4725 sentences = 237 flesch = 53 summary = While lateral flow test formats can be utilized with whole blood and low sample volumes, their diagnostic characteristics are inferior to immunoassays based on chemiluminescence immunoassay (CLIA) or enzyme-linked immunosorbent assay (ELISA) technology. We addressed the suitability of EDTA-anticoagulated whole blood as an alternative sample material for antibody testing against SARS-CoV-2 by electro-CLIA (ECLIA; Roche, Rotkreuz, Switzerland) and ELISA (IgG and IgA; Euroimmun, Germany). In receiver-operating characteristic curve analysis, all three assays displayed comparable diagnostic accuracy (area under the curve (AUC)) using corrected whole blood and serum (AUCs: 0.97 for ECLIA and IgG ELISA; 0.84 for IgA ELISA). It can thus be concluded that the anti-SARS-CoV-2 antibody results in whole blood corrected for hematocrit with weakly and moderately positive findings are comparable to those obtained from serum. cache = ./cache/cord-334277-g3go3u02.txt txt = ./txt/cord-334277-g3go3u02.txt === reduce.pl bib === id = cord-317223-juw4xt8q author = Pedersen, Niels C. title = The causes of false-positives encountered during the screening of old-world primates for antibodies to human and simian retroviruses by ELISA date = 1986-11-30 pages = extension = .txt mime = text/plain words = 3923 sentences = 190 flesch = 51 summary = False-positive ELISA antibody tests were particularly common among sera from mandrills, crab-eating macaques, lion-tailed macaques, African green monkeys, and DeBrazza's and moustached guenons. False positive ELISA antibody tests, while sporadically encountered among most of the 50 species of oldworld primates, were especially prevalent in Mandrillus sphinx (mandrills), Macaca fasicularis (crab-eating macaques) , Macaca sifensus (lion-tailed macaques), Cercopithecus aethiops (African green monkeys), Cercopithecus neglectus (De-Brazza's guenons), Cercopithecus cephus (moustached guenons) and Miopithecus talapoin (talapoins) . Specific antibodies to HTLV-III were found in 23 monkeys, 4/7 sooty mangabys, 11/15 talapoins, 2/11 False-positive ELISA antibody reactions to one or more viruses were found in a low proportion of individuals from many different species of old-world primates (Table 1) . Studies with African green monkey sera established the cause of false-positive ELISA antibody tests in this species; sera appeared to contain a specific antibody or antibodies that was directed against cell-associated protein(s) that were co-purified with the various virus preparations. cache = ./cache/cord-317223-juw4xt8q.txt txt = ./txt/cord-317223-juw4xt8q.txt === reduce.pl bib === id = cord-305516-s1lvhknm author = Watanabe, Shumpei title = Epizootology and experimental infection of Yokose virus in bats date = 2008-09-11 pages = extension = .txt mime = text/plain words = 4133 sentences = 232 flesch = 55 summary = To reveal whether bats serve as an amplifying host for Yokose virus (YOKV), we conducted a serological survey and experimentally infected fruit bats with YOKV isolated from microbats in Japan. To detect antibodies against YOKV, we developed an enzyme-linked immunosorbent assay (ELISA) using biotinylated anti-bat IgG rabbit sera. To detect antibodies against YOKV, we developed an ELISA using biotinylated anti-bat IgG rabbit sera. Using the conventional ELISA, a serological survey was performed on bat serum samples collected from the Philippines and Malaysia. To obtain sera positive for anti-YOKV antibodies, two bats were immunized with inactivated and purified virus. Furthermore, the serum samples collected from bats which were obtained from Japan Zoos were also screened by ELISA using the JEV antigen. We developed an ELISA system using biotin-labeled anti-bat-IgG rabbit serum to detect antibodies against YOKV in bat sera and conducted serological surveys using this system. cache = ./cache/cord-305516-s1lvhknm.txt txt = ./txt/cord-305516-s1lvhknm.txt === reduce.pl bib === id = cord-318266-i8x80knq author = Böse, Reinhard title = Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot date = 1994-05-31 pages = extension = .txt mime = text/plain words = 3622 sentences = 220 flesch = 61 summary = title: Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. Sera tested were as follows: (I) sera from 106 horses not infected with Bubesia spp., i.e. horses from Germany tested negative in preliminary studies by IFAT at a serum dilution of 4 l/40 and by CFT at a serum dilution of < l/5; the results for these sera were used to calculate the diagnostic specificity of the ELISA and Western blot; (2) 71 sera from 15 horses experimentally infected with B. equi; these results were used to calculate the diagnostic sensitivity of the ELISA, Western blot, IFAT and CFT; (3) 76 sera from 13 horses experimentally infected with 3. cache = ./cache/cord-318266-i8x80knq.txt txt = ./txt/cord-318266-i8x80knq.txt === reduce.pl bib === id = cord-343185-lbmbp9ca author = Hansen, C. B. title = SARS-CoV-2 antibody responses determine disease severity in COVID-19 infected individuals date = 2020-07-29 pages = extension = .txt mime = text/plain words = 5349 sentences = 332 flesch = 51 summary = Here we have developed novel flexible ELISA-based assays for specific detection of SARS-CoV-2 antibodies against the receptor-binding domain (RBD): An antigen sandwich-ELISA relevant for large population screening and three isotype-specific assays for in-depth diagnostics. Detection of IgM, IgA and IgG antibodies against SARS-CoV-2 protein N was evaluated by analyzing 136 positive samples and 174 negative controls and ROC curve analyses were assessed to estimate the assay performance . To provide a better insight into antibody seroconversion during SARS-CoV-2 infection and reactivity against different locations on protein S and protein N, we conducted IgM, IgA and IgG detection in 90 positive samples against 14 protein fragments and short peptides located on the protein S and protein N structures, full-length RBD, protein S and protein N (Figure 2A ). We have developed an ELISA-based platform for detection SARS-CoV-2 antibodies comprising an indirect RBD S-ELISA for pan Ig detection and direct ELISAs for in-depth analyses of the IgM, IgA and IgG isotype responses towards RBD and protein N. cache = ./cache/cord-343185-lbmbp9ca.txt txt = ./txt/cord-343185-lbmbp9ca.txt === reduce.pl bib === id = cord-334968-gonx5taq author = Pignatelli, Jaime title = Lineage specific antigenic differences in porcine torovirus hemagglutinin-esterase (PToV-HE) protein date = 2013-12-23 pages = extension = .txt mime = text/plain words = 6772 sentences = 291 flesch = 50 summary = In the first case, the two PToV-HE lineages were detected even within the same animal at two sequential sampling time points, indicating that both PToV strains carrying different HE proteins coexisted on the same farm infecting the same piglets, and suggesting that the immune response generated against one PToV strain did not protect the animals against the infection by the other strain. Hence, in order to examine the potential existence of antigenic differences between the two lineages of PToV-HE proteins, piglet serum samples were analyzed with an HI assay to detect the presence of specific antibodies against the receptor binding domain that prevents the RBC hemagglutination. In addition, to study the antibody response against the whole protein by a different approach, soluble c-myc tagged HE52.7 and HE52.11 proteins were generated by rVV methodology to obtain highly purified coating antigens that were used in ELISA to test the same field serum samples. cache = ./cache/cord-334968-gonx5taq.txt txt = ./txt/cord-334968-gonx5taq.txt === reduce.pl bib === id = cord-321691-46la29tm author = Hsueh, Po-Ren title = SARS Antibody Test for Serosurveillance date = 2004-09-17 pages = extension = .txt mime = text/plain words = 2800 sentences = 133 flesch = 44 summary = A peptide-based enzyme-linked immunosorbent assay (ELISA) can be used for retrospective serosurveillance of severe acute respiratory syndrome (SARS) by helping identify undetected chains of disease transmission. A peptide-based enzyme-linked immunosorbent assay (ELISA) can be used for retrospective serosurveillance of severe acute respiratory syndrome (SARS) by helping identify undetected chains of disease transmission. Such surveillance may be key to tracking the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) because mild and asymptomatic cases of SARS-CoV infection that do not meet the World Health Organization's case definition (1) have been identified by immunoassays (2) (3) (4) , and SARS-CoV-like viruses have been isolated from wild mammals (5) . The diagnostic sensitivity of the peptide ELISA was 100% on a panel of 69 convalescent-phase serum samples from SARS patients provided as a reference panel by the Center for Disease Control, Department of Health, Taiwan. The peptide ELISA was evaluated for specificity on serum samples drawn from patients associated with typical and atypical respiratory pathogens other than SARS-CoV (National Taiwan University Hospital). cache = ./cache/cord-321691-46la29tm.txt txt = ./txt/cord-321691-46la29tm.txt === reduce.pl bib === id = cord-334896-3g75spkc author = MINAMI, Shohei title = Establishment of serological test to detect antibody against ferret coronavirus date = 2016-03-03 pages = extension = .txt mime = text/plain words = 2493 sentences = 150 flesch = 59 summary = Since there is no available serological methods to detect antibodies to ferret coronavirus (FRCoV), an enzyme-linked immunosorbent assay (ELISA) using recombinant partial nucleocapsid (N) proteins of the ferret coronavirus (FRCoV) Yamaguchi-1 strain was developed to establish a serological method for detection of FRCoV infection. This different reactivity was also confirmed by immunoblot analysis using the serum from the ferret.Therefore, the a.a. 1–179 of the N protein was used as an ELISA antigen. In this study, an enzyme-linked immunosorbent assay (ELISA) using recombinant N proteins was established and applied to investigate the seroprevalence of FRcoV infection in Japan. The plasma of No.10 and serum of ferret No.22 showed differ-ent reactivities from the other samples in ELISA (Fig. 2) . In this study, we attempted to clarify the seroprevalence of FRcoV in Japan and developed an ELISA using two Yamaguchi-1 strain recombinant N proteins, GST-N (1-179) and GST-N (180-374). cache = ./cache/cord-334896-3g75spkc.txt txt = ./txt/cord-334896-3g75spkc.txt === reduce.pl bib === id = cord-336899-zngp67tb author = Kadkhoda, Kamran title = COVID‐19: are neutralizing antibodies neutralizing enough? date = 2020-06-03 pages = extension = .txt mime = text/plain words = 1211 sentences = 73 flesch = 51 summary = The use of convalescent-phase plasma (CP) in severely ill patients with COVID-19 has been attempted on an individual basis 1 and it is being used in the context of ongoing clinical trials; thus, it is pivotal to understand the potential risks and caveats of using CP particularly taking immunopathologic phenomena into account. In this context, previous exposure to common coronaviruses would lead to an early and high-titer immune response to SARS-CoV-2. The donors had neutralizing antibody titers of more than 640 at the time of donation while severely ill patients had relatively similar titers before transfusion as high as 640 (range, 160-640; GMT, 367). 7 The US Food and Drug Administration currently recommends using donated plasma with neutralizing antibody titers of 160 or at a minimum a titer of 80 in severely ill patients. cache = ./cache/cord-336899-zngp67tb.txt txt = ./txt/cord-336899-zngp67tb.txt === reduce.pl bib === id = cord-331277-fjsuo3yy author = Hoste, Alexis C.R. title = Two serological approaches for detection of antibodies to SARS-CoV-2 in different scenarios: A screening tool and a point-of-care test date = 2020-08-11 pages = extension = .txt mime = text/plain words = 2407 sentences = 136 flesch = 52 summary = Two serological tools based on a Double Recognition assay (Enzyme-Linked Immunosorbent Assay, DR-ELISA and Lateral Flow Assay, DR-LFA) to detect total antibodies to SARS-CoV-2, have been developed based on the recombinant nucleocapsid protein. Therefore, the aim of the present work was the development of serological tools to determine the presence of antibodies against SARS-CoV-2 in the population, as an indicator of an ongoing or previous infection. In the current study, a Double Recognition Enzyme-Linked Immunosorbent Assay (DR-ELISA) was developed to determine the presence of immunoglobulins of different classes (IgG, IgM and IgA) to SARS-CoV-2 in human serum, to support the diagnosis of COVID-19. In the present study, we developed two serological assays using the recombinant N protein Table 1, a group of 14 serum samples from early days post infection, positive to COVID-19 by respiratory-PCR yet still negative in the commercial serological assay (with seroconversion a few days later) were also tested in our assays. cache = ./cache/cord-331277-fjsuo3yy.txt txt = ./txt/cord-331277-fjsuo3yy.txt === reduce.pl bib === id = cord-335121-ro3x3qa3 author = Ingram, George A. title = A comparative assessment of four serological methods used in the detection and measurement of anti-parasite antibodies in the serum of the amphibian, bufo viridis date = 1988-04-30 pages = extension = .txt mime = text/plain words = 3116 sentences = 180 flesch = 50 summary = Abstract Antibodies against Crithidia fasciculata choanomastigotes were detected in green toad (Bufo viridis) sera by direct agglutination, indirect haemagglutination (IHA), complement-fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA), Correlation coefficients (r) were calculated for comparisons between each of the techniques and regression formulae derived in order to convert antibody levels as determined by one immunological method to that of another. In this paper we present the results of acomparative assessment of four serological tests (direct agglutination, indirect haemagglutination, complement-fixation test and ELISA) used to detect and to determine the levels of antibodies in the sera of green toads (B. fasciculata and the number of control and immune sera containing detectable antibodies were the lowest for DA and CFT, intermediate for IHA and highest for the ELISA method (Table 1) . Nevertheless the method of antigen preparation ma> not be a salient criterion for antibody estimation in immune sera because correlation values of over 89% (f'< 0.001) were found when IHA titres were compared to those of ELISA. cache = ./cache/cord-335121-ro3x3qa3.txt txt = ./txt/cord-335121-ro3x3qa3.txt === reduce.pl bib === id = cord-335591-r0x8yaqj author = Ohnishi, Kazuo title = Establishment and Characterization of Monoclonal Antibodies Against SARS Coronavirus date = 2007-11-28 pages = extension = .txt mime = text/plain words = 3126 sentences = 242 flesch = 67 summary = The hybridomas produce monoclonal antibodies that recognize viral component molecules, including the spike protein (S) and the nucleocapsid protein (N), enabling the immunological detection of SARS-CoV by immunofluorescence staining, immunoblot, or an antigen-capture ELISA system. Based on clinical experience, several options have been considered in the quest to develop the capacity to accurately diagnose SARS-CoV infection, including molecular biology techniques and serological tests such as antigen-capture ELISA assay and immunofluorescence assay to detect virus-infected cells in respiratory swabs (3-7) . These mAbs enable the general immunological detection of SARS-CoV by methods such as immunofluorescent staining, immunoblotting, and immunohistology, in addition to the construction of a highly sensitive antigen-capture sandwich ELISA (6). The UV-inactivated purified SARS-CoV samples (see Note 1), which are serially diluted with 1% OVA/PBS-Tween, are added to the wells and incubated for 1 h at room temperature cache = ./cache/cord-335591-r0x8yaqj.txt txt = ./txt/cord-335591-r0x8yaqj.txt === reduce.pl bib === id = cord-322529-3xn5v54s author = Rodák, L. title = Verification of Sensitivity and Specificity of Group A Rotavirus Detection in Piglets Faeces with Monoclonal Blocking ELISA Methods date = 2004-06-30 pages = extension = .txt mime = text/plain words = 3215 sentences = 196 flesch = 48 summary = title: Verification of Sensitivity and Specificity of Group A Rotavirus Detection in Piglets Faeces with Monoclonal Blocking ELISA Methods Selected competitive blocking ELISA (CB‐ELISA) and electron microscopy (EM) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. The sensitivity and specificity of the CB‐ELISA was verified both by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhoea virus (PEDV) in each analysis and by comparative examination of samples with the commercial ELISA kit. Sensitivity comparison of three variants of the blocking ELISA method of rotavirus A detection were performed by box titrations in microtitre plate wells pre-coated with binding antibodies. Sensitivity of the CB-ELISA method and DAS-ELISA kit was compared by examination of faecal sample of experimentally infected piglet twofold diluted 2· to 1024·. By examination of positive faecal sample twofold diluted 2· to 1024·, at least 10 times higher sensitivity of CB-ELISA method was demonstrated (Table 2) . cache = ./cache/cord-322529-3xn5v54s.txt txt = ./txt/cord-322529-3xn5v54s.txt === reduce.pl bib === id = cord-351952-lhhjax3s author = Pickering, Suzanne title = Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date = 2020-09-24 pages = extension = .txt mime = text/plain words = 5310 sentences = 247 flesch = 48 summary = Highly specific in-house ELISAs were developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays—a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs)—on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. Accordingly, we developed a highly specific semi-quantitative ELISA for the detection of anti-spike (S), -S receptor binding domain (RBD) and -N antibodies, and used this to cross-evaluate ten commercial antibody tests (seven lateral flow immunoassays (LFIAs), one chemiluminescent assay and two ELISAs) on a collection of 110 serum samples from confirmed RNA positive patients, and 50 pre-pandemic samples from March 2019. With no existing standardised diagnostic test for the assessment of the serological response to SARS-CoV-2, we started by comparing commercial serological assays with the configuration of the in-house ELISA most likely to represent antibodies detected by the commercial tests (detection of anti-S IgM and IgG antibodies), and that had high specificity and sensitivity (S1 Table) . cache = ./cache/cord-351952-lhhjax3s.txt txt = ./txt/cord-351952-lhhjax3s.txt === reduce.pl bib === id = cord-347374-mryazbnq author = Okba, Nisreen M.A. title = Severe Acute Respiratory Syndrome Coronavirus 2−Specific Antibody Responses in Coronavirus Disease Patients date = 2020-07-17 pages = extension = .txt mime = text/plain words = 3565 sentences = 186 flesch = 51 summary = Using serum samples from patients with PCR-confirmed SARS-CoV-2 infections, other coronaviruses, or other respiratory pathogenic infections, we validated and tested various antigens in different in-house and commercial ELISAs. We demonstrated that most PCR-confirmed SARS-CoV-2–infected persons seroconverted by 2 weeks after disease onset. Using a well-characterized cohort of serum samples from PCR-confirmed SARS-CoV-2 and patients PCR-confirmed to be infected with seasonal coronaviruses and other respiratory pathogens, we validated and tested various antigens in different platforms developed in-house, as well as a commercial platform. We evaluated SARS-CoV-2-specific antibody responses in severe and mild cases by using serum samples collected at different times postonset of disease from 3 PCR-confirmed COVID-19 patients from France. We tested serum samples for SARS-CoV-2specific antibodies by using different ELISAs. After infection, all 3 patients seroconverted between days 13 and 21 after onset of disease (Figure 1) , and antibodies were elicited against the SARS-CoV-2 S, S1 subunit, and RBD, but only 2/3 patients had detectable antibodies to the N-terminal (S1 A ) domain. cache = ./cache/cord-347374-mryazbnq.txt txt = ./txt/cord-347374-mryazbnq.txt === reduce.pl bib === id = cord-342024-kaku49xd author = Espejo, Andrea P title = Review of Current Advances in Serologic Testing for COVID-19 date = 2020-06-25 pages = extension = .txt mime = text/plain words = 6480 sentences = 373 flesch = 50 summary = • The use of total antibody or simultaneous IgG/IgM measurements (regardless of method) significantly adds sensitivity to reverse transcription polymerase chain reaction testing protocols early post onset of symptoms and becomes the most accurate diagnostic test at later time points. The SP, RBD, and NP proteins appear to be the main targets of the humoral immune response in coronavirus infections including SARS-CoV-2 and were the antigens used in the majority of the serologic assays examined in this literature review. Overall, while these results are similar to that reported in a review of serologic testing for MERS-CoV and SARS-CoV, they may have been affected by choice of target antigens, the various immunoassay kits, and the level of detail of case history used to categorize the time of sample acquisition post onset of symptoms. cache = ./cache/cord-342024-kaku49xd.txt txt = ./txt/cord-342024-kaku49xd.txt === reduce.pl bib === id = cord-346574-u28y1ttw author = Chen, Keyan title = Development and evaluation of an immunochromatographic strip for rapid detection of porcine hemagglutinating encephalomyelitis virus date = 2012-08-24 pages = extension = .txt mime = text/plain words = 5509 sentences = 277 flesch = 50 summary = At present, various laboratory methods are available for the detection and surveillance of PHE-CoV, including virus isolation [1] , hemagglutination/hemagglutination inhibition (HA/HI) tests [13] , immunohistochemistry (IHC) assays [10] , and molecular tools such as nestedpolymerase chain reaction (nested PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR) that enable detection of specific CoV RNA sequences from infected tissues [14, 15] . (1) (2) (3) In this study, an immunochromatographic strip with high sensitivity and specificity was developed for the detection of PHE-CoV, combining monoclonal antibody (MAb) and colloidal gold immunochromatography (GICA), and the resulting product is suitable for the surveillance of PHE-CoV. Thus, a lot of brain tissue samples were collected from deceased piglets with suspected PHE-CoV infection, and using RT-PCR and ELISA as reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. cache = ./cache/cord-346574-u28y1ttw.txt txt = ./txt/cord-346574-u28y1ttw.txt === reduce.pl bib === id = cord-022650-phsr10jp author = nan title = Abstracts TPS date = 2018-08-14 pages = extension = .txt mime = text/plain words = 119675 sentences = 7010 flesch = 55 summary = 0685 | Skin prick test reactivity to aeroallergens in adult allergy clinic in a tertiary hospital: a 12-year retrospective study Results: Five different human sera were screened for specific IgE level against 29 different allergen sources using test methods of three different suppliers. Conclusion: This multicenter prospective study confirmed that stepwise single-dose OFC to egg will help to clarify the severity of egg allergy, and will contribute to improved food allergy manageMethod: The study design was a retrospective cohort study extracting data from the electronic chart of children older than 4 years who visited our out-patient clinic for egg or milk allergy and who underwent an oral food challenge test (OFC) twice within 24 months between November 2013 and December 2017. Results: In the base case analysis, using Italy clinical practice patients with moderate-to severe allergic rhino-conjunctivitis (SS ranging from 6 to 15 points) and a mean age at entry of 21 years, both SCIT and SLIT were associated with increased cost but superior efficacy compared to pharmacotherapy alone. cache = ./cache/cord-022650-phsr10jp.txt txt = ./txt/cord-022650-phsr10jp.txt === reduce.pl bib === id = cord-344871-486sk4wc author = Wu, Jianping title = Biochemical and structural characterization of the interface mediating interaction between the influenza A virus non-structural protein-1 and a monoclonal antibody date = 2016-09-16 pages = extension = .txt mime = text/plain words = 6992 sentences = 379 flesch = 58 summary = We have previously shown that a non-structural protein 1 (NS1)-binding monoclonal antibody, termed as 2H6, can significantly reduce influenza A virus (IAV) replication when expressed intracellularly. As comparative ELISA in this and previous studies 29 showed that residues N48 and T49 in NS1(RBD) are important for the interaction with mAb 2H6, they were defined as active residues involved in the binding interaction to generate a series of models of the NS1(RBD) and 2H6-Fab complex. Overall, the predicted model from cluster 2 is consistent with our comparative ELISA data and suggests that residues N48 and T49 are important for the binding between NS1(RBD) and 2H6-Fab because their side-chains could make hydrogen bonds with residues in the VH-CDR2 of the Fab. In addition, R44 of NS1(RBD) was distal from the antibody-antigen interface, which is consistent with the results from comparative ELISA ( Figure S1 ) showing that substitution of R44 of NS1(RBD) with K did not affect its interaction with mAb 2H6. cache = ./cache/cord-344871-486sk4wc.txt txt = ./txt/cord-344871-486sk4wc.txt === reduce.pl bib === id = cord-348660-qnbgywgy author = Yilmaz, Huseyin title = Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen date = 2018-07-09 pages = extension = .txt mime = text/plain words = 3921 sentences = 183 flesch = 43 summary = Following optimization of the ELISA protocol, 18 test sera were obtained from broiler chickens exposed to natural wild-type IBV infection, 18 test sera from broiler chickens vaccinated with a live-attenuated commercial IBV vaccine, and sera obtained at different time-points from chicks immunized with recombinant IBV N protein (described above) were analyzed to detect IBV N-specific antibodies. To assess the reliability of the performance of our in-house indirect IBV N ELISA, a panel of sera was obtained from chickens naturally infected with local wild-type IBV strains, chickens vaccinated with live-attenuated commercial IBV vaccine, and chickens immunized with recombinant IBV N protein (expressed in a baculovirus expression system). This represents the first study in Turkey that expressed recombinant IBV N protein in baculovirus and examined its reactivity against antisera obtained from Turkish chickens for potential use as antigen Fig. 5 Detection of IBV N specific antibodies in sera obtained from naturally infected chicken (a) and vaccinated chickens (b) using an in-house IBV-N ELISA and a commercial ELISA. cache = ./cache/cord-348660-qnbgywgy.txt txt = ./txt/cord-348660-qnbgywgy.txt === reduce.pl bib === id = cord-349744-8cg5yj20 author = Lassaunière, Ria title = Evaluation of nine commercial SARS-CoV-2 immunoassays date = 2020-04-10 pages = extension = .txt mime = text/plain words = 2649 sentences = 145 flesch = 53 summary = The results showed 100% specificity for the Wantai SARS-CoV-2 Total Antibody ELISA, 93% for the Euroimmun IgA ELISA, and 96% for the Euroimmun IgG ELISA with sensitivities of 90%, 90%, and 65%, respectively. While the four POC tests evaluated according to illness duration were often weakly positive or detected only IgG or IgM during the early phase (data not shown), their sensitivities were comparable to the Wantai Total Ab ELISA and Euroimmun IgA ELISA in all three phases. In the present study, three SARS-CoV-2-specific commercial ELISA assays and six POC rapid tests were evaluated using sera from hospitalized adult patients with PCR-confirmed diagnoses for SARS-CoV-2 and a collection of control serum samples taken before the emergence of the virus in China in December 2019. Overall, the Wantai Total Ab ELISA had superior sensitivity and specificity compared to both Euroimmun IgA and IgG ELISAs. The POC tests varied notably, with the best performance observed for the test produced by AutoBio Diagnostics, followed by the tests produced by Dynamiker Biotechnology and CTK Biotech. cache = ./cache/cord-349744-8cg5yj20.txt txt = ./txt/cord-349744-8cg5yj20.txt === reduce.pl bib === id = cord-344581-h7ikjgic author = Ong, David S.Y. title = Comparison of diagnostic accuracies of rapid serological tests and ELISA to molecular diagnostics in patients with suspected COVID-19 presenting to the hospital date = 2020-06-02 pages = extension = .txt mime = text/plain words = 2071 sentences = 120 flesch = 54 summary = OBJECTIVES: To assess the diagnostic performance of rapid lateral flow immunochromatographic assays (LFAs) compared to an enzyme-linked immunosorbent assay (ELISA) and nucleic acid amplification tests (NATs) in suspected coronavirus disease 2019 (COVID-19) patients. In the total cohort, Orient Gene Biotech COVID-19 IgG/IgM Rapid Test LFA had a sensitivity of 43/99 (43%; 95% CI 34-53) and specificity of 126/129 (98%; 95% CI 95-100). CONCLUSIONS: There is large variability in diagnostic test performance between rapid LFAs, but overall limited sensitivity and high specificity in acutely admitted patients. First, in a pilot phase 20 NAT-positive and 5 NAT-negative patients were retrospectively selected for which six LFAs were performed on heparin plasma samples obtained upon hospital presentation ( Figure S1 ), which corresponded to the dates of molecular testing. This study shows that the sensitivity of LFA was low in patients suspected for COVID-19 presenting to the hospital, but it improved in patients with at least seven days of symptoms and in those with CRP levels >100 mg/L upon presentation. cache = ./cache/cord-344581-h7ikjgic.txt txt = ./txt/cord-344581-h7ikjgic.txt === reduce.pl bib === id = cord-346320-ysgz6adr author = Arabi, Yaseen M. title = Feasibility of Using Convalescent Plasma Immunotherapy for MERS-CoV Infection, Saudi Arabia date = 2016-09-17 pages = extension = .txt mime = text/plain words = 4239 sentences = 203 flesch = 49 summary = We explored the feasibility of collecting convalescent plasma for passive immunotherapy of Middle East respiratory syndrome coronavirus (MERS-CoV) infection by using ELISA to screen serum samples from 443 potential plasma donors: 196 patients with suspected or laboratory-confirmed MERS-CoV infection, 230 healthcare workers, and 17 household contacts exposed to MERS-CoV. We explored the feasibility of collecting convalescent plasma for passive immunotherapy of Middle East respiratory syndrome coronavirus (MERS-CoV) infection by using ELI-SA to screen serum samples from 443 potential plasma donors: 196 patients with suspected or laboratory-confirmed MERS-CoV infection, 230 healthcare workers, and 17 household contacts exposed to MERS-CoV. In collaboration with the King Abdullah International Medical Research Center, the Gulf Cooperation Council Infection Control Center, and the World Health Organization (WHO)-International Severe Acute Respiratory and Emerging Infection Consortium MERS-CoV Working Group, we developed a study protocol to screen potential donors, collect high-titer convalescent plasma, and administer the plasma in a clinical trial (13) . cache = ./cache/cord-346320-ysgz6adr.txt txt = ./txt/cord-346320-ysgz6adr.txt === reduce.pl bib === id = cord-340960-abanr641 author = Brigger, D. title = Accuracy of serological testing for SARS‐CoV‐2 antibodies: first results of a large mixed‐method evaluation study date = 2020-09-30 pages = extension = .txt mime = text/plain words = 4479 sentences = 289 flesch = 50 summary = In a mixed‐design evaluation study, we compared the diagnostic accuracy of serological immunoassays that are based on various SARS‐CoV‐2 proteins and assessed the neutralizing activity of antibodies in patient sera. A total of 54 randomly selected sera from individuals who were tested positive in either of the three ELISA immunoassays as well as 6 negative controls were assessed in a live SARS-CoV-2 neutralization assay (all collected in April 2020). Recombinantly expressed RBD has been used to establish an in-house ELISA for the detection of IgM and IgG anti-SARS-CoV-2 antibodies in human serum samples (supplementary Fig. 1a,b) . A total of 54 randomly selected sera from individuals who were tested positive in either of the three ELISA immunoassays as well as 6 negative controls were assessed in a live SARS-CoV-2 neutralization assay using ACE2-expressing Vero-E6 cells (34 inpatient samples, and 26 samples of medical personnel). cache = ./cache/cord-340960-abanr641.txt txt = ./txt/cord-340960-abanr641.txt === reduce.pl bib === id = cord-351498-bmq6zcb0 author = Martínez-Sernández, Victoria title = Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis date = 2018-03-21 pages = extension = .txt mime = text/plain words = 8277 sentences = 401 flesch = 48 summary = In the present study, we developed and tested three indirect ELISAs using rFhpCL1, rFhpCL2, and rFhpCL5 and evaluated their recognition by sera from sheep and cattle naturally infected with F. In the present study, we developed and tested indirect ELISAs based on recombinant procathepsin L1 (rFhpCL1), rFhpCL2, or rFhpCL5, in order to investigate which of these targets are best recognized by sera from sheep and cattle naturally infected with F. Finally, the r values obtained on comparing the four ELISA methods for Fasciola-infected cattle and sheep sera are shown in Table 3 . These results suggest that the use of a single recombinant cathepsin/ procathepsin as target antigen in ELISAs for serodiagnosis of fascioliasis may limit the sensitivity of the assay when testing sera from some species, particularly cattle. (2001) who tested sera from sheep and cattle harboring other parasites, mainly nematodes, suggesting that the cross-reactivity may be due to common epitopes between recombinant Fasciola cathepsins and antigens present in other parasites. cache = ./cache/cord-351498-bmq6zcb0.txt txt = ./txt/cord-351498-bmq6zcb0.txt === reduce.pl bib === id = cord-348161-757c51xw author = Petrosova, A. title = Development of a highly sensitive, field operable biosensor for serological studies of Ebola virus in central Africa date = 2007-03-26 pages = extension = .txt mime = text/plain words = 5428 sentences = 264 flesch = 48 summary = We employed a photo immobilization methodology based on a photoactivatable electrogenerated poly(pyrrole-benzophenone) film deposited upon an indium tin oxide (ITO) modified conductive surface fiber-optic. The photochemically modified optical fibers were tested as an immunosensor for detection of antibodies against Ebola virus, in animal and human sera, by use of a coupled chemiluminescent reaction. In this study we present a newly developed optical immunosensor for detection of antibodies to Ebola virus strains Zaire and Sudan, by using a photoimmobilization methodology based on a photoactivable electrogenerated polymer film. The optical fibers coated with poly(pyrrole-benzophenone) were soaked in diluted solution containing inactivated Ebola virus antigen (approximately 7.5 g/ml, the concentration was determined by Micro BCA Protein assay kit, PIERCE) and irradiated with UV light. The calibration curve obtained from the optical fiber immunosensor and ELISA for the detection of anti-Ebola subtype Zaire antibodies. cache = ./cache/cord-348161-757c51xw.txt txt = ./txt/cord-348161-757c51xw.txt === reduce.pl bib === id = cord-353614-z4fpy607 author = Kusano, Nario title = Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies date = 2007-02-16 pages = extension = .txt mime = text/plain words = 2976 sentences = 147 flesch = 54 summary = title: Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies A simple and rapid immunochromatographic assay (ICA) to detect Satsuma dwarf virus (SDV) was developed using colloidal gold conjugates of anti-SDV monoclonal antibodies. Comparison of the sensitivities of ICA and DAS-ELISA Out of the three combinations of using three Mabs for ICA, clone 2G2 and clone 4E6, which had a different paratope group (PG), were more sensitive and suitable to the colloidal gold conjugate and coating antibody on the nitrocellulose strip for detection of SDV, respectively (data not shown). Suspensions of colloidal gold conjugate at various concentrations were applied to the reagent pad, and ICA The immunochromatographic devices were prepared under the following conditions: Case 3-1, 1 mg/ml of anti-SDV polyclonal antibody was deposited onto a nitrocellulose membrane to make a test line. cache = ./cache/cord-353614-z4fpy607.txt txt = ./txt/cord-353614-z4fpy607.txt === reduce.pl bib === id = cord-353190-7qcoxl81 author = Nicklas, Werner title = Viral Infections of Laboratory Mice date = 2012-05-17 pages = extension = .txt mime = text/plain words = 27775 sentences = 1482 flesch = 39 summary = This chapter covers infections of mice with the following viruses: herpesviruses, mousepox virus, murine adenoviruses, polyomaviruses, parvoviruses, lactate dehydrogenase-elevating virus, lymphocytic choriomeningitis virus, mammalian orthoreovirus serotype 3, murine hepatitis virus, murine norovirus, murine pneumonia virus, murine rotavirus, Sendai virus, and Theiler's murine encephalomyelitis virus. These results are very difficult to summarize because the outcome of experimental infection in laboratory mice depends on various factors such as mouse strain and age, virus strain and passage history [26] , virus dose and route of inoculation [24] . Experimental infection of laboratory mice with MHV-68 is a frequently used model system for the study of human gammaherpesvirus pathogenesis, e.g. of Kaposi's sarcoma-associated herpesvirus or Epstein-Barr virus (EBV) [62, 63] which are members of the same subfamily. Early descriptions of naturally occurring disease may have been complicated by concurrent infections such as MHV (murine hepatitis virus) or murine rotavirus A (MuRV-A)/epizootic diarrhoea of infant mice (EDIM) virus that contributed to the severity of the lesions especially in liver, pancreas, CNS and intestine. cache = ./cache/cord-353190-7qcoxl81.txt txt = ./txt/cord-353190-7qcoxl81.txt === reduce.pl bib === id = cord-019347-tj3ye1mx author = nan title = ABSTRACT BOOK date = 2010-02-19 pages = extension = .txt mime = text/plain words = 107926 sentences = 6940 flesch = 53 summary = Method:Case Report:A 15y/o w/f athlete presented with a two month history of recurrent hives and angioedema which she associated with ingestion of Halloween candy .One week before evaluation she had hives with Coconut as well.Her history was othewise unremarkable except for recurrent UTI'S, annual sinusitis, pneumonia in 1998 as well as migraines.She denied sexual activity.Her physical exam was normal.Results:An evaluation for autoimmune disease revealed normal ESR, ANA, DSDNA, mono and hepatitis serology as well as lyme titers however her CH50 was low17u/ml(normal 26-58U/ml)and evaluation of complement revealed c4 14mg/dl(normal 16-47mg//dl)and c2 <1.3mg/dl(normal 1.6-3.5mg/dl)with normal c3, c5-c9.Her father had nor-malc4 but c2 was 1.4mg/dl (normal 1.6-3.5mg/dl)Her sister had c2 of 1.5mg/dl and normal c4 and her mother had normal c2 and c4.Her workup included positive prick skin test to ragweed, ash and grass and she was started on Rhinocort and Clarinex seasonally.She has been followed for one year with resolution of hives and is asymptomatic.Her diagnosis had been confirmed by a pediatric rheumatologist.Conclusion;We present an atypical case of C2 complement deficiency in an currently asymptomatic individual. cache = ./cache/cord-019347-tj3ye1mx.txt txt = ./txt/cord-019347-tj3ye1mx.txt === reduce.pl bib === id = cord-022501-9wnmdvg5 author = nan title = P1460 – P1884 date = 2015-12-28 pages = extension = .txt mime = text/plain words = 128256 sentences = 7808 flesch = 51 summary = Methods: Using published data on (1) the prevalence of MRSA and other bacterial pathogens causing cSSSI in the US, (2) the in-vitro susceptibility rates of commonly used regimens in cSSSI in the US in relation to the most pervasive pathogens identified above, and (3) estimated costs of failure of initial, empiric treatment from a recent study of a large US multi-hospital database, we developed a model to predict the expected clinical and economic impact of increasing prevalence of MRSA. Small outbreaks of VEB-1 ESBL producing Acinetobacter baumannii in Belgian nursing homes and hospitals through cross-border transfer of patients from northern France Methods: From 01/04 to 03/05, all Belgian acute hospitals were invited to report cases of nosocomial infections/colonisations due to MDR Ab isolates presenting a resistance profile similar to the French epidemic strain (resistance to all agents except carbapenems and colistin) and to send such isolates to the reference laboratory for phenotypic confirmation and for genotypic characterization (PCR of VEB-1 and class 1 Integron, PFGE typing). cache = ./cache/cord-022501-9wnmdvg5.txt txt = ./txt/cord-022501-9wnmdvg5.txt === reduce.pl bib === id = cord-022888-dnsdg04n author = nan title = Poster Sessions date = 2009-08-19 pages = extension = .txt mime = text/plain words = 188640 sentences = 9313 flesch = 45 summary = Methods: Phospho-specific Western blot analyses were performed to verify the functionality of the different IFN-g pathway components, intra-and extracellular flow cytometry experiments were employed to determine the expression of antigen processing components and HLA class I cell surface antigens, quantitative real time-PCR experiments to confirm the absence of JAK2 and presence of pathway relevant molecules as well as, genomic PCR and chromosome typing technique to prove the deletion of JAK2. In order to accomplish these objectives we induced priming or tolerance of ovalbumin (OVA 323-339 peptide)-specific T cells from DO11.10 TCR transgenic mice in vitro or, following adoptive transfer of near physiologically relevant numbers of such cells into recipients, in vivo and correlated functional outcome (via proliferation and cytokine readout assays or antibody production) with E3 ubiquitin-protein ligases expression and the ubiquitination status of the TCR signalling machinery. cache = ./cache/cord-022888-dnsdg04n.txt txt = ./txt/cord-022888-dnsdg04n.txt === reduce.pl bib === id = cord-023095-4dannjjm author = nan title = Research Abstract Program of the 2011 ACVIM Forum Denver, Colorado, June 15–18, 2011 date = 2011-05-03 pages = extension = .txt mime = text/plain words = 134226 sentences = 6834 flesch = 51 summary = The purpose of this study was to determine the short-term effects of ivabradine on heart rate (HR), blood pressure, left ventricular (LV) systolic and diastolic function, left atrial (LA) performance, and clinical tolerance in healthy cats after repeated oral doses. The goal of this study was to investigate the relationship between heart rate and ECG time intervals to body mass in apparently healthy horses and ponies and to calculate normal ranges for different weight groups. This study aimed to investigate the prevalence of hypercoagulability in PLN dogs based on thromboelastography (TEG), and to determine whether hypercoagulability in these patients could be predicted by clinical assessments that identify systemic hypertension (systolic blood pressure 4 160 mmHg), hypoalbuminemia (serum albumin o 2.7 mg/dl), antithrombin activity (o 70%), and degree of proteinuria (urine protein:creatinine ratio [UPC] ! cache = ./cache/cord-023095-4dannjjm.txt txt = ./txt/cord-023095-4dannjjm.txt === reduce.pl bib === id = cord-015021-pol2qm74 author = nan title = Third International Congress on the Immune Consequences of Trauma, Shock and Sepsis —Mechanisms and Therapeutic Approaches date = 1994 pages = extension = .txt mime = text/plain words = 162327 sentences = 9379 flesch = 50 summary = It is our current understanding that LPS is responsible for many of the pathophysiological events observed during gramnegative infections and that one of the major mechanisms leading to shock and death is the LPS-induced activation of macrophages resulting in the production and release of lipid and peptide mediators, among which tumor necrosis factor seems to be the most important. However plasma IL-6 estimation revealed a statistically significant reduction at 6 hours in tanrine-treated animals compared to glycino and TW controls ( Objective: To evaluate the effects of allogeneic blood transfusion, thermal injury and bacterial garage on interteukin 4 (IL-4), tumor necrosis factor alpha (TNF) production and host mortality and to study if the administration of thymopentth (THY) could affect these events. cache = ./cache/cord-015021-pol2qm74.txt txt = ./txt/cord-015021-pol2qm74.txt === reduce.pl bib === id = cord-031907-ilhr3iu5 author = nan title = ISEV2020 Abstract Book date = 2020-07-15 pages = extension = .txt mime = text/plain words = 200999 sentences = 11528 flesch = 44 summary = L.M., and the National Institutes of Health (R35GM119623) to T.R.G. The addition of a size exclusion chromatography step to various urinary extracellular vesicle concentrating methods reveals differences in the small RNA profile Introduction: Urinary extracellular vesicles (EVs) and their RNA cargo are a novel source of biomarkers for various diseases, however non-vesicular RNA (e.g. associated with proteins) is also present within urine. We then evaluated efficiency of heart targeting for eAAV9 or eAAV6 and standard AAV9 or AAV6 encoding for EGFP, mCherry or firefly luciferase in different human cell lines in vitro, in black mouse and in passive immunity nude mouse model in vivo using flow cytometry, confocal microscopy, Langendorff perfusion system and Methods: HLHS patients (n = 3) after Glenn procedure and swine (n = 3) after PAB were given RV injections of allogeneic/xenogeneic MSCs. Donor-specific, HLA-I+, exosomes were isolated from plasma. cache = ./cache/cord-031907-ilhr3iu5.txt txt = ./txt/cord-031907-ilhr3iu5.txt === reduce.pl bib === id = cord-022940-atbjwpo5 author = nan title = Poster Sessions date = 2016-09-07 pages = extension = .txt mime = text/plain words = 241182 sentences = 12746 flesch = 47 summary = We have studied the effect of inhibition of IRE1 (inositol requiring enzyme 1), which is a central mediator of endoplasmic reticulum stress and controls cell proliferation and tumor growth, on hypoxic regulation of the expression of different proliferation related genes in U87 glioma cells. Transient inhibition of Akt and mTOR protein kinase activation in tumor cells followed by reactivation of signaling pathway did not result in a time-dependent difference on EGFR, HER2 and HER3 expression levels. In our study we aimed to determine cytotoxic effect of RES in K562 human CML cell line and to evaluate the expressions of miRNAs that are associated with genetics of leukemia after treatment with RES; to investigate target genes of miRNAs which show significant expression alterations and molecular mechanisms of RES treatment. cache = ./cache/cord-022940-atbjwpo5.txt txt = ./txt/cord-022940-atbjwpo5.txt === reduce.pl bib === id = cord-015394-uj7fe5y6 author = nan title = Scientific Abstracts date = 2008-12-23 pages = extension = .txt mime = text/plain words = 242330 sentences = 15267 flesch = 52 summary = Studies involving immunohistochemical analysis of normal ovaries have shown that granulosa cells express significantly higher levels of the activator protein-1 (AP-1) transcription factor, cFos compared to theca cells, where cFos expression is virtually absent. Following acute hypoxia (0.5% O2) for one to six hours, RhoA mRNA, total protein and activation (RhoA-GTP) levels were analysed, using semi-quantitative PCRs and western blot, and compared to normoxic non-pregnant human uterine smooth muscle control cells. Since there is an urgent need for non-invasive methods for determination of fetal (F) and placental (P) function, this study was designed to evaluate the genes differently and commonly expressed in P tissue and leukocytes in maternal (M) and F circulation.Material and Methods. The current study: 1) localized IL-6 mRNA levels in preeclamptic versus normal decidual sections; 2) evaluated mechanisms regulating IL-6 synthesis by targeting intracellular signaling pathways with specific inhibitors; 3) identified potential IL-6 targets by immunolocalizing the IL-6 receptor (IL-6R) to specific cell types in placental bed biopsies. cache = ./cache/cord-015394-uj7fe5y6.txt txt = ./txt/cord-015394-uj7fe5y6.txt === reduce.pl bib === id = cord-344309-6c2wttxg author = Lin, Huixing title = Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein date = 2018-08-20 pages = extension = .txt mime = text/plain words = 4411 sentences = 221 flesch = 55 summary = title: Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein In this study, an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant truncated spike (S) protein of PEDV was developed and validated. This indirect ELISA was compared to indirect immunoinfluscent assay (IFA), and the overall coincidence rate was 96.74% based on testing 368 clinical serum samples with different PEDV antibody levels. Finally, the S1 indirect ELISA was applied to detect serum antibodies of 3304 field samples collected from different pig farms in eastern China, and it presented an overall substantial agreement on the PEDV infection status. Therefore, this study selected a gene fragment within the S1 subunit as a coating antigen to develop an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of PEDV antibodies. Detection of antibodies against porcine epidemic diarrhea virus in serum and colostrum by indirect ELISA cache = ./cache/cord-344309-6c2wttxg.txt txt = ./txt/cord-344309-6c2wttxg.txt ===== Reducing email addresses cord-011212-ovjdzyxv cord-015147-h0o0yqv8 cord-334165-7gfk554m cord-351498-bmq6zcb0 Creating transaction Updating adr table ===== Reducing keywords cord-000434-ff2zadol cord-003208-lwirkob3 cord-004101-0r2g5p1i cord-007480-grndfx7b cord-003471-tr3ageky cord-003315-r1wkx0ml cord-003656-7mzsaz7a cord-009922-t1hoox6e cord-007497-nn5l5rai cord-011212-ovjdzyxv cord-007495-gpz4gkv3 cord-003623-n01rgqyv cord-001446-mpuovmeb cord-012891-heqsfzkm cord-003859-k8wfyj9b cord-007644-7bsixsgd cord-014462-11ggaqf1 cord-009877-3cyz6o9c cord-009784-kxa68zbc cord-011840-neowfhwg cord-281081-rifr5uub cord-013348-lsksys56 cord-255390-dvp0luxe cord-010578-uib9h1lb cord-277735-a9gkath5 cord-006828-i88on326 cord-015683-a9a82of4 cord-262762-mgegswvn cord-022310-yc6xtw0s cord-022353-q2k2krnm 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cord-344871-486sk4wc cord-344309-6c2wttxg cord-348660-qnbgywgy cord-349744-8cg5yj20 cord-344581-h7ikjgic cord-340960-abanr641 cord-346320-ysgz6adr cord-353614-z4fpy607 cord-351498-bmq6zcb0 cord-348161-757c51xw cord-353190-7qcoxl81 cord-022650-phsr10jp cord-019347-tj3ye1mx cord-022501-9wnmdvg5 cord-023095-4dannjjm cord-015021-pol2qm74 cord-031907-ilhr3iu5 cord-022888-dnsdg04n cord-015394-uj7fe5y6 cord-022940-atbjwpo5 Creating transaction Updating ent table ===== Reducing parts of speech cord-003208-lwirkob3 cord-000434-ff2zadol cord-004101-0r2g5p1i cord-003471-tr3ageky cord-007480-grndfx7b cord-003315-r1wkx0ml cord-003656-7mzsaz7a cord-009922-t1hoox6e cord-007497-nn5l5rai cord-011212-ovjdzyxv cord-007495-gpz4gkv3 cord-003623-n01rgqyv cord-001446-mpuovmeb cord-003859-k8wfyj9b cord-012891-heqsfzkm cord-007644-7bsixsgd cord-009877-3cyz6o9c cord-009784-kxa68zbc cord-011840-neowfhwg cord-281081-rifr5uub cord-013348-lsksys56 cord-255390-dvp0luxe cord-010578-uib9h1lb cord-277735-a9gkath5 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cord-301313-9595vm0k cord-300701-vkzya7uq cord-284841-flhfagp3 cord-281760-34wuttqw cord-297684-9q3oopaz cord-285700-9q6vwoct cord-293393-kbndie8e cord-301655-6nxhvvm4 cord-313517-5ipj2z86 cord-279406-wwdqh9qs cord-311811-nrodyagi cord-291104-6chpmgry cord-301355-9lswjro2 cord-300908-i80tuhqk cord-316723-srenbxa7 cord-301175-6alsigxk cord-317123-0tdfvlqd cord-311349-145kwny3 cord-310536-u30cufg7 cord-309415-77b5erfj cord-318444-sgm24q1i cord-312456-6lxc2rj2 cord-026559-xx52u01h cord-317057-c2bwky6e cord-313193-q5zeoqlb cord-327890-ocisq7e4 cord-334165-7gfk554m cord-317026-9zgc6xrb cord-333026-9f6ecg30 cord-320559-up1q3k6q cord-334277-g3go3u02 cord-326232-668f53qc cord-317223-juw4xt8q cord-318266-i8x80knq cord-305516-s1lvhknm cord-343185-lbmbp9ca cord-334968-gonx5taq cord-321691-46la29tm cord-334896-3g75spkc cord-335591-r0x8yaqj cord-335121-ro3x3qa3 cord-336899-zngp67tb cord-331277-fjsuo3yy cord-322529-3xn5v54s cord-351952-lhhjax3s cord-346574-u28y1ttw cord-347374-mryazbnq cord-342024-kaku49xd cord-344871-486sk4wc cord-344309-6c2wttxg cord-340960-abanr641 cord-349744-8cg5yj20 cord-344581-h7ikjgic cord-348660-qnbgywgy cord-346320-ysgz6adr cord-353614-z4fpy607 cord-351498-bmq6zcb0 cord-348161-757c51xw cord-022653-qa1uph35 cord-353190-7qcoxl81 cord-015147-h0o0yqv8 cord-006860-a3b8hyyr cord-022650-phsr10jp cord-019347-tj3ye1mx cord-023095-4dannjjm cord-022501-9wnmdvg5 cord-001521-l36f1gp7 cord-015021-pol2qm74 cord-022888-dnsdg04n cord-031907-ilhr3iu5 cord-022940-atbjwpo5 cord-015394-uj7fe5y6 Creating transaction Updating pos table Building ./etc/reader.txt cord-022940-atbjwpo5 cord-022650-phsr10jp cord-022888-dnsdg04n cord-317026-9zgc6xrb cord-257715-pbcr81qm cord-023095-4dannjjm number of items: 143 sum of words: 2,427,034 average size in words: 25,547 average readability score: 50 nouns: cells; patients; results; cell; study; protein; virus; expression; levels; samples; serum; treatment; infection; disease; group; analysis; antibodies; methods; blood; mice; antibody; assay; control; response; time; data; activity; influenza; detection; evs; test; proteins; studies; effect; gene; plasma; role; ml; days; antigen; groups; conclusion; effects; age; method; production; viruses; years; system; dogs verbs: used; showed; increases; compared; induced; determine; including; found; detect; performing; associated; based; followed; suggesting; observed; identify; evaluated; developed; obtained; treat; testing; reported; measured; demonstrated; reduce; investigated; expressed; isolated; indicated; binding; containing; decreased; caused; provide; collected; derived; produce; assess; resulting; confirm; revealed; studied; described; infected; involves; known; analyzed; leading; presenting; mediates adjectives: specific; human; positive; different; clinical; high; significant; anti; higher; negative; non; immune; inflammatory; viral; normal; low; important; first; severe; total; present; similar; lower; healthy; new; respiratory; acute; molecular; several; recombinant; diagnostic; allergic; early; common; dependent; single; major; available; potential; fetal; various; many; small; western; mean; novel; chronic; free; cellular; multiple adverbs: also; significantly; however; well; respectively; therefore; previously; highly; recently; furthermore; even; prior; moreover; especially; currently; still; approximately; statistically; often; less; clinically; together; directly; finally; alone; later; specifically; first; mainly; commonly; frequently; particularly; interestingly; additionally; subsequently; now; widely; least; relatively; usually; yet; strongly; potentially; commercially; rapidly; generally; successfully; similarly; almost; naturally pronouns: we; it; our; their; its; they; i; them; he; his; she; her; us; itself; one; themselves; you; your; him; me; https://doi.org/10.1101/2020.06.02.20120345; mg; my; apod; mrnas; iga+; wb103; s; interleukin-15; il-; igfbp2; himself; y€; thier; siil-33; ocid1001; n40np; mrs; itsn2; imagej; igmcic; igg4; esat-6; e2f2-/-mice; crx-527; ykl-40; wi~; tssc; trl2x4; tnf~ proper nouns: ELISA; ⁄; SARS; PCR; mg; T; C; sera; University; RNA; TNF; CoV-2; M; A; IgG; S; LPS; IgE; IFN; RT; S.; USA; H1N1; L; IL-6; E.; B; N; II; EV; Fig; CD4; PBS; CoV; kg; PEDV; AE; IgM; Summary; HA; C.; D; CD8; mRNA; I; H5N1; IgA; M.; G; NA keywords: elisa; sars; pcr; study; cell; result; patient; dna; antibody; protein; pedv; level; university; tnf; method; il-6; western; rna; mouse; ifn; group; expression; disease; conclusion; virus; treatment; pbs; increase; high; cd4; usa; test; response; rbd; mers; lps; introduction; institute; infection; ibv; ebola; blood; year; vaccine; tgev; sample; research; objective; isolate; il-1 one topic; one dimension: cells file(s): https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3158760/ titles(s): Identification of a Highly Conserved H1 Subtype-Specific Epitope with Diagnostic Potential in the Hemagglutinin Protein of Influenza A Virus three topics; one dimension: patients; cells; virus file(s): https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159469/, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480431/, https://doi.org/10.1186/s12917-015-0500-z titles(s): Abstracts TPS | ISEV2020 Abstract Book | Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus five topics; three dimensions: virus elisa influenza; cells evs cell; patients results cells; patients cells cell; il patients mice file(s): https://www.ncbi.nlm.nih.gov/pubmed/18501276/, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480431/, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7104449/, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159469/, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7095072/ titles(s): Diagnostic Investigation of Emerging Viruses of Companion Animals | ISEV2020 Abstract Book | Scientific Abstracts | Abstracts TPS | Third International Congress on the Immune Consequences of Trauma, Shock and Sepsis —Mechanisms and Therapeutic Approaches Type: cord title: keyword-elisa-cord date: 2021-05-24 time: 23:40 username: emorgan patron: Eric Morgan email: emorgan@nd.edu input: keywords:elisa ==== make-pages.sh htm files ==== make-pages.sh complex files ==== make-pages.sh named enities ==== making bibliographics id: cord-280939-d478p8u6 author: Abe, Kento T. title: A simple protein-based surrogate neutralization assay for SARS-CoV-2 date: 2020-10-02 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Most of the patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mount a humoral immune response to the virus within a few weeks of infection, but the duration of this response and how it correlates with clinical outcomes has not been completely characterized. Of particular importance is the identification of immune correlates of infection that would support public health decision-making on treatment approaches, vaccination strategies, and convalescent plasma therapy. While ELISA-based assays to detect and quantitate antibodies to SARS-CoV-2 in patient samples have been developed, the detection of neutralizing antibodies typically requires more demanding cell-based viral assays. Here, we present a safe and efficient protein-based assay for the detection of serum and plasma antibodies that block the interaction of the SARS-CoV-2 spike protein receptor binding domain (RBD) with its receptor, angiotensin-converting enzyme 2 (ACE2). The assay serves as a surrogate neutralization assay and is performed on the same platform and in parallel with an ELISA for the detection of antibodies against the RBD, enabling a direct comparison. The results obtained with our assay correlate with those of 2 viral-based assays, a plaque reduction neutralization test (PRNT) that uses live SARS-CoV-2 virus and a spike pseudotyped viral vector–based assay. url: https://www.ncbi.nlm.nih.gov/pubmed/32870820/ doi: 10.1172/jci.insight.142362 id: cord-346320-ysgz6adr author: Arabi, Yaseen M. title: Feasibility of Using Convalescent Plasma Immunotherapy for MERS-CoV Infection, Saudi Arabia date: 2016-09-17 words: 4239.0 sentences: 203.0 pages: flesch: 49.0 cache: ./cache/cord-346320-ysgz6adr.txt txt: ./txt/cord-346320-ysgz6adr.txt summary: We explored the feasibility of collecting convalescent plasma for passive immunotherapy of Middle East respiratory syndrome coronavirus (MERS-CoV) infection by using ELISA to screen serum samples from 443 potential plasma donors: 196 patients with suspected or laboratory-confirmed MERS-CoV infection, 230 healthcare workers, and 17 household contacts exposed to MERS-CoV. We explored the feasibility of collecting convalescent plasma for passive immunotherapy of Middle East respiratory syndrome coronavirus (MERS-CoV) infection by using ELI-SA to screen serum samples from 443 potential plasma donors: 196 patients with suspected or laboratory-confirmed MERS-CoV infection, 230 healthcare workers, and 17 household contacts exposed to MERS-CoV. In collaboration with the King Abdullah International Medical Research Center, the Gulf Cooperation Council Infection Control Center, and the World Health Organization (WHO)-International Severe Acute Respiratory and Emerging Infection Consortium MERS-CoV Working Group, we developed a study protocol to screen potential donors, collect high-titer convalescent plasma, and administer the plasma in a clinical trial (13) . abstract: We explored the feasibility of collecting convalescent plasma for passive immunotherapy of Middle East respiratory syndrome coronavirus (MERS-CoV) infection by using ELISA to screen serum samples from 443 potential plasma donors: 196 patients with suspected or laboratory-confirmed MERS-CoV infection, 230 healthcare workers, and 17 household contacts exposed to MERS-CoV. ELISA-reactive samples were further tested by indirect fluorescent antibody and microneutralization assays. Of the 443 tested samples, 12 (2.7%) had a reactive ELISA result, and 9 of the 12 had reactive indirect fluorescent antibody and microneutralization assay titers. Undertaking clinical trials of convalescent plasma for passive immunotherapy of MERS-CoV infection may be feasible, but such trials would be challenging because of the small pool of potential donors with sufficiently high antibody titers. Alternative strategies to identify convalescent plasma donors with adequate antibody titers should be explored, including the sampling of serum from patients with more severe disease and sampling at earlier points during illness. url: https://doi.org/10.3201/eid2209.151164 doi: 10.3201/eid2209.151164 id: cord-024171-x7y1xpsf author: Bachmann, P. A. title: Rotavirusnachweis in Faezes: Erfahrungen mit dem Enzyme Linked Immunosorbent Assay (ELISA) date: 2010-05-13 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: ZUSAMMENFASSUNG: 313 Kälber‐, 47 Ferkel‐ und 100 menschliche Faezesproben von an Durchfall erkrankten neugeborenen Patienten wurden mit Hilfe einer modifizierten Immunelektronenmikroskopie‐Technik (IEM) und im Enzyme‐linked immunosorbent assay (ELISA) vergleichend untersucht. In 112 Kälber‐ (36%), 2 Ferkel‐ (4 %) und 21 (21 %) menschlichen Faezesproben konnte Rotavirus oder ‐antigen nachgewiesen werden. Die Sicherheit der Methodik wurde zusätzlich anhand der Virusausscheidung von drei experimentell infizierten, kolostrumfrei aufgezogenen Kälbern überprüft. Der ELISA, bei dem Meerrettichperoxidase als Enzym verwendet wurde, war spezifisch und erwies sich im Vergleich zur IEM als empfindlicher. Bei 18 von insgesamt 136 positiven Proben gelang der Virusnachweis nur mit der IEM, diese Proben waren negativ im ELISA. In diesen Proben werden Virus‐Antikörperkomplexe vermutet. Weitere Untersuchungen sind notwendig, bevor der ELISA zum Rotavirusnachweis in der Routinediagnostik empfohlen werden kann. SUMMARY: Demonstration of Rotavirus in Faeces: Experiences with the Enzyme linked immunosorbent assay (ELISA) 313 calf, 47 piglet and 100 human faecal samples from newborn patients with diarrhea were investigated comparing a modified immune electron microscopy method and the enzyme linked immunosorbent assay (ELISA). In 112 (36 %) calf, 2 (4 %) piglet and 21 (21 %) human samples rotavirus or its antigen could be demonstrated. The suitability of the method was also tested by studying virus shedding of three experimentally infected, colostrum‐deprived calves. The ELISA, using horse‐radish peroxidase as enzyme and 5 amino‐salicylic acid as indicator, was very specific and showed a higher sensitivity compared to IEM. In 18 out of the 136 positive samples rotavirus could only be demonstrated by IEM; these samples were negative in the ELISA. It is assumed that these samples contain virus‐antibody complexes. Further investigations are necessary before the ELISA can be recommended as a routine diagnostic test. RÉSUMÉ: Mise en évidence de Rotavirus dans des matières fécales: Expériences avec ELISA (Enzyme Linked Immunosorbent Assay) 313 maitères fécales de veaux, 47 de porcelets et 100 échantillons humains de patients nouveaux‐nés atteints de diarrhée ont été examinés en comparaison avec une technique immunologique modifiée en microscopie électronique (IEM) et avec ELISA. Rotavirus ou l'antigène ont pu être mis en évidence chez 113 veaux (36%), 2 porcelets (4%) et 21 enfants La fiabilité de la méthode a été en plus testée sur la base de l'excrétion du virus chez trois veaux infectés expérimentalement et privés de colostrum. (21 %). Le test ELISA, l'enzyme utilisée étant la peroxydase de Meerrettich, fut spécifique et s'est révélé plus sensible que IEM. La mise en évidence du virus a été possible dans 18 cas sur 136 positifs avec la méthode IEM, ces échantillons s'étant révélés négatifs avec ELISA. On a supposé l'existence de complexes virus‐anticorps dans ces échantillons. D'autres recherches sont nécessaires avant de pouvoir recommander le test ELISA pour la mise en évidence de Rotavirus dans le diagnostic de routine. RESUMEN: Identificación de virus Rota en heces: Experiencia con el Enzyme Linked Immunosorbent Assay (ELISA) Se examinaron comparativamente 313 muestras de heces de terneros, 47 de lechones y 100 de niños, pacientes recién enfermos de diarrea, con ayuda de una técnica modificada de inmunomicroscopía de electrones (IEM) y en el Enzyme Linked Immunosorbent Assay (ELISA). En 113 muestras de heces de terneros (36 %), 2 de lechones (4 %) y 21 de niños (21 %) se pudo identificar virus o antígeno Rota. La seguridad de la metodología se controló además con ayuda de la eliminación de virus por tres terneros infectados experimentalmente, criados sin calostro. El ELISA, en el que se utilizó peroxidasa de rábano rusticano como enzima, era específico, resultando ser más sensible en comparación con la IEM. Solo en 18 de un total de 136 muestras positivas se logró la identificación de virus con la IEM, siendo estas pruebas negativas en el ELISA. Se sospecha que en estas muestras había complejos de anticuerpos de virus. Se precisa proseguir con las experiencias antes de poder recomendar el ELISA para la identificación de virus Rota en el diagnóstico de rutina. An dieser Stelle möchte ich Dr. D. Ellens und Dr. P. de Leeuw, Central Diergeneeskundig Instituut, Leystad, Holland, sehr herzlich für die Überlassung von Referenzmaterial und die Hilfe bei der Entwicklung des ELISA in unserem Labor danken. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192341/ doi: 10.1111/j.1439-0450.1979.tb00795.x id: cord-009877-3cyz6o9c author: Barclay, Wendy S. title: Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions date: 2005-12-07 words: 3078.0 sentences: 144.0 pages: flesch: 47.0 cache: ./cache/cord-009877-3cyz6o9c.txt txt: ./txt/cord-009877-3cyz6o9c.txt summary: In a preliminary study involving 12 volunteers inoculated intranasally with human rhinovirus EL (HRV-EL), we showed that the presence of circulating rhinovirus-specific IgA, rather than IgG, protected volunteers from infection [Barclay and Al-Nakib, 19871. Figure 1 shows a) the mean HRV-2-neutralising antibody, b) specific IgG and c) IgA in the serum, and d) specific IgA in nasal secretions (normalized for total IgA) in pre-inoculation samples from the three groups of volunteers. Recently, a similar study involving samples obtained from volunteers inoculated with another rhinovirus serotype (HRV-9) and tested by ELISA for homologous antibodies, appears to confirm the association between ELISA-serumspecific IgA and protection against reinfection (Callow and Sergeant, unpublished data). The size of rises in HRV-2-specific IgA in the serum after virus inoculation showed a significant negative correlation with the concentration of specific IgA in the pre-inoculation nasal secretions and with the presence of serum neutralising antibody, indicating that volunteers who were most susceptible to infection and illness, in fact, showed the largest antibody response. abstract: Rhinovirus‐specific antibodies have traditionally been detected by their ability to neutralise the homologous rhinovirus serotype in tissue culture. Recently, however, we have described an enzyme‐linked immunosorbent assay that detects rhinovirus‐specific antibodies in sera and nasal secretions [Barclay and Al‐Nakib, 1987]. Here we describe an evaluation of the ELISA in a study involving 71 adult volunteers inoculated intranasally with human rhinovirus type 2 (HRV‐2). Pre‐and post‐inoculation serum samples and pre‐inoculation nasal washings were tested for the presence of HRV‐2‐specific antibodies by ELISA. Such antibodies were associated with protection against infection when present locally in nasal secretions, but when also present in the serum they were associated with protection against both infection and the development of illness. The antibody concentrations showed strong correlation with each other and with that of antibodies detected by the neutralisation test. Following HRV‐2 infection, rises in HRV‐2‐specific IgA in sera detected by ELISA occurred more frequently than rises in neutralizing antibody. These results suggest that the ELISA is a sensitive and reliable indicator of recent infection, as well as a predictor of homologous immune status. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7166658/ doi: 10.1002/jmv.1890250411 id: cord-301974-4wn40ivq author: Berry, Jody D title: Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus date: 2004-09-01 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development. url: https://www.ncbi.nlm.nih.gov/pubmed/15234813/ doi: 10.1016/j.jviromet.2004.04.009 id: cord-012891-heqsfzkm author: Blanco Vázquez, Cristina title: Detection of latent forms of Mycobacterium avium subsp. paratuberculosis infection using host biomarker-based ELISAs greatly improves paratuberculosis diagnostic sensitivity date: 2020-09-03 words: 7818.0 sentences: 354.0 pages: flesch: 43.0 cache: ./cache/cord-012891-heqsfzkm.txt txt: ./txt/cord-012891-heqsfzkm.txt summary: The aim of this study was to evaluate the diagnostic potential of commercial ELISAs based on detection of these five bovine biomarkers to detect latent and patent forms of MAP infection in naturally infected cattle, using reference serum samples from well characterized animals with focal, multifocal and diffuse histological lesions in their intestinal or associated lymphoid tissues [40]. The diagnostic performance of the ABCA13, MMP8 and SPARC-based ELISAs for the focal and any type of lesion groups was compared with the IDEXX ELISA and additionally with other conventional PTB diagnosis methods such as specific fecal and tissue real-time PCR and bacteriological culture ( Table 4 ). Our results indicate that the ABCA13, SPARC and MMP8-based ELISAs have higher AUC values and sensitivities than the IDEXX ELISA and other current diagnostic methods for detection of animals with focal, multifocal and any type of histopathological lesions, respectively. abstract: Bovine paratuberculosis (PTB) is a chronic granulomatous enteritis, caused by Mycobacterium avium subsp. paratuberculosis (MAP), responsible for important economic losses in the dairy industry. Current diagnostic methods have low sensitivities for detection of latent forms of MAP infection, defined by focal granulomatous lesions and scarce humoral response or MAP presence. In contrast, patent infections correspond to multifocal and diffuse types of enteritis where there is increased antibody production, and substantial mycobacterial load. Our previous RNA-Seq analysis allowed the selection of five candidate biomarkers overexpressed in peripheral blood of MAP infected Holstein cows with focal (ABCA13 and MMP8) and diffuse (FAM84A, SPARC and DES) lesions vs. control animals with no detectable PTB-associated lesions in intestine and regional lymph nodes. The aim of the current study was to assess the PTB diagnostic potential of commercial ELISAs designed for the specific detection of these biomarkers. The ability of these ELISAs to identify animals with latent and/or patent forms of MAP infection was investigated using serum from naturally infected cattle (n = 88) and non-infected control animals (n = 67). ROC analysis revealed that the ABCA13-based ELISA showed the highest diagnostic accuracy for the detection of infected animals with focal lesions (AUC 0.837, sensitivity 79.25% and specificity 88.06%) and with any type of histological lesion (AUC 0.793, sensitivity 69.41% and specificity 86.57%) improving on the diagnostic performance of the popular IDEXX ELISA and other conventional diagnostic methods. SPARC and MMP8 showed the highest diagnostic accuracy for the detection of animals with multifocal (AUC 0.852) and diffuse lesions (AUC 0.831), respectively. In conclusion, our results suggest that quantification of ABCA13, SPARC and MMP8 by ELISA has the potential for implementation as a diagnostic tool to reliably identify MAP infection, greatly improving early detection of MAP latent infections when antibody responses and fecal shedding are undetectable using conventional diagnostic methods. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7470414/ doi: 10.1371/journal.pone.0236336 id: cord-009784-kxa68zbc author: Bolton, Jessica S. title: Comparison of ELISA with electro-chemiluminescence technology for the qualitative and quantitative assessment of serological responses to vaccination date: 2020-04-17 words: 6166.0 sentences: 286.0 pages: flesch: 47.0 cache: ./cache/cord-009784-kxa68zbc.txt txt: ./txt/cord-009784-kxa68zbc.txt summary: The PfCSP-FL protein is comprised of 26 Tyr -127 Asp linked to 207 Pro -383 Ser [4] ; "Repeat" is a Keywords: Serology, Vaccine, Antigen, Multiplex, Antigenic competition, ELISA, Electro-chemiluminescence 32-mer peptide representing the central Repeat region (NANP 8 ); C-term is a recombinant protein representing the C-terminal fragment (AA 207-383); Pf16 is an epitope within the C-terminus that has been used as a functional marker when evaluating anti-CSP antibodies induced by vaccination [4, 7, 8] . To establish a multiplex assay using an ECLIA platform, several parameters (i.e., antigen coating concentration, antigenic competition between closely related antigens, sample dilutions) were optimized and the performance of the assay determined in regards to specificity, linearity, and throughput. abstract: BACKGROUND: Profiling immune responses induced by either infection or vaccination can provide insight into identification of correlates of protection. Furthermore, profiling of serological responses can be used to identify biomarkers indicative of exposure to pathogens. Conducting such immune surveillance requires readout methods that are high-throughput, robust, and require small sample volumes. While the enzyme-linked immunosorbent assay (ELISA) is the classical readout method for assessing serological responses, the advent of multiplex assays has significantly increased the throughput and capacity for immunoprofiling. This report describes the development and assay performance (sensitivity, linearity of detection, requirement for multiple dilutions for each sample, intra- and inter-assay variability) of an electro-chemiluminescence (ECLIA)-based multiplex assay. METHODS: The current study describes the development of a multiplex ECLIA-based assay and characterizes the sensitivity, linear range, and inter- and intra-assay variability of the ECLIA platform and its agreement with the traditional ELISA. Special emphasis was placed on potential antigenic competition when testing closely related antigens in the multiplex format. RESULTS: Multiplexing of antigens in ECLIA provides significant practical benefits in terms of reducing sample volume requirements and experimental time. Beyond the practical advantages of multiplexing, the ECLIA provides superior assay performance when compared to the ELISA. Not only does ECLIA show good agreement with the ELISA assay, but the linear range of ECLIA is also sufficiently wide to permit single-dilution measurements of concentration without the need to do serial dilutions. The lack of antigenic competition allows the simultaneous testing of closely related antigens, such as plate antigens representing different alleles of the same protein, which can inform about cross-reactivities—or lack thereof—of serological responses. CONCLUSION: The advantages of the newly developed tool for assessing the antigen profiles of serological responses may ultimately lead to the identification of biomarkers associated with various disease stages and or protection against disease. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7165447/ doi: 10.1186/s12936-020-03225-5 id: cord-293393-kbndie8e author: Braesch-Andersen, Sten title: ApoD Mediates Binding of HDL to LDL and to Growing T24 Carcinoma date: 2014-12-16 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Apolipoprotein (Apo) D is an important protein produced in many parts of the body. It is necessary for the development and repair of the brain and protection from oxidative stress. The purpose of this study was to investigate the extent to which apoD interacts with lipoproteins in human plasma. By using detergent-free ELISA, we show that immobilized monoclonal antibodies against apoD very efficiently bind to low density lipoprotein (LDL) from plasma; this binding is as equally efficient as binding to an anti-apoB monoclonal antibody. Adding detergent to the plasma inhibited the binding, suggesting that the binding is dependent on the presence of intact lipoprotein particles. Reversing the system by using immobilized anti-apoB revealed that the affinity of apoD for LDL is rather low, suggesting that multiple bindings are needed for a durable connection. Biosensor experiments using purified lipoproteins also showed that purified apoD and high density lipoprotein 3 (HDL3), a lipoprotein fraction rich in apoD, were both able to bind LDL very efficiently, indicating that the HDL3-LDL interaction may be a physiological consequence of the affinity of apoD for LDL. Furthermore, we found that apoD increases the binding of HDL to actively growing T24 bladder carcinoma cells but not to quiescent, contact-inhibited, confluent T24 cells. This result is especially intriguing given that the T24 supernatant only contained detectable levels of apoD after growth inhibition, raising the possibility that alternating the expression of apoD and a putative apoD-receptor could give direction to the flow of lipids. In the current paper, we conclude that apoD mediates binding of HDL to LDL and to growing T24 carcinomas, thereby highlighting the importance of apoD in lipid metabolism. url: https://doi.org/10.1371/journal.pone.0115180 doi: 10.1371/journal.pone.0115180 id: cord-001446-mpuovmeb author: Bratcher, Preston E. title: Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA date: 2014-11-03 words: 4873.0 sentences: 226.0 pages: flesch: 43.0 cache: ./cache/cord-001446-mpuovmeb.txt txt: ./txt/cord-001446-mpuovmeb.txt summary: Circulating levels of SP-D have been examined for their potential use as a biomarker in various diseases including dermatitis [2, 3] , acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] , periodontitis [14] , interstitial pulmonary fibrosis (IPF) [10, 12, [15] [16] [17] [18] [19] [20] [21] [22] [23] , chronic obstructive pulmonary disease (COPD) [15, [24] [25] [26] [27] [28] [29] [30] [31] [32] [33] [34] [35] [36] , emphysema [37] , cystic fibrosis (CF) [15, 38, 39] , coronary disease [40, 41] , sclerosis [42] [43] [44] [45] [46] , cancer [47, 48] , sarcoidosis [21, 49] , allergies [28, [50] [51] [52] , rheumatoid arthritis [53, 54] , and respiratory infections [18, [55] [56] [57] [58] [59] [60] . Serum levels of surfactant proteins A and D are useful biomarkers for interstitial lung disease in patients with progressive systemic sclerosis abstract: BACKGROUND: Extensive variations in human surfactant protein D (SP-D) levels in circulation as measured by ELISA exist in the published literature. In order to determine the source of these variations, factors influencing the measurement by ELISA were explored. MATERIALS AND METHODS: Peripheral blood from healthy individuals was collected into various vacutainers during the same blood draw. Recombinant SP-D was diluted into different matrices and used for a standard curve. Samples were analyzed by capture ELISA using one of two distinct detection antibodies. RESULTS: The type of matrix had some effects on detection of recombinant SP-D. The type of anticoagulant used and dilution factor had very little effect, except for in plasma collected in EDTA vacutainers. The extent of variation in published values seemed to be due to the ELISA configuration employed, and, in agreement with this, we found that by switching the detection antibody, there was a 50% decrease in the extrapolated SP-D value of serum and plasma samples. Storage of samples resulted in slight changes in measured SP-D levels. CONCLUSIONS: The ELISA configuration employed to measure circulating levels of SP-D has a significant effect on the extrapolated values. In both configurations tested, the use of EDTA as a coagulant resulted in inconsistent values, and we, therefore, suggest the avoidance of this anticoagulant when assaying for SP-D by ELISA. While the demonstrated effects of several factors on measurement of SP-D may not account for all the disparities amongst the previous studies, they stress that variations in methodologies for measuring the same protein can result in very inconsistent results. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4218753/ doi: 10.1371/journal.pone.0111466 id: cord-340960-abanr641 author: Brigger, D. title: Accuracy of serological testing for SARS‐CoV‐2 antibodies: first results of a large mixed‐method evaluation study date: 2020-09-30 words: 4479.0 sentences: 289.0 pages: flesch: 50.0 cache: ./cache/cord-340960-abanr641.txt txt: ./txt/cord-340960-abanr641.txt summary: In a mixed‐design evaluation study, we compared the diagnostic accuracy of serological immunoassays that are based on various SARS‐CoV‐2 proteins and assessed the neutralizing activity of antibodies in patient sera. A total of 54 randomly selected sera from individuals who were tested positive in either of the three ELISA immunoassays as well as 6 negative controls were assessed in a live SARS-CoV-2 neutralization assay (all collected in April 2020). Recombinantly expressed RBD has been used to establish an in-house ELISA for the detection of IgM and IgG anti-SARS-CoV-2 antibodies in human serum samples (supplementary Fig. 1a,b) . A total of 54 randomly selected sera from individuals who were tested positive in either of the three ELISA immunoassays as well as 6 negative controls were assessed in a live SARS-CoV-2 neutralization assay using ACE2-expressing Vero-E6 cells (34 inpatient samples, and 26 samples of medical personnel). abstract: BACKGROUND: Serological immunoassays that can identify protective immunity against SARS‐CoV‐2 are needed to adapt quarantine measures, assess vaccination responses, and evaluate donor plasma. To date, however, the utility of such immunoassays remains unclear. In a mixed‐design evaluation study, we compared the diagnostic accuracy of serological immunoassays that are based on various SARS‐CoV‐2 proteins and assessed the neutralizing activity of antibodies in patient sera. METHODS: Consecutive patients admitted with confirmed SARS‐CoV‐2 infection were prospectively followed alongside medical staff and biobank samples from winter 2018/2019. An in‐house enzyme‐linked immunosorbent assay utilizing recombinant receptor‐binding domain (RBD) of the SARS‐CoV‐2 spike protein was developed and compared to three commercially available enzyme‐linked immunosorbent assays (ELISAs) targeting the nucleoprotein (N), the S1 domain of the spike protein (S1) and a lateral flow immunoassay (LFI) based on full‐length spike protein. Neutralization assays with live SARS‐CoV‐2 were performed. RESULTS: One‐thousand four‐hundred and seventy‐seven individuals were included comprising 112 SARS‐CoV‐2 positives (defined as a positive real‐time PCR result; prevalence 7.6%). IgG seroconversion occurred between day 0 and day 21. While the ELISAs showed sensitivities of 88.4% for RBD, 89.3% for S1, and 72.9% for N protein, the specificity was above 94% for all tests. Out of 54 SARS‐CoV‐2 positive individuals, 96.3% showed full neutralization of live SARS‐CoV‐2 at serum dilutions ≥1:16, while none of the 6 SARS‐CoV‐2 negative sera revealed neutralizing activity. CONCLUSIONS: ELISAs targeting RBD and S1 protein of SARS‐CoV‐2 are promising immunoassays which shall be further evaluated in studies verifying diagnostic accuracy and protective immunity against SARS‐CoV‐2. url: https://doi.org/10.1111/all.14608 doi: 10.1111/all.14608 id: cord-318266-i8x80knq author: Böse, Reinhard title: Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot date: 1994-05-31 words: 3622.0 sentences: 220.0 pages: flesch: 61.0 cache: ./cache/cord-318266-i8x80knq.txt txt: ./txt/cord-318266-i8x80knq.txt summary: title: Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. Sera tested were as follows: (I) sera from 106 horses not infected with Bubesia spp., i.e. horses from Germany tested negative in preliminary studies by IFAT at a serum dilution of 4 l/40 and by CFT at a serum dilution of < l/5; the results for these sera were used to calculate the diagnostic specificity of the ELISA and Western blot; (2) 71 sera from 15 horses experimentally infected with B. equi; these results were used to calculate the diagnostic sensitivity of the ELISA, Western blot, IFAT and CFT; (3) 76 sera from 13 horses experimentally infected with 3. abstract: Abstract From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofluorescence antibody test (IFAT) and complement fixation test (CFT). The sensitivity of the ELISA of 98.3% obtained for sera from day 14 after infection was superior to the Western blot (94.9%), the IFAT (96.6%) and the CFT (28.8%). No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. resulting in a calculated specificity of 100% for both tests. Cross reactions of B. equi-positive sera did occur to a larger extent in the ELISA (20%) than in the IFAT (4%). No cross reactions were observed with the Western blot and the CFT. The higher sensitivity of the ELISA was also demonstrated by testing of 132 field sera: more positive results were obtained by ELISA (112) as compared to IFAT (92) or CFT (41). The validity of these results was confirmed by testing of sera by Western blot. The ELISA as the most sensitive test provides the best method for the identification of carrier horses to prevent the introduction into non-endemic areas (export testing). Positive ELISA results can be confirmed by Western blot, if a species-specific diagnosis is required. url: https://www.sciencedirect.com/science/article/pii/0020751994900817 doi: 10.1016/0020-7519(94)90081-7 id: cord-262762-mgegswvn author: Callebaut, P. title: Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea date: 1982-09-30 words: 3605.0 sentences: 179.0 pages: flesch: 50.0 cache: ./cache/cord-262762-mgegswvn.txt txt: ./txt/cord-262762-mgegswvn.txt summary: Abstract An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the coronavirus-like agent in feces of pigs naturally affected with porcine epidemic diarrhea (PED) or experimentally infected with the CV777 isolate. Control fecal samples, used to determine the limit between positive and negative absorbance values in ELISA, consisted of 25 non-diarrheal specimens obtained from conventional pigs of all ages. A total of 62 serum samples, to be examined for antibody by ELISA blocking, were collected from four CDCD piglets and seven conventional experimental fattening pigs prior to inoculation with CV777 and at various intervals between 7 and 81 days thereafter. To compare the sensitivity of EM and ELISA for the detection of CVLA, the specimens of fecal material and intestinal contents obtained from experimentally infected CDCD piglets and experimental fattening pigs as described in the section "Specimens from experimental pigs", were examined for coronavirus by EM. abstract: Abstract An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the coronavirus-like agent in feces of pigs naturally affected with porcine epidemic diarrhea (PED) or experimentally infected with the CV777 isolate. The assay was specific and more sensitive than electron microscopy. An ELISA blocking assay is described for the detection and titration of antibodies. Specific antibody formation was demonstrated in pigs experimentally infected with CV777 and in swine naturally affected with PED. url: https://www.sciencedirect.com/science/article/pii/0378113582900098 doi: 10.1016/0378-1135(82)90009-8 id: cord-281393-96j70n2z author: Capai, L. title: Seroprevalence of SARS-CoV-2 IgG antibodies, in Corsica (France), April and June 2020. date: 2020-09-30 words: 3684.0 sentences: 262.0 pages: flesch: 60.0 cache: ./cache/cord-281393-96j70n2z.txt txt: ./txt/cord-281393-96j70n2z.txt summary: A minimum sample size of 1814 was calculated assuming an a priori 5% IgG anti-SARS-CoV-2 seroprevalence (Salje et al., 2020) , a confidence in the estimate of 95%, a maximum allowable error in the prevalence of 1%, and a Corsican population size of 344,679 habitants based on the latest French census data (INSEE, 2020). Residual sera obtained from persons of all ages were tested for the presence of anti-SARS-CoV-2 IgG using the EUROIMMUN enzyme immunoassay kit for semiquantitative detection of IgG antibodies against S1 domain of viral spike protein (ELISA-S) (reference: In all samples with a ratio ≥ 0.8, neutralizing antibodies were detected using a VNT as previously described (Gallian et al., 2020) . To the best of our knowledge this is the first study describing the prevalence of SARS-CoV-2 antibodies in a representative sample of Corsican patients having carried out a blood analysis in biological laboratories after the COVID19 epidemic period. abstract: Our aim was to assess the seroprevalence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection after the lockdown in a sample of the Corsican population. Between 16th April and 15th June 2020, 2,312 residual sera were collected from patients having carried out a blood analysis in one of the participating laboratories. Residual sera obtained from persons of all ages were tested for the presence of anti-SARS-CoV-2 IgG using the EUROIMMUN enzyme immunoassay kit for semiquantitative detection of IgG antibodies against S1 domain of viral spike protein (ELISA-S). Borderline and positive samples in ELISA-S were also tested with an in-house virus neutralization test (VNT). Prevalence values were adjusted for sex and age. A total of 1,973 residual sera samples were included in the study. The overall seroprevalence based on ELISA-S was 5.27% [95% confidence interval (CI) 4.33-6.35] and 5.46% [4.51-6.57] after adjustment. Gender was not associated with IgG detection. However, significant differences were observed between age groups (p-value = 1 E-5) and particularly for people being younger than 50 years of age (Odd ratio (OR) = 2.86 95% CI [1.80- 4.53]; p-value <0.000001*). The prevalence of neutralizing antibody titers [≥]40 was of 3% [2.28-3.84]. In conclusion the present study showed that a low seroprevalence for COVID-19 in Corsica in accordance with values reported for other French regions in which the impact of the pandemic was low. url: https://doi.org/10.1101/2020.09.29.20201368 doi: 10.1101/2020.09.29.20201368 id: cord-313193-q5zeoqlb author: Carrat, F. title: Seroprevalence of SARS-CoV-2 among adults in three regions of France following the lockdown and associated risk factors: a multicohort study. date: 2020-09-18 words: 4076.0 sentences: 265.0 pages: flesch: 57.0 cache: ./cache/cord-313193-q5zeoqlb.txt txt: ./txt/cord-313193-q5zeoqlb.txt summary: Methods Participants in a survey on COVID-19 from an existing consortium of three general adult population cohorts living in the Ile-de-France (IDF) or Grand Est (GE), two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA), with a low rate, were asked to take a dried-blood spot (DBS) for anti-SARS-CoV-2 antibodies assessment. Participants in a survey on COVID-19 from an existing consortium of three general adult population cohorts living in the Ile-de-France (IDF) or Grand Est (GE) -two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA) -with a low rate, were asked to take a dried-blood spot (DBS) for anti-SARS-CoV-2 antibodies assessment. Participants with a positive ELISA-S had a higher rate of self-reported symptoms than those with negative tests except for skin lesions (table 2) is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint abstract: Aim To estimate the seroprevalence of SARS-CoV-2 infection in May-June 2020 after the lockdown in adults living in three regions in France and to identify the associated risk factors. Methods Participants in a survey on COVID-19 from an existing consortium of three general adult population cohorts living in the Ile-de-France (IDF) or Grand Est (GE), two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA), with a low rate, were asked to take a dried-blood spot (DBS) for anti-SARS-CoV-2 antibodies assessment. The primary outcome was a positive anti-SARS-CoV-2 ELISA IgG result against the spike protein of the virus (ELISA-S). The secondary outcomes were a positive ELISA IgG against the nucleocapsid protein (ELISA-NP), anti-SARS-CoV-2 neutralizing antibodies titers >=40 (SN), and predicted positivity obtained from a multiple imputation model (MI). Prevalence estimates were adjusted using sampling weights and post-stratification methods. Findings Between May 4, 2020 and June 23, 2020, 16,000 participants were asked to provide DBS, and 14,628 were included in the analysis, 983 with a positive ELISA-S, 511 with a positive ELISA-NP, 424 with SN>=40 and 941 (Standard Deviation=31) with a positive MI. Adjusted estimates of seroprevalence (positive ELISA-S) were 10.0% (95%CI 9.1%;10.9%) in IDF, 9.0% (95%CI 7.7%; 10.2%) in GE and 3.1% (95%CI 2.4%; 3.7%), in NA. The adjusted prevalence of positive ELISA-NP, SN and MI were 5.7%, 5.0% and 10.0% in IDF, 6.0%, 4.3% and 8.6% in GE, and 0.6%, 1.3% and 2.5% in NA, respectively. A higher seroprevalence was observed in younger participants and when at least one child or adolescent lived in the same household. A lower seroprevalence was observed in smokers compared to non-smokers. Interpretation At the end of the lockdown the prevalence of anti-SARS-CoV-2 IgG or neutralizing antibodies remained low in the French adult population, even in regions with high reported rates of COVID-19. url: https://doi.org/10.1101/2020.09.16.20195693 doi: 10.1101/2020.09.16.20195693 id: cord-346574-u28y1ttw author: Chen, Keyan title: Development and evaluation of an immunochromatographic strip for rapid detection of porcine hemagglutinating encephalomyelitis virus date: 2012-08-24 words: 5509.0 sentences: 277.0 pages: flesch: 50.0 cache: ./cache/cord-346574-u28y1ttw.txt txt: ./txt/cord-346574-u28y1ttw.txt summary: At present, various laboratory methods are available for the detection and surveillance of PHE-CoV, including virus isolation [1] , hemagglutination/hemagglutination inhibition (HA/HI) tests [13] , immunohistochemistry (IHC) assays [10] , and molecular tools such as nestedpolymerase chain reaction (nested PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR) that enable detection of specific CoV RNA sequences from infected tissues [14, 15] . (1) (2) (3) In this study, an immunochromatographic strip with high sensitivity and specificity was developed for the detection of PHE-CoV, combining monoclonal antibody (MAb) and colloidal gold immunochromatography (GICA), and the resulting product is suitable for the surveillance of PHE-CoV. Thus, a lot of brain tissue samples were collected from deceased piglets with suspected PHE-CoV infection, and using RT-PCR and ELISA as reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. abstract: BACKGROUND: The incidence of PHE among pigs in many countries is on the rise, and it has caused great economic losses to the pig industry. Therefore, the development of a sensitive, specific, and easily-performed assay is crucial for the rapid detection and surveillance of PHE-CoV infection and transmission. RESULTS: An immunochromatographic strip was developed for the detection of PHE-CoV. The colloidal gold-labeled MAb 4D4 was used as the detection reagent, and the MAb 1E2 and goat anti-mouse IgG coated the strip's test and control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHE-CoV with a HA unit of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity or specificity. Using RT-PCR as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. There was an excellent agreement between the results obtained by RT-PCR and the immunochromatographic strips (kappa = 0.976). Additionally, there was a strong agreement between the sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatographic strips (Kappa = 0.976). When the immunochromatographic strips were used for diagnosing PHE-CoV infection in the Jilin Province, the PHE-CoV-positive rate ranged from 61.54% in the Jilin district to 17.95% in the Songyuan district. CONCLUSIONS: Based on its high specificity, sensitivity, and stability, the immunochromatographic strip would be suitable for on-site detection of PHE-CoV for surveillance and epidemiological purposes. url: https://doi.org/10.1186/1743-422x-9-172 doi: 10.1186/1743-422x-9-172 id: cord-273361-i5v5rz4x author: Chen, Tsu-Han title: Development of a Luminex assay for the detection of swine antibodies to non-structural proteins of foot-and-mouth disease virus date: 2013-10-31 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Abstract Foot-and mouth disease (FMD), swine vesicular disease (SVD), and vesicular stomatitis (VS) are highly contagious vesicular diseases of swine but are not easy to differentiate clinically. For the purpose of instant detecting of FMD and differentiating it from the other vesicular diseases, a Luminex assay was developed. Sera from 64 infected, 307 vaccinated, and 280 naïve pigs were tested by the Luminex assay. Diagnostic sensitivity of the assay was 100%. Diagnostic specificity of the assay was 98.7% in vaccinated pigs and 97.5% to 100% in naïve pigs. Agreement between the results from the Luminex assay and those from a 3ABC polypeptide blocking ELISA was 96.3% with kappa statistics of 0.92. The Luminex assay can detect the immune response to NSP-3ABC in swine as early as eight days post-infection. Moreover, all of the 15 vaccinated but unprotected pigs were all detected by the Luminex assay. The results indicated that the Luminex assay has potential with specificity in detecting antibodies to FMDV 3ABC NSP and in distinguishing FMDV-infected pigs from with either SVDV or VSV. url: https://www.sciencedirect.com/science/article/pii/S0022175913002251 doi: 10.1016/j.jim.2013.08.002 id: cord-007644-7bsixsgd author: Chirnside, E.D. title: Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus date: 2000-04-04 words: 4054.0 sentences: 198.0 pages: flesch: 49.0 cache: ./cache/cord-007644-7bsixsgd.txt txt: ./txt/cord-007644-7bsixsgd.txt summary: authors: Chirnside, E.D.; Francis, P.M.; De Vries, A.A.F.; Sinclaira, R.; Mumford, J.A. title: Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus A recombinant glutathione-S-transferase fusion protein expressing amino acids 55–98 of equine arteritis virus (EAV) G(L) (rG(L)55–98) was tested in an ELISA for its ability to detect serum antibodies to EAV. This paper describes an indirect ELISA using a recombinant glutathione-Stransferase fusion protein (Smith and Johnson, 1988) as an antigen to screen equine sera for the presence of antibodies to EAV, and its evaluation as a diagnostic test with large numbers of equine serum samples. By testing > 1500 equine sera in ELISA to G,55-98, we have demonstrated that amino acid residues 55-98 of the Bucyrus strain of EAV G,_ encompass a highly immunoreactive antigen which correlates closely with the host virus neutralizing response. abstract: A recombinant glutathione-S-transferase fusion protein expressing amino acids 55–98 of equine arteritis virus (EAV) G(L) (rG(L)55–98) was tested in an ELISA for its ability to detect serum antibodies to EAV. Host antibodies induced following EAV infection bound the recombinant antigen by ELISA. The ELISA specificity and sensitivity were determined with a panel of equine sera including postinfection and postvaccination samples. A good correlation existed between EAV neutralizing antibody titers and ELISA absorbance values (r = 0.827). The sensitivity and specificity of the ELISA were 99.6 and 90.1%, respectively, compared with the EAV neutralization test and the recombinant antigen did not crossreact in ELISA with equine sera directed against other common equine respiratory viruses. Three post-EAV infection equine sera raised against different EAV isolates reacted strongly in the ELISA, as did two equine sera raised against EAV vaccines, indicating that the viral epitope was conserved between the viruses tested. Following vaccination with an inactivated whole virus vaccine, antibody detected with the recombinant antigen ELISA preceded the development of a virus-neutralizing response. The study demonstrates the potential application of rG(L)55–98 as a diagnostic antigen. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7119792/ doi: 10.1016/0166-0934(95)00020-u id: cord-009922-t1hoox6e author: Dearden, C. J. title: Direct detection of rhinoviruses by an enzyme‐linked immunosorbent assay date: 2005-12-07 words: 4065.0 sentences: 205.0 pages: flesch: 55.0 cache: ./cache/cord-009922-t1hoox6e.txt txt: ./txt/cord-009922-t1hoox6e.txt summary: This paper describes the first enzyme‐linked immunosorbent assay for the detection of rhinovirus antigens in clinical specimens (nasal washings), either directly or following overnight cell culture amplification. Secondly, a direct ELISA system was developed in which the nasal washings or control antigen (uninfected tissue culture fluid) were added directly to each of a set of duplicate ELISA plate wells coated with either pre-or postchallenge rabbit anti HRV-EL hyperimmune serum. Figure 2 shows the results obtained with nasal washings, collected from three volunteers on consecutive days following HKV-EL or saline challenge, tested in both the cell-culture-amplified (CCA)-ELISA and direct ELISA systems. Although we would like to emphasise that our data are preliminary, both the direct and CCA-ELISAs gave a good correlation with virus isolation when used to detect rhinovirus antigens in nasal washings (obtained from 18 volunteers challenged with either HRV-EL or saline). abstract: This paper describes the first enzyme‐linked immunosorbent assay for the detection of rhinovirus antigens in clinical specimens (nasal washings), either directly or following overnight cell culture amplification. The assay takes approximately 48 hours to perform and utilizes the same rabbit antirhinovirus hyperimmune serum as both the capture and detecting antibody. The latter has been biotin‐labelled and is detected via a streptavidin 3‐galactosidase preformed complex. This new assay has been found to be very sensitive, detecting human rhinovirus (HRV)‐EL and HRV‐2 at titres as low as 10(1.8) TCID(50) 100 μl(−1) and < 10(1) TCID(50) 100 μl(−1), respectively. Furthermore, when 57 different human rhinovirus serotypes were tested in both the HRV‐EL and HRV‐2 ELISA systems a total of 49 (86%) were found to be cross‐reactive. Of 36 clinical specimens tested by virus isolation, cell‐culture‐amplified (CCA) ELISA, and direct ELISA, 15 were positive by isolation, 11 by CCA‐ELISA, and 11 by direct ELISA. The overall correlation of the CCA and direct ELISA techniques with virus isolation was found to be 88.9% and 66.7%, respectively. The present study demonstrates that the ELISA system developed is a sensitive technique for the diagnosis of rhinovirus infections. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7166933/ doi: 10.1002/jmv.1890230211 id: cord-281081-rifr5uub author: Deng, Junhua title: Serological survey of SARS‐CoV‐2 for experimental, domestic, companion and wild animals excludes intermediate hosts of 35 different species of animals date: 2020-05-07 words: 1515.0 sentences: 86.0 pages: flesch: 55.0 cache: ./cache/cord-281081-rifr5uub.txt txt: ./txt/cord-281081-rifr5uub.txt summary: In this study, 1,914 serum samples from 35 animal species were used for detection of SARS‐CoV‐2‐specific antibodies using double‐antigen sandwich ELISA after validating its specificity and sensitivity. The results showed that no SARS‐CoV‐2‐specific antibodies were detected in above samples which excluded the possibility of 35 animal species as intermediate host for SARS‐CoV‐2. The results showed that no SARS-CoV-2-specific antibodies were detected in above species of animals including pangolin which has been reported as an intermediate host of SARS-CoV-2 (Kangpeng Xiao, 2020) . After confirming the specificity, sensitivity and suitability of SARS-CoV-2 ELISA kit for different species of experimental animals, clinical serum samples from domestic livestock (pig, cow, sheep, horse), poultry (chicken, duck, goose), experimental animal (mice, rat and rhesus monkey), companion animal (dog and cat) and wild animals (camel, fox, mink, alpaca, ferret, bamboo rat, peacock, eagle, tiger rhinoceros, pangolin, leopard cat, jackal, giant panda, masked civet, porcupine, bear, yellow-throated marten, weasel, red pandas and wild boar) were used for antibody detection. abstract: The pandemic SARS‐CoV‐2 has been reported in 123 countries with more than 5,000 patients died from it. However, the original and intermediate hosts of the virus remain unknown. In this study, 1,914 serum samples from 35 animal species were used for detection of SARS‐CoV‐2‐specific antibodies using double‐antigen sandwich ELISA after validating its specificity and sensitivity. The results showed that no SARS‐CoV‐2‐specific antibodies were detected in above samples which excluded the possibility of 35 animal species as intermediate host for SARS‐CoV‐2. More importantly, companion animals including pet dogs (including one dog the SARS‐CoV‐2 patient kept and two dogs which had close contact with it) and cats, street dogs and cats also showed serological negative to SARS‐CoV‐2, which relieved the public concerns for the pets as SARS‐CoV‐2 carriers. url: https://doi.org/10.1111/tbed.13577 doi: 10.1111/tbed.13577 id: cord-297684-9q3oopaz author: Dobaño, Carlota title: Highly sensitive and specific multiplex antibody assays to quantify immunoglobulins M, A and G against SARS-CoV-2 antigens date: 2020-06-12 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Reliable serological tests are required to determine the prevalence of antibodies against SARS-CoV-2 antigens and to characterise immunity to the disease in order to address key knowledge gaps in the context of the COVID-19 pandemic. Quantitative suspension array technology (qSAT) assays based on the xMAP Luminex platform overcome the limitations of rapid diagnostic tests and ELISA with their higher precision, dynamic range, throughput, miniaturization, cost-efficacy and multiplexing capacity. We developed three qSAT assays to detect IgM, IgA and IgG to a panel of eight SARS-CoV-2 antigens including spike (S), nucleoprotein (N) and membrane (M) protein constructs. The assays were optimized to minimize processing time and maximize signal to noise ratio. We evaluated the performance of the assays using 128 plasmas obtained before the COVID-19 pandemic (negative controls) and 115 plasmas from individuals with SARS-CoV-2 diagnosis (positive controls), of whom 8 were asymptomatic, 58 had mild symptoms and 49 were hospitalized. Pre-existing IgG antibodies recognizing N, M and S2 proteins were detected in negative controls suggestive of cross-reactive to common cold coronaviruses. The best performing antibody isotype/antigen signatures had specificities of 100% and sensitivities of 94.94% at ≥14 days since the onset of symptoms and 96.08% at ≥21 days since the onset of symptoms, with AUC of 0.992 and 0.999, respectively. Combining multiple antibody markers as assessed by qSAT assays has the highest efficiency, breadth and versatility to accurately detect low-level antibody responses for obtaining reliable data on prevalence of exposure to novel pathogens in a population. Our assays will allow gaining insights into antibody correlates of immunity required for vaccine development to combat pandemics like the COVID-19. url: https://doi.org/10.1101/2020.06.11.147363 doi: 10.1101/2020.06.11.147363 id: cord-320559-up1q3k6q author: Dortmans, J.C.F.M. title: Porcine epidemic diarrhea virus (PEDV) introduction into a naive Dutch pig population in 2014 date: 2018-05-24 words: 4216.0 sentences: 207.0 pages: flesch: 57.0 cache: ./cache/cord-320559-up1q3k6q.txt txt: ./txt/cord-320559-up1q3k6q.txt summary: Porcine epidemic diarrhea virus (PEDV) is the highly contagious, causative agent of an economically important acute enteric disease in pigs of all ages. In total, 838 blood samples from sows from 267 farms and 101 samples from wild boars were collected from May till November 2014 and tested for antibodies against PEDV by ELISA. The number of required blood samples from animals and farms to estimate the seroprevalence of PEDV in Dutch sow herds was calculated based on the following assumptions: PEDV is highly contagious and no vaccination against this virus was carried out in the Netherlands. For the detection of PEDV antibodies in serum samples an in-house indirect ELISA based on the viral spike (S) protein S1-part of the G2b strain GDU (Non-S-INDEL, GenBank KU985230.1) was used, similar as the ELISA previously described (Gerber et al., 2014) . abstract: Porcine epidemic diarrhea virus (PEDV) is the highly contagious, causative agent of an economically important acute enteric disease in pigs of all ages. The disease is characterized by diarrhea and dehydration causing mortality and growth retardation. In the last few decades, only classical PEDV was reported sporadically in Europe, but in 2014 outbreaks of PEDV were described in Germany. Phylogenetic analysis showed a very high nucleotide similarity with a variant of PEDV that was isolated in the US in January 2014. The epidemiological situation of PEDV infections in the Netherlands in 2014 was unknown and a seroprevalence study in swine was performed. In total, 838 blood samples from sows from 267 farms and 101 samples from wild boars were collected from May till November 2014 and tested for antibodies against PEDV by ELISA. The apparent herd prevalence of 0.75% suggests that PEDV was not circulating on a large scale in the Netherlands at this time. However, in November 2014 a clinical outbreak of PEDV was diagnosed in a fattener farm by PCR testing. This was the first confirmed PEDV outbreak since the early nineties. Sequence analyses showed that the viruses isolated in 2014 and 2015 in the Netherlands cluster with recently found European G1b strains. This suggests a one event introduction of PEDV G1b strains in Europe in 2014, which made the Netherlands and other European countries endemic for this type of strains since then. url: https://www.ncbi.nlm.nih.gov/pubmed/29981699/ doi: 10.1016/j.vetmic.2018.05.014 id: cord-254318-w8wrn9lx author: Díez, José-María title: Currently available intravenous immunoglobulin contains antibodies reacting against severe acute respiratory syndrome coronavirus 2 antigens date: 2020-05-13 words: 2687.0 sentences: 167.0 pages: flesch: 48.0 cache: ./cache/cord-254318-w8wrn9lx.txt txt: ./txt/cord-254318-w8wrn9lx.txt summary: MATERIAL & METHODS: Gamunex(®)-C and Flebogamma(®) DIF (Grifols) intravenous immunoglobulin (IVIG) products were tested using ELISA techniques for antibodies against several antigens of human common betacoronaviruses that may crossreact with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. Gamunex R -C (Grifols Therapeutics, Inc., NC, USA) and Flebogamma R DIF (Instituto Grifols S.A., Barcelona, Spain) IVIGs were tested for crossreactivity against several betacoronaviruses, including SARS-CoV, MERS-CoV and SARS-CoV-2 antigens, using ELISA techniques. Even with this uncertainty, in the context of the current health emergency (pandemic), the potential of IVIG as a therapy for COVID-19 is already being evaluated in a number of studies involving patients with severe SARS-CoV-2 viral infections including pneumonia [28] [29] [30] . • This is the first time that currently available intravenous immunoglobulins have been reported to contain antibodies that crossreact against antigens of SARS-CoV-2 and other coronaviruses. abstract: AIM: There is a critical need for effective therapies that are immediately available to control the spread of COVID-19 disease. MATERIAL & METHODS: Gamunex(®)-C and Flebogamma(®) DIF (Grifols) intravenous immunoglobulin (IVIG) products were tested using ELISA techniques for antibodies against several antigens of human common betacoronaviruses that may crossreact with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. RESULTS: Both IVIGs showed consistent reactivity to components of the tested viruses. Positive crossreactivity was seen in SARS-CoV, middle east respiratory syndrome-CoV and SARS-CoV-2. For SARS-CoV-2, positive reactivity was observed at IVIG concentrations ranging from 100 μg/ml with Gamunex-C to 1 mg/ml with Flebogamma 5% DIF. CONCLUSION: Gamunex-C and Flebogamma DIF contain antibodies reacting against SARS-CoV-2 antigens. Studies to confirm the utility of IVIG preparations for COVID-19 management may be warranted. url: https://www.ncbi.nlm.nih.gov/pubmed/32397847/ doi: 10.2217/imt-2020-0095 id: cord-252867-lku0cm62 author: Edwards, S. title: A comparison of three rapid diagnostic methods for the detection of rotavirus infection in calves date: 1987-01-31 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Abstract Three techniques for the detection of rotavirus in faecal samples from calves with neonatal gastroenteritis were compared. A preliminary study indicated that reverse passive haemagglutination (RPHA) was at least as sensitive as the enzyme-linked immunosorbent assay (ELISA). These two immunoassays were compared with the detection of viral RNA by polyacrylamide gel electrophoresis (PAGE) on 209 field samples. Of the 77 samples in which at least one test gave a positive result, 69 were positive by both RPHA and PAGE, but only 49 were also positive by ELISA, indicating a lower sensitivity for the latter test. The overall agreement between RPHA and PAGE was 96%. The reasons for the discrepancies between the tests are discussed. url: https://www.sciencedirect.com/science/article/pii/0378113587900940 doi: 10.1016/0378-1135(87)90094-0 id: cord-257190-iesysf3l author: Eshaghi, Majid title: Purification of the extra-cellular domain of Nipah virus glycoprotein produced in Escherichia coli and possible application in diagnosis date: 2005-03-30 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: The glycoprotein (G) of Nipah virus (NiV) is important for virus infectivity and induction of the protective immunity. In this study, the extra-cellular domain of NiV G protein was fused with hexahistidine residues at its N-terminal end and expressed in Escherichia coli. The expression under transcriptional regulation of T7 promoter yielded insoluble protein aggregates in the form of inclusion bodies. The inclusion bodies were solubilized with 8 M urea and the protein was purified to homogeneity under denaturing conditions using nickel–nitrilotriacetic acid (Ni–NTA) affinity chromatography. The denatured protein was renatured by gradual removal of the urea. Light scattering analysis of the purified protein showed primarily monodispersity. The purified protein showed significant reactivity with the antibodies present in the sera of NiV-infected swine, as demonstrated in Western blot analysis and enzyme-linked immunosorbent assay (ELISA). Taken together, the data indicate the potential usefulness of the purified G protein for structural or functional studies and the development of immunoassay for detection of the NiV antibodies. url: https://www.ncbi.nlm.nih.gov/pubmed/15707682/ doi: 10.1016/j.jbiotec.2004.10.020 id: cord-342024-kaku49xd author: Espejo, Andrea P title: Review of Current Advances in Serologic Testing for COVID-19 date: 2020-06-25 words: 6480.0 sentences: 373.0 pages: flesch: 50.0 cache: ./cache/cord-342024-kaku49xd.txt txt: ./txt/cord-342024-kaku49xd.txt summary: • The use of total antibody or simultaneous IgG/IgM measurements (regardless of method) significantly adds sensitivity to reverse transcription polymerase chain reaction testing protocols early post onset of symptoms and becomes the most accurate diagnostic test at later time points. The SP, RBD, and NP proteins appear to be the main targets of the humoral immune response in coronavirus infections including SARS-CoV-2 and were the antigens used in the majority of the serologic assays examined in this literature review. Overall, while these results are similar to that reported in a review of serologic testing for MERS-CoV and SARS-CoV, they may have been affected by choice of target antigens, the various immunoassay kits, and the level of detail of case history used to categorize the time of sample acquisition post onset of symptoms. abstract: OBJECTIVES: To examine and summarize the current literature on serologic methods for the detection of antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). METHODS: A literature review was performed using searches in databases including PubMed, medRxiv, and bioRxiv. Thirty-two peer-reviewed papers and 23 preprints were examined. RESULTS: The studies included lateral flow immunoassay, enzyme-linked immunosorbent assay, chemiluminescence immunoassay, and neutralizing antibody assays. The use of all major SARS-CoV-2 antigens was demonstrated to have diagnostic value. Assays measuring total antibody reactivity had the highest sensitivity. In addition, all the methods provided opportunities to characterize the humoral immune response by isotype. The combined use of IgM and IgG detection resulted in a higher sensitivity than that observed when detecting either isotype alone. Although IgA was rarely studied, it was also demonstrated to be a sensitive marker of infection, and levels correlated with disease severity and neutralizing activity. CONCLUSIONS: The use of serologic testing, in conjunction with reverse transcription polymerase chain reaction testing, was demonstrated to significantly increase the sensitivity of detection of patients infected with SARS-CoV-2. There was conflicting evidence regarding whether antibody titers correlated with clinical severity. However, preliminary investigations indicated some immunoassays may be a surrogate for the prediction of neutralizing antibody titers and the selection of recovered patients for convalescent serum donation. url: https://www.ncbi.nlm.nih.gov/pubmed/32583852/ doi: 10.1093/ajcp/aqaa112 id: cord-301355-9lswjro2 author: Fan, Jing-Hui title: Development of an enzyme-linked immunosorbent assay for the monitoring and surveillance of antibodies to porcine epidemic diarrhea virus based on a recombinant membrane protein date: 2015-12-01 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: The recent dramatic increase in reported cases of porcine epidemic diarrhea (PED) in pig farms is a potential threat to the global swine industry. Therefore, the accurate diagnosis, serological monitoring, and surveillance of specific antibodies in pigs resulting from porcine epidemic diarrhea virus (PEDV) infection or vaccination would be essential in helping to control the spread of PED. We developed and validated an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant membrane (M) protein of PEDV. To detect PEDV antibodies in eight herds, 382 serum samples were collected from sows that had been immunized with a PED vaccine, and screened using the developed ELISA in parallel with a serum neutralization (SN) assay. Of the tested samples, 276 were positive for the presence of PEDV antibodies according to both assays, while 98 were negative. An excellent agreement between the ELISA and the SN assay was observed (kappa = 0.947; 95% confidence interval = 0.910–0.984; McNemar's test, P = 0.727). No cross-reaction was detected for the developed ELISA with other coronaviruses or other common pig pathogens. The developed ELISA could be used for serological evaluation and indirect diagnosis of PED infection. url: https://api.elsevier.com/content/article/pii/S0166093415002645 doi: 10.1016/j.jviromet.2015.07.021 id: cord-310536-u30cufg7 author: Finger, Paula Fonseca title: Combined use of ELISA and Western blot with recombinant N protein is a powerful tool for the immunodiagnosis of avian infectious bronchitis date: 2018-12-12 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: The avian infectious bronchitis virus (IBV) remains a significant source of loss in the poultry industry and early diagnosis is required to prevent the disease from spreading. This study examined the combined use of an ELISA and Western blot (WB) to detect antibodies against the nucleocapsid protein (N) of IBV. The coding sequence for N was amplified by RT-PCR and expressed in Escherichia coli. A soluble recombinant N protein (rN) of approximately 50 kDa was obtained. A total of 389 sera were tested against the rN in ELISA and the results were compared with those of the commercial IDEXX IBV Ab test. ELISA-rN achieved a 90.34% sensitivity and 90.16% specificity. WB confirmed all false negative sera in ELISA-rN or IDEXX test as truly positive. The current study indicate that the combined use of rN in ELISA and WB is a powerful tool for the immunodiagnosis of avian infectious bronchitis. METHODS: Constructed recombinant pAE/n expression vectors were used to transform E. coli BL21(DE3) Star competent cells (Invitrogen). The rN of infectious bronchitis virus was purified by affinity chromatography using HisTrap HP 1 mL columns pre-packed with pre-charged Ni Sepharose in the ÄKTAprime Automated Liquid Chromatography system (GE Healthcare). A total of 389 serum samples from chickens were used to develop and evaluate the ELISA-rN test. To standardize the indirect ELISA development, serum dilutions (1:100, 1:200 and 1:400) and different concentrations of purified rN antigen (50, 100 and 200 ng/well) were tested. Positive and negative sera for IBV were used as controls. The results were compared with those obtained from a commercial kit. Serum samples scored as negative with the commercial kit but as positive with the ELISA-rN were further analysed by Western blot analyses using the rN protein as an antigen. The results of the ELISA-rN were compared to the commercial kit results using receiver-operating characteristics curves, area under the curve, and confidence intervals with the software GraphPad Prism version 6.0 for Windows (GraphPad Software, USA). RESULTS: The expected cDNA fragment of approximately 1240 bp was successfully amplified by PCR using primers designed to select for the coding region of the N protein. The rN was expressed as a soluble protein to avoid the refolding steps and, after purification a yield of 10 mg/L of rN was obtained. The SDS-PAGE results demonstrated the presence of two distinct bands that had a molecular mass of approximately 45 and 50 KDa. Out of 244 sera that scored positive in the commercial ELISA IDEXX IBV Ab Test, 220 were also positive in the ELISA-rN, yielding an ELISA-rN test sensitivity of 90.16%. Out of 145 sera that scored negative in the IDEXX IBV Ab Test, 131 also scored negative in the ELISA-rN, indicating a specificity of 90.34%. Sera that tested negative in the ELISA-rN and positive in the commercial test also reacted with the rN protein in Western blot. CONCLUSIONS: The association between the ELISA and Western blot techniques developed in this study with a subunit of IBV (rN) were able to detect antibodies that the commercial ELISA did not detect suggesting that the ELISA-rN has greater sensitivity. url: https://doi.org/10.1186/s12985-018-1096-2 doi: 10.1186/s12985-018-1096-2 id: cord-289413-mbrw85og author: Flego, Michela title: Intracellular human antibody fragments recognizing the VP35 protein of Zaire Ebola filovirus inhibit the protein activity date: 2019-09-05 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Ebola hemorrhagic fever is caused by the Ebola filovirus (EBOV), which is one of the most aggressive infectious agents known worldwide. The EBOV pathogenesis starts with uncontrolled viral replication and subversion of both the innate and adaptive host immune response. The multifunctional viral VP35 protein is involved in this process by exerting an antagonistic action against the early antiviral alpha/beta interferon (IFN-α/β) response, and represents a suitable target for the development of strategies to control EBOV infection. Phage display technology permits to select antibodies as single chain Fragment variable (scFv) from an artificial immune system, due to their ability to specifically recognize the antigen of interest. ScFv is ideal for genetic manipulation and to obtain antibody constructs useful for targeting either antigens expressed on cell surface or intracellular antigens if the scFv is expressed as intracellular antibody (intrabody) or delivered into the cells. RESULTS: Monoclonal antibodies (mAb) in scFv format specific for the EBOV VP35 were isolated from the ETH-2 library of human recombinant antibodies by phage display technology. Five different clones were identified by sequencing, produced in E.coli and expressed in CHO mammalian cells to be characterized in vitro. All the selected scFvs were able to react with recombinant VP35 protein in ELISA, one of the scFvs being also able to react in Western Blot assay (WB). In addition, all scFvs were expressed in cell cytoplasm as intrabodies; a luciferase reporter gene inhibition assay performed in A549 cells showed that two of the scFvs can significantly hamper the inhibition of the IFN-β-induced RIG-I signaling cascade mediated by EBOV VP35. CONCLUSION: Five antibodies in scFv format recognize an active form of EBOV VP35 in ELISA, while one antibody also recognizes VP35 in WB. Two of these scFvs were also able to interfere with the intracellular activity of VP35 in a cell system in vitro. These findings suggest that such antibodies in scFv format might be employed to develop therapeutic molecules able to hamper EBOV infections. url: https://doi.org/10.1186/s12896-019-0554-2 doi: 10.1186/s12896-019-0554-2 id: cord-313517-5ipj2z86 author: Fung, Joshua title: Antigen Capture Enzyme-Linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans date: 2019-09-14 words: 2710.0 sentences: 149.0 pages: flesch: 51.0 cache: ./cache/cord-313517-5ipj2z86.txt txt: ./txt/cord-313517-5ipj2z86.txt summary: Though the gold standard for diagnosing MERS-CoV infection in humans is still nucleic acid amplification test (NAAT) of the up-E region, an antigen capture enzyme-linked immunosorbent assay (ELISA) could also be of use for early diagnosis in less developed locations. In the present method, a step-by-step guide to perform a MERS-CoV nucleocapsid protein (NP) capture ELISA using two NP-specific monoclonal antibodies is provided for readers to develop their in-house workflow or diagnostic kit for clinical use and for mass-screening project of animals (e.g., dromedaries and bats) to better understand the spread and evolution of the virus. Nucleic acid amplification test (NAAT, e.g., real-time reverse transcription quantitative polymerase chain reaction [real-time RT-qPCR]), virus isolation, transmission electron microscopy, immunohistochemistry, and serological methods (e.g., antigen capture enzyme-linked immunosorbent assay [ELISA] and immunofluorescence assay [IFA] ) have been developed and used for MERS-CoV diagnosis [2] [3] [4] [5] [6] [7] . abstract: The Middle East respiratory syndrome (MERS) is the second novel zoonotic disease infecting humans caused by coronavirus (CoV) in this century. To date, more than 2200 laboratory-confirmed human cases have been identified in 27 countries, and more than 800 MERS-CoV associated deaths have been reported since its outbreak in 2012. Rapid laboratory diagnosis of MERS-CoV is the key to successful containment and prevention of the spread of infection. Though the gold standard for diagnosing MERS-CoV infection in humans is still nucleic acid amplification test (NAAT) of the up-E region, an antigen capture enzyme-linked immunosorbent assay (ELISA) could also be of use for early diagnosis in less developed locations. In the present method, a step-by-step guide to perform a MERS-CoV nucleocapsid protein (NP) capture ELISA using two NP-specific monoclonal antibodies is provided for readers to develop their in-house workflow or diagnostic kit for clinical use and for mass-screening project of animals (e.g., dromedaries and bats) to better understand the spread and evolution of the virus. url: https://doi.org/10.1007/978-1-0716-0211-9_7 doi: 10.1007/978-1-0716-0211-9_7 id: cord-003471-tr3ageky author: Gaikwad, Satish S. title: Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein date: 2019-01-31 words: 3398.0 sentences: 199.0 pages: flesch: 51.0 cache: ./cache/cord-003471-tr3ageky.txt txt: ./txt/cord-003471-tr3ageky.txt summary: PURPOSE: The aim of the present study was to develop a serodiagnostic test for differentiation infected from vaccinated animal (DIVA) strategy accompanying the marker vaccine lacking an immunodominant epitope (IDE) of nucleoprotein of Newcastle disease virus (NDV). Epitope repeat protein (ERP) gene encoding eight repeats of an IDE sequence containing ETQFLDLMRAVANSMR (C-terminal IDE aa 444-459) on NDV NP separated by tetra-glycine linker was synthesized commercially using codon optimization for baculovirus expression. In addition to SPF chicken serum, different hyper-immune chicken sera to NDV, H9N2 avian influenza virus (AIV), infectious bronchitis virus (IBV), and infectious bursal disease virus (IBDV) were tested for specificity of rERP using an indirect ELISA assay. Second and third groups were sera taken 14 dpi from SPF chickens vaccinated with IDE deleted recombinant Newcastle disease virus (rNDV) [4] and NDV LaSota strain [19] , respectively, which kindly supplied by the OIE reference laboratory for ND, Korea. abstract: PURPOSE: The aim of the present study was to develop a serodiagnostic test for differentiation infected from vaccinated animal (DIVA) strategy accompanying the marker vaccine lacking an immunodominant epitope (IDE) of nucleoprotein of Newcastle disease virus (NDV). MATERIALS AND METHODS: Recombinant epitope-repeat protein (rERP) gene encoding eight repeats of the IDE sequence (ETQFLDLMRAVANSMR) by tetra-glycine linker was synthesized. Recombinant baculovirus carrying the rERP gene was generated to express the rERP in insect cells. Specificity and sensitivity of an indirect enzyme-linked immunosorbent assay (ELISA) employing the rERP was evaluated. RESULTS: The rERP with molecular weight of 20 kDa was successfully expressed by the recombinant baculovirus in an insect-baculovirus system. The rERP was antigenically functional as demonstrated by Western blotting. An indirect ELISA employing the rERP was developed and its specificity and sensitivity was determined. The ELISA test allowed discrimination of NDV infected sera from epitope deletion virus vaccinated sera. CONCLUSION: The preliminary results represent rERP ELISA as a promising DIVA diagnostic tool. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6369128/ doi: 10.7774/cevr.2019.8.1.27 id: cord-007497-nn5l5rai author: Garcia-Sanchez, J. title: Survey of rotavirus infection in a dairy herd: comparison between polycrylamide gel electrophoresis and two commercial tests date: 2002-11-13 words: 3578.0 sentences: 165.0 pages: flesch: 54.0 cache: ./cache/cord-007497-nn5l5rai.txt txt: ./txt/cord-007497-nn5l5rai.txt summary: Faecal samples taken from 15 cows before and after calving as well as faeces taken from their calves daily from birth to two weeks of life were tested for rotavirus by polyacrylamide gel electrophoresis (PAGE) and compared with an ELISA and a latex agglutination commercial test. Various methods have been developed for rapid detection of rotaviruses in faecal samples; these include electron microscopy (considered for a long time as the reference method), immune electron microscopy, counter immuno-electrophoresis, complement fixation, fluorescent antibody tests, different immunoassays using antibodies labelled with radioisotopes or enzymes (ELISA), latex agglutination and polyacrylamide gel electrophoresis (PAGE). All the faeces samples (120 from cows and 240 from calves) were tested for the presence of rotaviruses by polyacrylamide gel electrophoresis (PAGE), a commercial ELISA kit and a commercial latex agglutination kit. Another three discrepancies were detected in which the samples were positive by PAGE and negative by both ELISA and latex agglutination and all of them corresponded to two calves with a discontinuous shedding pattern (calf no. abstract: A survey of rotavirus infection in a dairy herd with a history of neonatal diarrhoea was carried out. Faecal samples taken from 15 cows before and after calving as well as faeces taken from their calves daily from birth to two weeks of life were tested for rotavirus by polyacrylamide gel electrophoresis (PAGE) and compared with an ELISA and a latex agglutination commercial test. Rotavirus excretion was not detected in faeces from cows around parturition by any of the three tests. However, all of their calves shed rotaviruses during the observation period. The onset of rotavirus excretion determined by PAGE ranged from day 2 to day 8 of life (day 4.8± 1.8 on average) and lasted for 4 to 7 days (5.3±1.1 days on average). Chi-square test showed a significant association (P=0.0001) between the presence of rotavirus and the altered consistency of calves faeces. All the three tests showed similar results (overall agreement 92.5%) but discrepancies were detected mainly at the beginning or at the end of the rotavirus excretion period. Results obtained with both commercial kits closely paralleled each other and parameters other than sensitivity, specificity, diagnostic accuracy or predictive values have to be considered as selection criteria. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117447/ doi: 10.1016/0378-1135(93)90057-e id: cord-013348-lsksys56 author: Goto, Keiko title: Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 date: 2020-09-19 words: 4742.0 sentences: 247.0 pages: flesch: 50.0 cache: ./cache/cord-013348-lsksys56.txt txt: ./txt/cord-013348-lsksys56.txt summary: title: Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3 From the resultant hybridoma clones, we selected nine clones producing mAbs reactive to the HPeV3-VP0 antigen, based on enzyme-linked immunosorbent assay (ELISA). Although RT-PCR-based diagnostic tests targeting 5 -UTR of the HPeV3 genome were developed for HPeV3 detection in clinical samples, there is currently no diagnostic method for detecting the viral antigens. To develop a rapid and effective diagnostic strategy, there is an urgent need to produce highly specific monoclonal antibodies (mAbs) toward HPeV3 antigens. As a result, we obtained nine mAb clones for characterization, and thereafter, generated an ELISA system that is specifically able to detect the HPeV3 VP0 antigens. We next performed antigen-capture ELISA with recombinant VP0 protein and virions released into the cell-culture supernatant of the HPeV3-infected cells. In this study, we sought to generate specific mAbs and develop an ELISA test for the detection of HPeV3 VP0 antigen. abstract: Human parechovirus type 3 (HPeV3) is an etiologic agent of respiratory diseases, meningitis, and sepsis-like illness in both infants and adults. Monoclonal antibodies (mAbs) can be a promising diagnostic tool for antigenic diseases such as virus infection, as they offer a high specificity toward a specific viral antigen. However, to date, there is no specific mAb available for the diagnosis of HPeV3 infection. In this study, we developed and characterized mAbs specific for HPeV3 capsid protein VP0. We used cell-free, wheat germ-synthesized viral VP0 protein for immunizing BALB/c mice to generate hybridomas. From the resultant hybridoma clones, we selected nine clones producing mAbs reactive to the HPeV3-VP0 antigen, based on enzyme-linked immunosorbent assay (ELISA). Epitope mapping showed that these mAbs recognized three distinct domains in HPeV3 VP0. Six mAbs recognized HPeV3 specifically and the other three mAbs showed cross-reactivity with other HPeVs. Using the HPeV3-specific mAbs, we then developed an ELISA for viral antigen detection that could be reliably used for laboratory diagnosis of HPeV3. This ELISA system exhibited no cross-reactivity with other related viruses. Our newly developed mAbs would, thus, provide a useful set of tools for future research and ensure HPeV3-specific diagnosis. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7563955/ doi: 10.3390/microorganisms8091437 id: cord-285700-9q6vwoct author: Grzelak, Ludivine title: SARS-CoV-2 serological analysis of COVID-19 hospitalized patients, pauci-symptomatic individuals and blood donors. date: 2020-04-24 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: It is of paramount importance to evaluate the prevalence of both asymptomatic and symptomatic cases of SARS-CoV-2 infection and their antibody response profile. Here, we performed a pilot study to assess the levels of anti-SARS-CoV-2 antibodies in samples taken from 491 pre- epidemic individuals, 51 patients from Hopital Bichat (Paris), 209 pauci-symptomatic individuals in the French Oise region and 200 contemporary Oise blood donors. Two in-house ELISA assays, that recognize the full-length nucleoprotein (N) or trimeric Spike (S) ectodomain were implemented. We also developed two novel assays: the S-Flow assay, which is based on the recognition of S at the cell surface by flow-cytometry, and the LIPS assay that recognizes diverse antigens (including S1 or N C- terminal domain) by immunoprecipitation. Overall, the results obtained with the four assays were similar, with differences in sensitivity that can be attributed to the technique and the antigen in use. High antibody titers were associated with neutralisation activity, assessed using infectious SARS-CoV- 2 or lentiviral-S pseudotypes. In hospitalized patients, seroconversion and neutralisation occurred on 5-14 days post symptom onset, confirming previous studies. Seropositivity was detected in 29% of pauci-symptomatic individuals within 15 days post-symptoms and 3 % of blood of healthy donors collected in the area of a cluster of COVID cases. Altogether, our assays allow for a broad evaluation of SARS-CoV2 seroprevalence and antibody profiling in different population subsets. url: http://medrxiv.org/cgi/content/short/2020.04.21.20068858v1?rss=1 doi: 10.1101/2020.04.21.20068858 id: cord-015683-a9a82of4 author: Gupta, Varsha title: Molecular Diagnostics date: 2016-10-23 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Effective and early management of diseases requires record of the history, behavioral parameters, and travel information. These are helpful for the diagnosis, prevention, and control of the disease. There have been several advancements in the methods for diagnosing infectious diseases. The wide spectrum of tests such as biochemical evaluation, microbiological tools, immunological and molecular biology techniques, etc., is available. Each type of diagnostic technique is strong and reliable in its own sense but poses certain limitations. These limitations may be complemented by using a combination of tests. Older techniques such as microscopy and culturing of organisms from clinical specimens are error-free but are very labor intensive and extremely time consuming. There is a need to develop rapid and sensitive tests that can be used in both high- and low-resource settings. Molecular diagnostics such as Western blot, ELISA, PCR, DNA, and protein microarrays are revolutionizing the clinical practice of infectious diseases. Their effects are significant in acute-care settings where timely and accurate diagnostic tools are critical for patient treatment decisions and outcomes. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7115026/ doi: 10.1007/978-981-10-0875-7_9 id: cord-279406-wwdqh9qs author: Guzman, Norberto A. title: A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy date: 2020-07-30 words: 17172.0 sentences: 835.0 pages: flesch: 33.0 cache: ./cache/cord-279406-wwdqh9qs.txt txt: ./txt/cord-279406-wwdqh9qs.txt summary: To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses. It would be beneficial to have a sample processing method before separation, to isolate and concentrate the intended viruses or EVs. Immunoaffinity capillary electrophoresis has already been proven to be a useful technology to isolate, separate, and quantify cell-free molecules of biological interest based on the specificity and selectivity not only of antibody reagents, but also of lectin and aptamer reagents, quantifying molecules ranging from microgram/milliliter to femtogram/milliliter [25, 54, 55, 57, 75] . abstract: Biomarker detection for disease diagnosis, prognosis, and therapeutic response is becoming increasingly reliable and accessible. Particularly, the identification of circulating cell-free chemical and biochemical substances, cellular and subcellular entities, and extracellular vesicles has demonstrated promising applications in understanding the physiologic and pathologic conditions of an individual. Traditionally, tissue biopsy has been the gold standard for the diagnosis of many diseases, especially cancer. More recently, liquid biopsy for biomarker detection has emerged as a non-invasive or minimally invasive and less costly method for diagnosis of both cancerous and non-cancerous diseases, while also offering information on the progression or improvement of disease. Unfortunately, the standardization of analytical methods to isolate and quantify circulating cells and extracellular vesicles, as well as their extracted biochemical constituents, is still cumbersome, time-consuming, and expensive. To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. Isolation and concentration of analytes is accomplished through binding to one or more biorecognition affinity ligands immobilized to a solid support, while separation and analysis are achieved by high-resolution capillary electrophoresis (CE) coupled to one or more detectors. When compared to other existing methods, the process of this affinity capture, enrichment, release, and separation of one or a panel of biomarkers can be carried out on-line with the advantages of being rapid, automated, and cost-effective. Additionally, it has the potential to demonstrate high analytical sensitivity, specificity, and selectivity. As the potential of liquid biopsy grows, so too does the demand for technical advances. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses. url: https://doi.org/10.3390/biomedicines8080255 doi: 10.3390/biomedicines8080255 id: cord-343185-lbmbp9ca author: Hansen, C. B. title: SARS-CoV-2 antibody responses determine disease severity in COVID-19 infected individuals date: 2020-07-29 words: 5349.0 sentences: 332.0 pages: flesch: 51.0 cache: ./cache/cord-343185-lbmbp9ca.txt txt: ./txt/cord-343185-lbmbp9ca.txt summary: Here we have developed novel flexible ELISA-based assays for specific detection of SARS-CoV-2 antibodies against the receptor-binding domain (RBD): An antigen sandwich-ELISA relevant for large population screening and three isotype-specific assays for in-depth diagnostics. Detection of IgM, IgA and IgG antibodies against SARS-CoV-2 protein N was evaluated by analyzing 136 positive samples and 174 negative controls and ROC curve analyses were assessed to estimate the assay performance . To provide a better insight into antibody seroconversion during SARS-CoV-2 infection and reactivity against different locations on protein S and protein N, we conducted IgM, IgA and IgG detection in 90 positive samples against 14 protein fragments and short peptides located on the protein S and protein N structures, full-length RBD, protein S and protein N (Figure 2A ). We have developed an ELISA-based platform for detection SARS-CoV-2 antibodies comprising an indirect RBD S-ELISA for pan Ig detection and direct ELISAs for in-depth analyses of the IgM, IgA and IgG isotype responses towards RBD and protein N. abstract: Globally, the COVID-19 pandemic has had extreme consequences for the healthcare system and calls for diagnostic tools to monitor and understand the transmission, pathogenesis and epidemiology, as well as to evaluate future vaccination strategies. Here we have developed novel flexible ELISA-based assays for specific detection of SARS-CoV-2 antibodies against the receptor-binding domain (RBD): An antigen sandwich-ELISA relevant for large population screening and three isotype-specific assays for in-depth diagnostics. Their performance was evaluated in a cohort of 350 convalescent participants with previous COVID-19 infection, ranging from asymptomatic to critical cases. We mapped the antibody responses to different areas on protein N and S and showed that the IgM, A and G antibody responses against RBD are significantly correlated to the disease severity. These assays-and the data generated from them-are highly relevant for diagnostics and prognostics and contribute to the understanding of long-term COVID-19 immunity. url: http://medrxiv.org/cgi/content/short/2020.07.27.20162321v1?rss=1 doi: 10.1101/2020.07.27.20162321 id: cord-277186-sj8ngpk8 author: He, Qigai title: Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA date: 2005-04-19 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: This report describes the production of several MAbs against N195 protein, a major immunodomain of SARS CoV nucleocapsid protein [He, Q., Chong, K.H., Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004. Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. Clin. Diagn. Lab. Immunol. 11 (2) 417–422.]. One representative IgG1 monoclonal antibody (MAb), S-A5D5, was selected and characterized. S-A5D5 reacted specifically react with both recombinant and native nucleocapsid protein of SARS CoV. The reactivity of S-A5D5 with purified N195 protein and utilization of the MAb as a detector antibody to develop an antigen capture ELISA was assessed. As little as 37.5 pg of purified N protein and 50 TCID(50) of SARS CoV could be detected by the antigen capture ELISA. Specific binding of the MAb S-A5D5 to both purified N195 and SARS CoV nucleocapsid antigen was effectively inhibited by human SARS positive serum and guinea pig anti-N195 serum. The N protein in N195-spike recombinant baculovirus-infected Sf-9 cells could also be identified. N protein was detected in 18 IFA IgM-positive serum samples collected from SARS confirmed patients, but not in nine samples collected from SARS recovery patient. No false positive results were given when 60 samples from healthy individuals were tested, and no cross-reaction occurred when infectious bronchitis virus (IBV), chicken coronavirus, was tested. This monoclonal antibody-based antigen capture ELISA is thus a powerful tool for early diagnosis of SARS CoV infection. url: https://api.elsevier.com/content/article/pii/S0166093405000856 doi: 10.1016/j.jviromet.2005.03.004 id: cord-010247-cug21fnf author: Hollingshead, Melinda G. title: An ELISA system for evaluating antiretroviral activity against Rauscher murine leukemia virus date: 1992-06-15 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: A system for evaluating the activity of antiviral agents against Rauscher murine leukemia virus (R-MuLV) has been developed using an enzyme linked immunosorbent assay technique. The activity of various antiviral compounds demonstrated in this assay system has been compared to their activity in the UV-XC plaque reduction assay, which has been used historically for evaluating anti-R-MuLV compounds. The assay is based upon detection of R-MuLV encoded p30 protein production in virus infected murine cells. The assay reagents are readily available and the assay system is amenable to automated data collection systems. Cytotoxicity evaluations are conducted in parallel to the Rauscher MuLV ELISA assay in order to assess drug-induced reductions in cell viability. Cytotoxicity evaluations are important to interpretation of the ELISA results since reductions in cell viability reduce viral protein production which would indicate an antiviral drug effect. This system is less sensitive than the classical UV-XC plaque reduction assay; however, it does offer an alternative to the time-consuming and labor-intensive plaque assay. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7173104/ doi: 10.1016/0166-3542(92)90060-i id: cord-331277-fjsuo3yy author: Hoste, Alexis C.R. title: Two serological approaches for detection of antibodies to SARS-CoV-2 in different scenarios: A screening tool and a point-of-care test date: 2020-08-11 words: 2407.0 sentences: 136.0 pages: flesch: 52.0 cache: ./cache/cord-331277-fjsuo3yy.txt txt: ./txt/cord-331277-fjsuo3yy.txt summary: Two serological tools based on a Double Recognition assay (Enzyme-Linked Immunosorbent Assay, DR-ELISA and Lateral Flow Assay, DR-LFA) to detect total antibodies to SARS-CoV-2, have been developed based on the recombinant nucleocapsid protein. Therefore, the aim of the present work was the development of serological tools to determine the presence of antibodies against SARS-CoV-2 in the population, as an indicator of an ongoing or previous infection. In the current study, a Double Recognition Enzyme-Linked Immunosorbent Assay (DR-ELISA) was developed to determine the presence of immunoglobulins of different classes (IgG, IgM and IgA) to SARS-CoV-2 in human serum, to support the diagnosis of COVID-19. In the present study, we developed two serological assays using the recombinant N protein Table 1, a group of 14 serum samples from early days post infection, positive to COVID-19 by respiratory-PCR yet still negative in the commercial serological assay (with seroconversion a few days later) were also tested in our assays. abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has infected more than 8 million people worldwide, becoming a pandemic. Detecting antibodies against SARS-CoV-2 is of utmost importance and a good indicator of exposure and circulation of the virus within the general population. Two serological tools based on a Double Recognition assay (Enzyme-Linked Immunosorbent Assay, DR-ELISA and Lateral Flow Assay, DR-LFA) to detect total antibodies to SARS-CoV-2, have been developed based on the recombinant nucleocapsid protein. A total of 1065 serum samples, including positive for COVID-19 and negative samples from healthy donors or infected with other respiratory pathogens, were analyzed. The results showed values of sensitivity between 91.2%–100%, and specificity of 100%–98.2%, for DR-LFA and DR-ELISA, respectively. No cross-reactivity against seasonal coronavirus (HCoV-NL63, HCoV-229E, HCoV-HKU1, HCoV-OC43) was found. These results demonstrate the importance of serology as a complementary tool to PCR, for follow up of recovered patients and identification of asymptomatic individuals. url: https://doi.org/10.1016/j.diagmicrobio.2020.115167 doi: 10.1016/j.diagmicrobio.2020.115167 id: cord-321691-46la29tm author: Hsueh, Po-Ren title: SARS Antibody Test for Serosurveillance date: 2004-09-17 words: 2800.0 sentences: 133.0 pages: flesch: 44.0 cache: ./cache/cord-321691-46la29tm.txt txt: ./txt/cord-321691-46la29tm.txt summary: A peptide-based enzyme-linked immunosorbent assay (ELISA) can be used for retrospective serosurveillance of severe acute respiratory syndrome (SARS) by helping identify undetected chains of disease transmission. A peptide-based enzyme-linked immunosorbent assay (ELISA) can be used for retrospective serosurveillance of severe acute respiratory syndrome (SARS) by helping identify undetected chains of disease transmission. Such surveillance may be key to tracking the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) because mild and asymptomatic cases of SARS-CoV infection that do not meet the World Health Organization''s case definition (1) have been identified by immunoassays (2) (3) (4) , and SARS-CoV-like viruses have been isolated from wild mammals (5) . The diagnostic sensitivity of the peptide ELISA was 100% on a panel of 69 convalescent-phase serum samples from SARS patients provided as a reference panel by the Center for Disease Control, Department of Health, Taiwan. The peptide ELISA was evaluated for specificity on serum samples drawn from patients associated with typical and atypical respiratory pathogens other than SARS-CoV (National Taiwan University Hospital). abstract: A peptide-based enzyme-linked immunosorbent assay (ELISA) can be used for retrospective serosurveillance of severe acute respiratory syndrome (SARS) by helping identify undetected chains of disease transmission. The assay was developed by epitope mapping, using synthetic peptides from the spike, membrane, and nucleocapsid protein sequences of SARS-associated coronavirus. The new peptide ELISA consistently detected seroconversion by week 2 of onset of fever, and seropositivity remained through day 100. Specificity was 100% on normal blood donor samples, on serum samples associated with infection by other pathogens, and on an interference panel. The peptide-based test has advantages of safety, standardization, and automation over previous immunoassays for SARS. The assay was used for a retrospective survey of healthy healthcare workers in Taiwan who treated SARS patients. Asymptomatic seroconversions were detected in two hospitals that had nosocomial disease. url: https://www.ncbi.nlm.nih.gov/pubmed/15498156/ doi: 10.3201/eid1009.040101 id: cord-300701-vkzya7uq author: Ijaz, M. K. title: Effect of different routes of immunization with bovine rotavirus on lactogenic antibody response in mice date: 1987-12-31 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Summary The effect of different routes of immunization with either live or killed bovine rotavirus (BRV) on the production of lactogenic antibody response in mice was evaluated. The routes of immunization were intramuscular (IM), oral (O) or intradermal in the mammary region (IMam). Following immunization, serum antibody responses were monitored by an enzyme linked immunosorbent assay (ELISA). Following whelping, the mice were allowed to stay with their mother until sacrificed on alternate days post-parturition from day 1–11. Milk from their stomach was collected for antibody titration by ELISA and virus neutralization test. Regardless of the routes of immunization, a rapid increase in serum anti-rotavirus antibody titers was observed for the first 5 wk after immunization followed by a gradual decline. After parturition, the mean antibody titer of lacteal secretions, as determined by ELISA, increased gradually for 7 days with the greatest increase on day 9, followed by a decrease in anti-rotavirus antibody. These titers also correlated with antibody titers in milk as measured by virus neutralization test. The best lactogenic antibody response was observed when IMam × IM × 2 route of immunization was used with live BRV as the antigen. Interestingly, immunization via the oral route with killed BRV also resulted in good antibody responses. In contrast, in the group where killed BRV was used, animals receiving 3× orally had the highest antibody titer. The distribution of different antibody subtypes in milk samples revealed IgG to be the predominant antibody followed by IgM and IgA. Irrespective of the route of administration, there was an increase in IgA on day 9 as compared to day 1 in most of the groups. The significant role played by mucosal immunity in passive protection and the possible ways to modulate subtype specific lactogenic immune response are discussed. Animals models; Lactogenic immunity; Rotaviruses url: https://www.ncbi.nlm.nih.gov/pubmed/2837143/ doi: 10.1016/s0166-3542(87)80006-2 id: cord-335121-ro3x3qa3 author: Ingram, George A. title: A comparative assessment of four serological methods used in the detection and measurement of anti-parasite antibodies in the serum of the amphibian, bufo viridis date: 1988-04-30 words: 3116.0 sentences: 180.0 pages: flesch: 50.0 cache: ./cache/cord-335121-ro3x3qa3.txt txt: ./txt/cord-335121-ro3x3qa3.txt summary: Abstract Antibodies against Crithidia fasciculata choanomastigotes were detected in green toad (Bufo viridis) sera by direct agglutination, indirect haemagglutination (IHA), complement-fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA), Correlation coefficients (r) were calculated for comparisons between each of the techniques and regression formulae derived in order to convert antibody levels as determined by one immunological method to that of another. In this paper we present the results of acomparative assessment of four serological tests (direct agglutination, indirect haemagglutination, complement-fixation test and ELISA) used to detect and to determine the levels of antibodies in the sera of green toads (B. fasciculata and the number of control and immune sera containing detectable antibodies were the lowest for DA and CFT, intermediate for IHA and highest for the ELISA method (Table 1) . Nevertheless the method of antigen preparation ma> not be a salient criterion for antibody estimation in immune sera because correlation values of over 89% (f''< 0.001) were found when IHA titres were compared to those of ELISA. abstract: Abstract Antibodies against Crithidia fasciculata choanomastigotes were detected in green toad (Bufo viridis) sera by direct agglutination, indirect haemagglutination (IHA), complement-fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA), Correlation coefficients (r) were calculated for comparisons between each of the techniques and regression formulae derived in order to convert antibody levels as determined by one immunological method to that of another. The highest mean titre obtained by ELISA was approximately 1.5–3.5 times greater than those obtained by the other techniques whilst CFT gave the lowest values. IHA and ELISA titres were affected by different preparations of the crithidial antigen extracts. Highly significant r values were determined for control sera when IHA was compared to ELISA (r > 0.79), and to both CFT and ELISA with immune animals (r > 0.96). ELISA would seem most applicable for screening other lower vertebrates for anti-parasite antibodies especially in areas of human disease prevalence. url: https://api.elsevier.com/content/article/pii/0020751988901476 doi: 10.1016/0020-7519(88)90147-6 id: cord-003315-r1wkx0ml author: Jacobs, Sophie title: Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ date: 2018-11-01 words: 6190.0 sentences: 337.0 pages: flesch: 59.0 cache: ./cache/cord-003315-r1wkx0ml.txt txt: ./txt/cord-003315-r1wkx0ml.txt summary: We next developed a firefly luciferase-based reporter cell line, named Fawa-λ-luc, to detect IFN-λ in biological fluids with high specificity and sensitivity. This bioassay was as sensitive as a commercially available enzyme-linked immunosorbent assay in detecting mouse IFN-λ in cell culture supernatant, as well as in serum and bronchoalveolar lavage samples of virus-infected mice. Unlike mouse and human type I IFNs that exhibit strongly species-specific activity (Veomett and Veomett 1979) , type III IFNs were reported to act on cell types from both origins (Lasfar and others 2006; Hermant and others 2014) . IFNl4 was quantified by a cytopathic effect reduction assay in A549 cells, using recombinant human IFN-l3 as a standard, which was reported to have a similar specific activity (Hamming and others 2013). We then compared the sensitivities of the Fawa-l-luc and of the ELISA assays to detect IFN-l in mouse serum and BAL samples. abstract: The type III interferon (IFN-λ) family includes 4 IFN-λ subtypes in man. In the mouse, only the genes coding for IFN-λ2 and -λ3 are present. Unlike mouse and human type I IFNs (IFN-α/β), which exhibit strong species specificity, type III IFNs were reported to act in a cross-specific manner. We reexamined the cross-specificity and observed that mouse and human IFN-λ exhibit some species specificity, although much less than type I IFNs. Mouse IFN-λ3 displayed clear species specificity, being 25-fold less active in human cells than the closely related mouse IFN-λ2. This specificity likely depends on amino acids in α helices A and F that diverged from other IFN-λ sequences. Human IFN-λ4, in contrast, retained high activity in mouse cells. We next developed a firefly luciferase-based reporter cell line, named Fawa-λ-luc, to detect IFN-λ in biological fluids with high specificity and sensitivity. Fawa-λ-luc cells, derived from mouse epithelial cells that are responsive to IFN-λ, were made nonresponsive to type I IFNs by inactivation of the Ifnar2 gene and strongly responsive to IFN-λ by overexpression of the mouse IFNLR1. This bioassay was as sensitive as a commercially available enzyme-linked immunosorbent assay in detecting mouse IFN-λ in cell culture supernatant, as well as in serum and bronchoalveolar lavage samples of virus-infected mice. The assay also enabled the sensitive detection of human IFN-λ activity, including that of the divergent IFN-λ4 with a bias, however, due to variable activity of IFN-λ subtypes. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249671/ doi: 10.1089/jir.2018.0066 id: cord-255390-dvp0luxe author: Jones, R. D title: Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles date: 2000-04-10 words: 4834.0 sentences: 237.0 pages: flesch: 39.0 cache: ./cache/cord-255390-dvp0luxe.txt txt: ./txt/cord-255390-dvp0luxe.txt summary: Abstract The purpose of this subacute 22-day study was to evaluate methods for canine circulating immunoglobulins (IgM, IgG, and IgE) and select Band T-lymphocyte populations (CD4-helpers, CD8-suppressors, pan-T and pan-B) for immunotoxicity testing using an organ system (concordance) approach. The flow cytometry method was acceptable for measuring select canine lymphocyte populations and detecting the expected decrease in B cells due to cyclophosphamide treatment. The purpose of this study was to evaluate immunoglobulins (IgM, IgG, IgE) by ELISA and lymphocytes (CD4-helpers, CD8-suppressors, pan-T CD5 and pan-B CD21) by flow cytometry for detecting adverse effects in dogs administered positive control challenges to the immune system. The next day following each immunization, serum samples were taken for analysis of IgG, IgM and IgE, along with clinical chemistry, hematology, urinalysis, and flow cytometry on day 8, to evaluate possible influences of repeated immunization. abstract: Abstract The purpose of this subacute 22-day study was to evaluate methods for canine circulating immunoglobulins (IgM, IgG, and IgE) and select B- and T-lymphocyte populations (CD4-helpers, CD8-suppressors, pan-T and pan-B) for immunotoxicity testing using an organ system (concordance) approach. The challenge substance for immunoglobulin testing was repeated immunization with six-way distemper vaccination (DHLAPP), while the challenge substance for leukocyte subpopulations was treatment with cyclophosphamide. Immunoglobulin measurements were made by capture enzyme-linked immunosorbent assay (ELISA), and leukocyte immunophenotyping by fluorescein isothiocyanate/phycoerythrin conjugation (flow cytometry). A battery of parameters that would be used in a typical regulatory study were taken to aid interpretation of the data generated by these methods. Body weights, food consumption, clinical observations, complete clinical chemistry and urinalysis measurements were taken. Gross pathology and micropathology of sternal bone marrow, spleen, mesenteric and retropharyngeal lymph nodes, thymus, liver and kidney were completed. The ELISA method demonstrated acceptable intra-assay reproducibility for IgM, IgG and IgE, with values in good agreement as reported for radial immunodiffusion. The immunologic challenge demonstrated a biological trend of an increase in IgM that preceded an increase in IgG with no discernible trend in IgE response, and no abnormalities in lymphocyte subpopulations. Principle flow cytometry findings related to cyclophosphamide were that the relative percent of B cells decreased dramatically and progressively after compound administration; being statistically decreased in males on day 22 compared with day −5. The relative percent CD4 and CD8 contribution increased, but the CD4/CD8 ratio remained relatively unchanged as total white blood cells decreased progressively. The increase in relative percent CD4 (males only) was statistically significant according to a two-sample t-test on days 17, 20 and 22 when compared with the pre-treatment day −5. There was a relative percent increase in CD5-panT, but absolute numbers were dramatically decreased. We conclude that an organ system approach to assessment of the immune system which incorporates humoral antibody, enumeration of lymphocyte populations and pathologic evaluation of the lymphoreticular organs assists in the interpretation of an adverse toxicological response. The ELISA method for measurement of Igs detected the expected levels of IgG, IgM and IgE due to repeated vaccinations and to cyclophosphamide treatment. The flow cytometry method was acceptable for measuring select canine lymphocyte populations and detecting the expected decrease in B cells due to cyclophosphamide treatment. Both methods may be added to a testing battery for assessing immunotoxicity in canine regulatory studies. url: https://www.sciencedirect.com/science/article/pii/S0378427400001739 doi: 10.1016/s0378-4274(00)00173-9 id: cord-336899-zngp67tb author: Kadkhoda, Kamran title: COVID‐19: are neutralizing antibodies neutralizing enough? date: 2020-06-03 words: 1211.0 sentences: 73.0 pages: flesch: 51.0 cache: ./cache/cord-336899-zngp67tb.txt txt: ./txt/cord-336899-zngp67tb.txt summary: The use of convalescent-phase plasma (CP) in severely ill patients with COVID-19 has been attempted on an individual basis 1 and it is being used in the context of ongoing clinical trials; thus, it is pivotal to understand the potential risks and caveats of using CP particularly taking immunopathologic phenomena into account. In this context, previous exposure to common coronaviruses would lead to an early and high-titer immune response to SARS-CoV-2. The donors had neutralizing antibody titers of more than 640 at the time of donation while severely ill patients had relatively similar titers before transfusion as high as 640 (range, 160-640; GMT, 367). 7 The US Food and Drug Administration currently recommends using donated plasma with neutralizing antibody titers of 160 or at a minimum a titer of 80 in severely ill patients. abstract: nan url: https://www.ncbi.nlm.nih.gov/pubmed/32449171/ doi: 10.1111/trf.15897 id: cord-267736-rya9w6sh author: Kang, Xiaoping title: Development of an ELISA-array for simultaneous detection of five encephalitis viruses date: 2012-02-27 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Japanese encephalitis virus(JEV), tick-borne encephalitis virus(TBEV), and eastern equine encephalitis virus (EEEV) can cause symptoms of encephalitis. Establishment of accurate and easy methods by which to detect these viruses is essential for the prevention and treatment of associated infectious diseases. Currently, there are still no multiple antigen detection methods available clinically. An ELISA-array, which detects multiple antigens, is easy to handle, and inexpensive, has enormous potential in pathogen detection. An ELISA-array method for the simultaneous detection of five encephalitis viruses was developed in this study. Seven monoclonal antibodies against five encephalitis-associated viruses were prepared and used for development of the ELISA-array. The ELISA-array assay is based on a "sandwich" ELISA format and consists of viral antibodies printed directly on 96-well microtiter plates, allowing for direct detection of 5 viruses. The developed ELISA-array proved to have similar specificity and higher sensitivity compared with the conventional ELISAs. This method was validated by different viral cultures and three chicken eggs inoculated with infected patient serum. The results demonstrated that the developed ELISA-array is sensitive and easy to use, which would have potential for clinical use. url: https://www.ncbi.nlm.nih.gov/pubmed/22369052/ doi: 10.1186/1743-422x-9-56 id: cord-294478-3ickafd3 author: Kapil, Sanjay title: Diagnostic Investigation of Emerging Viruses of Companion Animals date: 2008-05-22 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: In this article, the authors are specifically concerned with the timely and accurate detection of emerging diseases of small animals that are viral in origin. Veterinarians are bound to encounter emerging viruses in their practice. The problem is unavoidable, because viruses are highly mutagenic. Even the immune response dictates the nature of virus that evolves in a host. If the clinical signs and diagnostic methods fail to correlate, the veterinarian should work with the diagnostic laboratory to solve the diagnostic puzzle. url: https://www.ncbi.nlm.nih.gov/pubmed/18501276/ doi: 10.1016/j.cvsm.2008.02.009 id: cord-333026-9f6ecg30 author: Kompanikova, J. title: Microbiologic Methods in the Diagnostics of Upper Respiratory Tract Pathogens date: 2017-03-03 words: 1784.0 sentences: 100.0 pages: flesch: 49.0 cache: ./cache/cord-333026-9f6ecg30.txt txt: ./txt/cord-333026-9f6ecg30.txt summary: Blood samples were simultaneously examined by the enzyme-linked immunosorbent assay (ELISA) and by the FilmArray Respiratory Panel for eight different pathogens in a total of 15 tests performed in nasopharyngeal swabs. Nonetheless, since most repiratory tract infections are viral in origin and there is no treatment available, the diagnosis provided by the FilmArray Panel does not provide any additional clinical benefit and thus should be used only whenever necessary on the individual basis. This method allows for identification of 21 different respiratory pathogens from a nasopharyngeal swab, 18 of viral etiology and three of bacterial origin (Idaho Technology 2007). The laboratory costs to run one examination with different methods showed that the FilmArray multiplex PCR respiratory panel is more expensive than the ELISA, HIT, and the cultivation. Since most URIs are viral in origin and there is no treatment available, the diagnosis provided by the FilmArray panel is not always necessary and the method should be used on an individual basis when clinically justified. abstract: Upper respiratory tract infection (URI) is a nonspecific term used to describe acute infections involving the nose, paranasal sinuses, pharynx, and larynx above the vocal cords. The aim of this study was to provide a summary of the most common pathogens of URI and to compare advantages and disadvantages of traditional and new rapid microbiological tests used to identify them. Blood samples were simultaneously examined by the enzyme-linked immunosorbent assay (ELISA) and by the FilmArray Respiratory Panel for eight different pathogens in a total of 15 tests performed in nasopharyngeal swabs. The ELISA method is unable to identify the pathologic agent until the host’s immune system elicits a response. The method is readily available in many laboratories at a low cost, which puts less strain on economic resources. The FilmArray(®) Panel, on the other hand, is more expensive, but it is fast and exact in the identification of a broad spectrum etiologic agents. Nonetheless, since most repiratory tract infections are viral in origin and there is no treatment available, the diagnosis provided by the FilmArray Panel does not provide any additional clinical benefit and thus should be used only whenever necessary on the individual basis. url: https://doi.org/10.1007/5584_2017_10 doi: 10.1007/5584_2017_10 id: cord-007480-grndfx7b author: Koopmans, M. title: Seroepidemiology of Breda virus in cattle using ELISA date: 2002-11-13 words: 3354.0 sentences: 176.0 pages: flesch: 55.0 cache: ./cache/cord-007480-grndfx7b.txt txt: ./txt/cord-007480-grndfx7b.txt summary: Two direct blocking enzyme linked immunosorbent assays (ELISA) for the detection of antibodies to Breda virus in sera of cattle were compared. This "Breda virus" (BRV) is morphologically and antigenically different from known bovine viruses and causes diarrhea in gnotobiotic and colostrum-deprived calves (Woode et al., 1982) . In the first approach (consecutive method) a 1:400 dilution of the BRV2 antigen preparation, purified from calf feces was added to the coated wells in ELISA buffer 1 (PBS supplemented with 0.35 M NaC1, 1 mM EDTA pH 7.5 and 0.05% Tween 80 ). When using the consecutive method more animals were found negative for antibodies to BRV2 or had low blocking percentages (Table 1 ) . In Fig. 3 , the percentages of animals with antibodies to BRV ( >~ 20% blocking, corresponding to a serum titer of/> 30 ) are shown for different age groups (n--244). abstract: Two direct blocking enzyme linked immunosorbent assays (ELISA) for the detection of antibodies to Breda virus in sera of cattle were compared. An ELISA with consecutive addition of antigen and test serum to an antibody-coated plate gave higher positive: negative absorbence ratios than an ELISA in which antigen and test serum were added simultaneously. Sera collected from breeding and fattening herds in The Netherlands (n = 1313) and the F.R.G. (n = 716) were tested, and antibodies to Breda virus were demonstrated in 94% of adult cattle. Ninety percent of newborn calves had high levels of maternal antibodies, which waned until the age of 3 months. Active seroconversion occurred between 7 and 24 months in most animals. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117312/ doi: 10.1016/0378-1135(89)90069-2 id: cord-025922-84pheilu author: Kostopoulou, Despoina title: Mapping the canine vector-borne disease risk in a Mediterranean area date: 2020-06-03 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: The aim of this study was to determine exposure to vector-borne pathogens (VBPs) in populations of dogs living on Greek islands in the Ionian and Aegean seas. METHODS: In total, 1154 dogs with different lifestyles and of varying ages and breeds were randomly sampled and examined for the presence of clinical signs compatible with canine vector-borne diseases (CVBDs). Blood was collected from each individual animal. For the detection of antibodies against Leishmania spp., the WITNESS® Leishmania test was performed, and positive samples were further examined with indirect enzymatic immunoassay (ELISA). Antibodies to Borrelia burgdorferi, Ehrlichia canis or E. ewingii, as well as Anaplasma phagocytophilum or A. platys were investigated using the Snap® 4Dx® Plus test. Positive Ehrlichia spp. and Anaplasma spp. samples were further examined using an indirect ELISA for further identification of the species. RESULTS: In total, 25.6% of dogs were exposed to at least one of the pathogens investigated, with seroprevalences varying regionally. Of these seropositive dogs, 27.4% displayed clinical signs suggestive of CVBDs, such as cutaneous lesions, enlarged lymph nodes, pale mucous membranes, onychogryphosis and weight loss. The overall seroprevalence detected using the rapid tests was 15.3% for Leishmania spp., whereas 2.3% of the examined dogs were found to be positive for Anaplasma spp. and 7.5% for Ehrlichia spp. while B. burgdorferi was not detected. Twenty-four samples positive to A. phagocytophilum by ELISA were analysed by PCR for the presence of Anaplasma DNA. PCR and sequencing results showed the presence of A. platys DNA in 4 samples and E. canis DNA in 4 samples. The remaining samples (66.7%) were negative. CONCLUSIONS: In the present study, exposure of dogs to VBPs was shown in the geographical areas investigated. Results confirm that on Greek islands VBPs represent a constant health risk for both native and visiting dogs, suggesting the presence of distinct “hot-spots” of VBP infections on different islands. In order to reduce the risk of transmission and the spread to non-endemic regions, the protection of dogs through use of repellents and vaccines, together with owner education, seem to be of paramount importance. [Image: see text] url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7268178/ doi: 10.1186/s13071-020-04153-8 id: cord-334277-g3go3u02 author: Kovac, Marc title: EDTA-Anticoagulated Whole Blood for SARS-CoV-2 Antibody Testing by Electrochemiluminescence Immunoassay (ECLIA) and Enzyme-Linked Immunosorbent Assay (ELISA) date: 2020-08-14 words: 4725.0 sentences: 237.0 pages: flesch: 53.0 cache: ./cache/cord-334277-g3go3u02.txt txt: ./txt/cord-334277-g3go3u02.txt summary: While lateral flow test formats can be utilized with whole blood and low sample volumes, their diagnostic characteristics are inferior to immunoassays based on chemiluminescence immunoassay (CLIA) or enzyme-linked immunosorbent assay (ELISA) technology. We addressed the suitability of EDTA-anticoagulated whole blood as an alternative sample material for antibody testing against SARS-CoV-2 by electro-CLIA (ECLIA; Roche, Rotkreuz, Switzerland) and ELISA (IgG and IgA; Euroimmun, Germany). In receiver-operating characteristic curve analysis, all three assays displayed comparable diagnostic accuracy (area under the curve (AUC)) using corrected whole blood and serum (AUCs: 0.97 for ECLIA and IgG ELISA; 0.84 for IgA ELISA). It can thus be concluded that the anti-SARS-CoV-2 antibody results in whole blood corrected for hematocrit with weakly and moderately positive findings are comparable to those obtained from serum. abstract: While lateral flow test formats can be utilized with whole blood and low sample volumes, their diagnostic characteristics are inferior to immunoassays based on chemiluminescence immunoassay (CLIA) or enzyme-linked immunosorbent assay (ELISA) technology. CLIAs and ELISAs can be automated to a high degree but commonly require larger serum or plasma volumes for sample processing. We addressed the suitability of EDTA-anticoagulated whole blood as an alternative sample material for antibody testing against SARS-CoV-2 by electro-CLIA (ECLIA; Roche, Rotkreuz, Switzerland) and ELISA (IgG and IgA; Euroimmun, Germany). Simultaneously drawn venous serum and EDTA-anticoagulated whole blood samples from 223 individuals were included. Correction of the whole blood results for hematocrit led to a good agreement with the serum results for weakly to moderately positive antibody signals. In receiver-operating characteristic curve analysis, all three assays displayed comparable diagnostic accuracy (area under the curve (AUC)) using corrected whole blood and serum (AUCs: 0.97 for ECLIA and IgG ELISA; 0.84 for IgA ELISA). In conclusion, our results suggest that the investigated assays can reliably detect antibodies against SARS-CoV-2 in hemolyzed whole blood anticoagulated with EDTA. Correction of these results for hematocrit is suggested. This study demonstrates that the automated processing of whole blood for identification of SARS-CoV-2 antibodies with common ECLIA and ELISA methods is accurate and feasible. url: https://www.ncbi.nlm.nih.gov/pubmed/32823852/ doi: 10.3390/diagnostics10080593 id: cord-353614-z4fpy607 author: Kusano, Nario title: Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies date: 2007-02-16 words: 2976.0 sentences: 147.0 pages: flesch: 54.0 cache: ./cache/cord-353614-z4fpy607.txt txt: ./txt/cord-353614-z4fpy607.txt summary: title: Immunochromatographic assay for simple and rapid detection of Satsuma dwarf virus and related viruses using monoclonal antibodies A simple and rapid immunochromatographic assay (ICA) to detect Satsuma dwarf virus (SDV) was developed using colloidal gold conjugates of anti-SDV monoclonal antibodies. Comparison of the sensitivities of ICA and DAS-ELISA Out of the three combinations of using three Mabs for ICA, clone 2G2 and clone 4E6, which had a different paratope group (PG), were more sensitive and suitable to the colloidal gold conjugate and coating antibody on the nitrocellulose strip for detection of SDV, respectively (data not shown). Suspensions of colloidal gold conjugate at various concentrations were applied to the reagent pad, and ICA The immunochromatographic devices were prepared under the following conditions: Case 3-1, 1 mg/ml of anti-SDV polyclonal antibody was deposited onto a nitrocellulose membrane to make a test line. abstract: A simple and rapid immunochromatographic assay (ICA) to detect Satsuma dwarf virus (SDV) was developed using colloidal gold conjugates of anti-SDV monoclonal antibodies. Of six homogenization buffers tested, 0.1 M citrate buffer (pH 7.0) gave the best results for the ICA. In the ICA, addition of 0.1% thioglycolic acid in the homogenization buffers that have been widely used in enzyme-linked immunosorbent assays (ELISA) was deleterious to the reaction because of undesirable coagulation of the colloidal gold. ICA using the anti-SDV monoclonal antibodies was 8 times and 16 times more sensitive than double antibody sandwich-ELISA and ICA using the anti-SDV polyclonal antibody, respectively. The analysis is complete in only 15 min. Furthermore, ICA using the anti-SDV monoclonal antibodies could also detect SDV-related viruses. url: https://www.ncbi.nlm.nih.gov/pubmed/32214869/ doi: 10.1007/s10327-006-0316-6 id: cord-022310-yc6xtw0s author: Lappin, Michael R. title: Microbiology and Infectious Disease date: 2011-12-15 words: 14109.0 sentences: 913.0 pages: flesch: 39.0 cache: ./cache/cord-022310-yc6xtw0s.txt txt: ./txt/cord-022310-yc6xtw0s.txt summary: 47 Because of these findings, it is currently recommended to combine serologic test results with those of blood culture or PCR assay results when evaluating clinically ill cats for bartonellosis. Because serologic test results do not accurately correlate with presence of bacteremia and individual culture or PCR assay results can be falsely negative, there is no indication for testing healthy cats for Bartonella spp. T. gondii-specific IgM is detectable in serum by ELISA in approximately 80% of subclinically ill cats 2 to 4 weeks after experimental induction of toxoplasmosis; these titers generally are negative less than 16 weeks after infection. The author, however, has demonstrated IgG antibody titers greater than 1 : 16,384 in subclinically ill cats 5 years after experimental induction In chronic disease or asymptomatic carriers, demonstration of organisms is unreliable, and a tentative diagnosis is based on clinical signs and a positive titer. gondii-specific antibodies can also be detected in the serum, CSF, and aqueous humor of healthy, infected animals, one cannot assay results in clinically ill dogs, E. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7155555/ doi: 10.1016/b978-1-4377-0657-4.00015-6 id: cord-349744-8cg5yj20 author: Lassaunière, Ria title: Evaluation of nine commercial SARS-CoV-2 immunoassays date: 2020-04-10 words: 2649.0 sentences: 145.0 pages: flesch: 53.0 cache: ./cache/cord-349744-8cg5yj20.txt txt: ./txt/cord-349744-8cg5yj20.txt summary: The results showed 100% specificity for the Wantai SARS-CoV-2 Total Antibody ELISA, 93% for the Euroimmun IgA ELISA, and 96% for the Euroimmun IgG ELISA with sensitivities of 90%, 90%, and 65%, respectively. While the four POC tests evaluated according to illness duration were often weakly positive or detected only IgG or IgM during the early phase (data not shown), their sensitivities were comparable to the Wantai Total Ab ELISA and Euroimmun IgA ELISA in all three phases. In the present study, three SARS-CoV-2-specific commercial ELISA assays and six POC rapid tests were evaluated using sera from hospitalized adult patients with PCR-confirmed diagnoses for SARS-CoV-2 and a collection of control serum samples taken before the emergence of the virus in China in December 2019. Overall, the Wantai Total Ab ELISA had superior sensitivity and specificity compared to both Euroimmun IgA and IgG ELISAs. The POC tests varied notably, with the best performance observed for the test produced by AutoBio Diagnostics, followed by the tests produced by Dynamiker Biotechnology and CTK Biotech. abstract: Due to urgency and demand, numerous severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoassays are rapidly being developed and placed on the market with limited validation on clinical samples. Thorough validation of serological tests are required to facilitate their use in the accurate diagnosis of SARS-CoV-2 infection, confirmation of molecular results, contact tracing, and epidemiological studies. This study evaluated the sensitivity and specificity of nine commercially available serological tests. These included three enzyme-linked immunosorbent assays (ELISAs) and six point-of-care (POC) lateral flow tests. The assays were validated using serum samples from: i) SARS-CoV-2 PCR-positive patients with a documented first day of disease; ii) archived sera obtained from healthy individuals before the emergence of SARS-CoV-2 in China; iii) sera from patients with acute viral respiratory tract infections caused by other coronaviruses or non-coronaviruses; and iv) sera from patients positive for dengue virus, cytomegalovirus and Epstein Barr virus. The results showed 100% specificity for the Wantai SARS-CoV-2 Total Antibody ELISA, 93% for the Euroimmun IgA ELISA, and 96% for the Euroimmun IgG ELISA with sensitivities of 90%, 90%, and 65%, respectively. The overall performance of the POC tests according to manufacturer were in the rank order of AutoBio Diagnostics > Dynamiker Biotechnology = CTK Biotech > Artron Laboratories > Acro Biotech ≥ Hangzhou Alltest Biotech. Overall, these findings will facilitate selection of serological assays for the detection SARS-CoV-2-specific antibodies towards diagnosis as well as sero-epidemiological and vaccine development studies. url: https://doi.org/10.1101/2020.04.09.20056325 doi: 10.1101/2020.04.09.20056325 id: cord-301655-6nxhvvm4 author: Lei, Xi-Mei title: Specific recombinant proteins of porcine epidemic diarrhea virus are immunogenic, revealing their potential use as diagnostic markers date: 2019-08-10 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Given the highly contagious and acute nature of porcine epidemic diarrhea (PED), especially in piglets, there is an urgent need for the development of rapid and sensitive diagnostic assays. The diagnostic potentials of specific porcine epidemic diarrhea virus (PEDV) accessory and nonstructural proteins, if any, have not yet been investigated. In order to determine and compare which of the viral proteins may be useful as diagnostic antigens, whole virus (WV) particles and a panel of structural and nonstructural PEDV proteins [spike subunit 1 (S1), the C-terminal part of ORF3 (ORF3C), envelope (E), nonstructural protein 1 (Nsp1), Nsp2, Ac (acidic domain of Nsp3), and ADRP (ADP-ribose-1-monophosphatase domain of Nsp3), expressed individually in bacterial and/or mammalian cells] were tested for reactivity with sera from PEDV-infected pigs by ELISA and/or western blot analysis. According to western blots, serum antibody interactions with the S1 protein were relatively more sensitive and specific than ORF3C, E and Ac. Furthermore, a total of 851 serum samples from diarrheal pigs of different ages were analyzed by ELISA, with most showing immune-reactivity towards the WV, S1, ORF3C, and E proteins. The earliest IgG antibody response was observed in the one-week-old piglets, with similar antibody ontogeny and patterns of seroconversion for S1, ORF3C, E, and WV antigens. In addition, the pattern of neutralizing antibody was more similar to that of IgA in weaning piglets after PEDV infection. Collectively, these data provide more reliable information on the host immune response to different viral proteins, which will be useful for development of novel serological assays and for design of vaccines that better stimulate protective immunity. url: https://www.sciencedirect.com/science/article/pii/S0378113519304365 doi: 10.1016/j.vetmic.2019.108387 id: cord-291104-6chpmgry author: Leung, Danny T. M. title: Osteopontin Fragments with Intact Thrombin-Sensitive Site Circulate in Cervical Cancer Patients date: 2016-08-05 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: We investigated whether circulating osteopontin (OPN) could be used as a biomarker for cervical cancer. We employed a monoclonal antibody (mAb 659) specific for the unique and intact thrombin-sensitive site in OPN using an inhibition ELISA. We found significantly higher levels of OPN in 33 cervical cancer patients in both the plasma (mean +/- SD, 612 +/- 106 ng/mL) and serum (424 +/- 121 ng/mL) compared to healthy subjects [409 +/- 56 ng/mL, from 31 plasma samples (P < 0.0001), and 314 +/- 98 ng/mL, from 32 serum samples (P = 0.0002), respectively]. Similar results were obtained when the plasma from a bigger group (147 individuals) of cervical cancer patients (560 +/- 211 ng/mL) were compared with the same plasma samples of the healthy individuals (P = 0.0014). More significantly, the OPN level was highest in stage III-IV disease (614 +/- 210 ng/mL, from 52 individuals; P = 0.0001) and least and non-discriminatory in stage I (473 +/- 110 ng/mL, from 40 individuals; P = 0.5318). No such discrimination was found when a mAb of a different specificity (mAb 446) was used in a similar inhibition ELISA to compare the two groups in the first study; a commercial capture ELISA also failed. The possibility that the target epitope recognized by the antibody probe in these assays was absent from the circulating OPN due to protein truncation was supported by gel fractionation of the OPN found in patients’ plasma: 60–64 kDa fragments were found instead of the presumably full-length OPN (68 kDa) seen in healthy people. How these fragments are generated and what possible role they play in cancer biology remain interesting questions. url: https://doi.org/10.1371/journal.pone.0160412 doi: 10.1371/journal.pone.0160412 id: cord-277735-a9gkath5 author: Leung, Danny Tze Ming title: Antibody Response of Patients with Severe Acute Respiratory Syndrome (SARS) Targets the Viral Nucleocapsid date: 2004-07-15 words: 4010.0 sentences: 192.0 pages: flesch: 55.0 cache: ./cache/cord-277735-a9gkath5.txt txt: ./txt/cord-277735-a9gkath5.txt summary: We examined serum samples obtained from 46 patients with SARS, 40 patients with non-SARS pneumonia, and 38 healthy individuals, by use of Western blotting (WB), enzyme-linked immunoassay (ELISA), and immunofluorescence assay, using both native and bacterially produced antigens of the virus. Both the humoral and cellular arms of the adaptive immune response are presumed to be important in controlling 2 or preparation U reacted with a serum sample from a patient with SARS showing the highly reactive antigens-nucleocapsid (N) 1, N2, and N3-in the former. Second, we made a recombinant antigen of the N-terminal half of the N protein (rNa), and, when it was used in an IgG ELISA, we found results almost identical to those found with the crude viral extract (89% sensitivity and 94%-95% specificity), including 4 negative cases in common ( figure 2) . abstract: The recent outbreak of severe acute respiratory syndrome (SARS) provided an opportunity to study the antibody response of infected individuals to the causative virus, SARS coronavirus. We examined serum samples obtained from 46 patients with SARS, 40 patients with non-SARS pneumonia, and 38 healthy individuals, by use of Western blotting (WB), enzyme-linked immunoassay (ELISA), and immunofluorescence assay, using both native and bacterially produced antigens of the virus. We found a highly restricted, immunoglobulin G-dominated antibody response in patients with SARS, directed most frequently (89% by ELISA) and predominantly at the nucleocapsid. Almost all of the subjects without SARS had no antinucleocapsid antibodies. The spike protein was the next most frequently targeted, but only 63% of the patients (by ELISA) responded. Other targets of the response identified by use of WB included antigens of 80 and 60 kDa. Several nonstructural proteins cloned were not antigenic, and the culture-derived nucleocapsid appeared to be specifically degraded. url: https://www.ncbi.nlm.nih.gov/pubmed/15216476/ doi: 10.1086/422040 id: cord-344309-6c2wttxg author: Lin, Huixing title: Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein date: 2018-08-20 words: 4411.0 sentences: 221.0 pages: flesch: 55.0 cache: ./cache/cord-344309-6c2wttxg.txt txt: ./txt/cord-344309-6c2wttxg.txt summary: title: Development and application of an indirect ELISA for the detection of antibodies to porcine epidemic diarrhea virus based on a recombinant spike protein In this study, an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant truncated spike (S) protein of PEDV was developed and validated. This indirect ELISA was compared to indirect immunoinfluscent assay (IFA), and the overall coincidence rate was 96.74% based on testing 368 clinical serum samples with different PEDV antibody levels. Finally, the S1 indirect ELISA was applied to detect serum antibodies of 3304 field samples collected from different pig farms in eastern China, and it presented an overall substantial agreement on the PEDV infection status. Therefore, this study selected a gene fragment within the S1 subunit as a coating antigen to develop an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of PEDV antibodies. Detection of antibodies against porcine epidemic diarrhea virus in serum and colostrum by indirect ELISA abstract: BACKGROUND: As the major causative agent of swine viral diarrhea, porcine epidemic diarrhea virus (PEDV) has caused massive losses to the economies of swine raising countries. Accordingly, the serological detection of corresponding antibodies would be beneficial to diagnose PEDV indirectly to control the disease. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant truncated spike (S) protein of PEDV was developed and validated. RESULTS: The reaction conditions of the developed indirect ELISA were optimized. This indirect ELISA was compared to indirect immunoinfluscent assay (IFA), and the overall coincidence rate was 96.74% based on testing 368 clinical serum samples with different PEDV antibody levels. No cross-reactivity with other common swine pathogens was detected for the developed S1 indirect ELISA. Finally, the S1 indirect ELISA was applied to detect serum antibodies of 3304 field samples collected from different pig farms in eastern China, and it presented an overall substantial agreement on the PEDV infection status. CONCLUSIONS: This established S1 indirect ELISA is capable of detecting serum antibodies against PEDV, and due to its high sensitivity and specificity, it could be applied for serological evaluation and indirect diagnosis of PEDV infection. url: https://doi.org/10.1186/s12917-018-1570-5 doi: 10.1186/s12917-018-1570-5 id: cord-276989-441aclcc author: Liu, Jianbo title: Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum date: 2020-03-12 words: 4023.0 sentences: 213.0 pages: flesch: 54.0 cache: ./cache/cord-276989-441aclcc.txt txt: ./txt/cord-276989-441aclcc.txt summary: title: Development of a rapid immunochromatographic strip test for the detection of porcine epidemic diarrhea virus specific SIgA in colostrum On the strip, a gold-labeled anti-SIgA SC mAb was applied to a conjugate pad; purified PEDV particles and goat anti-mouse antibodies were blotted onto a nitrocellulose membrane to form the test and control lines, respectively. Therefore, the aim of this study was to develop a simple and rapid immunochromatographic test strip incorporating a colloidal gold-labeled anti-SIgA SC mAb probe for the detection of anti-PEDV-specific SIgA in swine, and to compare its performance with an indirect SIgA ELISA based on the whole PEDV virus (Cong et al., 2019) . To test for colostrum, we diluted the samples 1: 20 in sample buffer and mixed thoroughly then, 100 μL of the solution was dispensed onto the sample pad well of the strip apparatus to determine the presence of PEDV specific SIgA, as described above. abstract: Porcine epidemic diarrhea virus (PEDV) causes very high mortality in newborn piglets. The mucosal immune system in the gut must eliminate potential pathogens while maintaining a mutually beneficial relationship with the commensal microbiota. Antibodies derived from the secretory immunoglobulin A (SIgA) class, act as the first line of antigen-specific immunity in the gut by recognizing both pathogens and commensals. Therefore, the measurement of SIgA levels is an important index in evaluating PEDV infections and immune status. A simple and rapid method for the detection of PEDV-specific SIgA using an immunochromatographic test strip has been developed; incorporating a colloidal gold-labeled anti-SIgA secretory component (SC) mAb probe for the detection of anti-PEDV-specific SIgA in swine. On the strip, a gold-labeled anti-SIgA SC mAb was applied to a conjugate pad; purified PEDV particles and goat anti-mouse antibodies were blotted onto a nitrocellulose membrane to form the test and control lines, respectively. Results showed that the immunochromatographic test strip had high sensitivity and specificity. When compared with enzyme-linked immunosorbent assay, kappa value suggesting that the strip could be used to detect PEDV specific SIgA in colostrum samples. Furthermore, the strip assay is rapid and easy to perform with no requirement for professional-level skills or equipment. We found that the immunochromatographic test strip was a rapid, sensitive, and reliable method for the identification of PEDV specific SIgA, indicating its suitability for epidemiological surveillance as well as vaccine immunity when studying PEDV. url: https://api.elsevier.com/content/article/pii/S0166093420301075 doi: 10.1016/j.jviromet.2020.113855 id: cord-011840-neowfhwg author: Liu, Weixiao title: Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops date: 2020-03-05 words: 3920.0 sentences: 232.0 pages: flesch: 52.0 cache: ./cache/cord-011840-neowfhwg.txt txt: ./txt/cord-011840-neowfhwg.txt summary: title: Development of a sensitive monoclonal antibody-based sandwich ELISA to detect Vip3Aa in genetically modified crops OBJECTIVES: To develop a sensitive monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) to detect Vip3Aa in genetically modified (GM) crops and their products. A sensitive monoclonal antibody-based sandwich ELISA was developed to detect Vip3Aa in GM crops and their products. These antibodies were then employed to develop a sandwich ELISA for sensitive, direct and convenient measurement of the Vip3Aa concentration in GM crops and their products. To the best of our knowledge, this is the first quantitative monoclonal antibody-based ELISA method reported for the sensitive detection of Vip3Aa proteins. In this study, we describe a sensitive monoclonal antibody-based ELISA method for the detection of Vip3Aa in GM crops and their products. Development of monoclonal antibodies against Cry1Ab protein from Bacillus thuringiensis and their application in an ELISA for detection of transgenic Bt-maize abstract: OBJECTIVES: To develop a sensitive monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) to detect Vip3Aa in genetically modified (GM) crops and their products. RESULTS: Vegetative insecticidal proteins (Vips) are secreted by Bacillus thuringiensis (Bt) and are known to be toxic to Lepidoptera species. Vip3Aa family proteins, Vip3Aa19 and Vip3Aa20, were successfully applied in GM crops to confer an effective and persistent insecticidal resistance. A sensitive monoclonal antibody-based sandwich ELISA was developed to detect Vip3Aa in GM crops and their products. Two monoclonal antibodies were raised against the overexpressed and purified His-Vip3Aa20, were purified from mouse ascites and characterized. A sandwich ELISA method was developed using the 2G3-1D7 monoclonal antibody for capture and the biotin-labeled 1F9-1F5 monoclonal antibody for detection of Vip3Aa20. The linear detection range of the method was found to be approximately 31.25–500 pg/ml, with a sensitivity of 10.24 pg/ml. CONCLUSIONS: The established ELISA was effective for detecting Vip3Aa family proteins other than Vip3Aa8, and was successfully applied in the detection of Vip3Aa20 and Vip3Aa19 expressed in transgenic maize and cotton. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7354279/ doi: 10.1007/s10529-020-02854-9 id: cord-254384-mwzz1db5 author: Lu, Guilan title: Large-scale seroprevalence analysis of human metapneumovirus and human respiratory syncytial virus infections in Beijing, China date: 2011-02-10 words: 3978.0 sentences: 242.0 pages: flesch: 57.0 cache: ./cache/cord-254384-mwzz1db5.txt txt: ./txt/cord-254384-mwzz1db5.txt summary: To assess the seroprevalence of hMPV infection in China, we tested a total of 1,156 serum specimens for the presence of anti-hMPV IgG antibody in children and adults free of acute respiratory illness in Beijing, China by using hMPV nucleocapsid (N) protein as an antigen. To assess the seroprevalence of hMPV infection in China, we used hMPV N protein as an antigen to test serum samples for the presence of anti-hMPV IgG antibody in children and adults free of acute respiratory illness in Beijing, China. Lower seropositive rates and geometric mean titer (GMT) of anti-hMPV IgG were observed in children aged six months to six years when compared to hRSV. To test the specificity of the ELISA methods established in this study, the reactions of mouse sera against influenza virus A (subtypes H1-H16), human coronaviruses (229E, HKU1 and NL63), and polyomavirus JC against hMPV and hRSV N protein were evaluated. abstract: BACKGROUND: Human metapneumovirus (hMPV), a recently identified virus, causes acute respiratory tract infections (ARTIs) in infants and children. However, studies on the seroepidemeology of hMPV are very limited in China. To assess the seroprevalence of hMPV infection in China, we tested a total of 1,156 serum specimens for the presence of anti-hMPV IgG antibody in children and adults free of acute respiratory illness in Beijing, China by using hMPV nucleocapsid (N) protein as an antigen. As a control, we used the human serum antibody against the N protein of human respiratory syncytial virus (hRSV), the most important viral agent responsible for ARIs in children. RESULTS: The seropositive rate for hMPV increased steadily with age from 67% at 1-6 mo to 100% at age 20. However, the rate dropped slightly between 6 mo and 1 yr of age. The seropositive rate for hRSV also increased steadily with age from 71% at 1-6 mo to 100% at age 20. In children aged six months to six years, the seropositive rates for the anti-hRSV IgG antibody were significantly higher than those for hMPV. Additionally, IgG antibody titers to hMPV and hRSV were significantly higher in adults than in young children. Consistent with the seropositive rates, the geometric mean titer of anti-hMPV IgG antibody was lower than that of anti-hRSV IgG antibody in children aged six months to six years. CONCLUSIONS: Our results indicate that similar to hRSV, exposure to hMPV is ubiquitous in the Beijing population. However, the seroprevalence of anti-hMPV IgG antibody is lower than that of hRSV in children between six months and six years old, which suggests a different number of repeat infections or a different response to infections. url: https://doi.org/10.1186/1743-422x-8-62 doi: 10.1186/1743-422x-8-62 id: cord-334896-3g75spkc author: MINAMI, Shohei title: Establishment of serological test to detect antibody against ferret coronavirus date: 2016-03-03 words: 2493.0 sentences: 150.0 pages: flesch: 59.0 cache: ./cache/cord-334896-3g75spkc.txt txt: ./txt/cord-334896-3g75spkc.txt summary: Since there is no available serological methods to detect antibodies to ferret coronavirus (FRCoV), an enzyme-linked immunosorbent assay (ELISA) using recombinant partial nucleocapsid (N) proteins of the ferret coronavirus (FRCoV) Yamaguchi-1 strain was developed to establish a serological method for detection of FRCoV infection. This different reactivity was also confirmed by immunoblot analysis using the serum from the ferret.Therefore, the a.a. 1–179 of the N protein was used as an ELISA antigen. In this study, an enzyme-linked immunosorbent assay (ELISA) using recombinant N proteins was established and applied to investigate the seroprevalence of FRcoV infection in Japan. The plasma of No.10 and serum of ferret No.22 showed differ-ent reactivities from the other samples in ELISA (Fig. 2) . In this study, we attempted to clarify the seroprevalence of FRcoV in Japan and developed an ELISA using two Yamaguchi-1 strain recombinant N proteins, GST-N (1-179) and GST-N (180-374). abstract: Since there is no available serological methods to detect antibodies to ferret coronavirus (FRCoV), an enzyme-linked immunosorbent assay (ELISA) using recombinant partial nucleocapsid (N) proteins of the ferret coronavirus (FRCoV) Yamaguchi-1 strain was developed to establish a serological method for detection of FRCoV infection. Many serum samples collected from ferrets recognized both a.a. 1–179 and a.a. 180–374 of the N protein, but two serum samples did not a.a. 180–374 of the N protein. This different reactivity was also confirmed by immunoblot analysis using the serum from the ferret.Therefore, the a.a. 1–179 of the N protein was used as an ELISA antigen. Serological test was carried out using sera or plasma of ferrets in Japan. Surprisingly, 89% ferrets in Japan had been infected with FRCoV. These results indicated that our established ELISA using a.a. 1–179 of the N protein is useful for detection of antibody to FRCoV for diagnosis and seroepidemiology of FRCoV infection. url: https://doi.org/10.1292/jvms.16-0059 doi: 10.1292/jvms.16-0059 id: cord-265312-yfjme53q author: Magtoto, Ronaldo title: Evaluation of the Serologic Cross-Reactivity between Transmissible Gastroenteritis Coronavirus and Porcine Respiratory Coronavirus Using Commercial Blocking Enzyme-Linked Immunosorbent Assay Kits date: 2019-03-13 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: This study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples (n = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue (n = 12), TGEV strain Miller (n = 12), PRCV (n = 12), or with virus-free culture medium (n = 12). ELISA results were evaluated both with “suspect” results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (P < 0.05) cross-reactivity between TGEV and PRCV serum antibodies, particularly during early stages of the infections. Serologic cross-reactivity between TGEV and PRCV seemed to be TGEV strain dependent, with a higher percentage of PRCV-false-positive results for pigs inoculated with TGEV Purdue than for TGEV Miller. Moreover, the overall proportion of false positives was higher when suspect results were interpreted as positive, regardless of the ELISA kit evaluated. IMPORTANCE Current measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals. However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis. url: https://doi.org/10.1128/msphere.00017-19 doi: 10.1128/msphere.00017-19 id: cord-277265-p8pns7r9 author: Malik, Yashpal Singh title: Biotechnological innovations in farm and pet animal disease diagnosis date: 2019-09-20 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: The application of innovative diagnostic technologies for the detection of animal pathogens at an early stage is essential in restricting the economic loss incurred due to emerging infectious animal diseases. The desirable characteristics of such diagnostic methods are easy to use, cost-effective, highly sensitive, and specific, coupled with the high-throughput detection capabilities. The enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) are still the most common assays used for the detection of animal pathogens across the globe. However, utilizing the principles of ELISA and PCR, several serological and molecular technologies have been developed to achieve higher sensitivity, rapid, and point-of-care (POC) detection such as lateral flow assays, biosensors, loop-mediated isothermal amplification, recombinase polymerase amplification, and molecular platforms for field-level detection of animal pathogens. Furthermore, animal disease diagnostics need to be updated regularly to capture new, emerging and divergent infectious pathogens, and biotechnological innovations are helpful in fulfilling the rising demand for such diagnostics for the welfare of the society. Therefore, this chapter primarily describes and discusses in detail the serological, molecular, novel high-throughput, and POC assays to detect pathogens affecting farm and companion animals. url: https://www.sciencedirect.com/science/article/pii/B9780128163528000138 doi: 10.1016/b978-0-12-816352-8.00013-8 id: cord-311349-145kwny3 author: Mariani, Stefano title: Surface plasmon resonance applications in clinical analysis date: 2014-02-25 words: 13425.0 sentences: 630.0 pages: flesch: 39.0 cache: ./cache/cord-311349-145kwny3.txt txt: ./txt/cord-311349-145kwny3.txt summary: In the last 20 years, surface plasmon resonance (SPR) and its advancement with imaging (SPRi) emerged as a suitable and reliable platform in clinical analysis for label-free, sensitive, and real-time monitoring of biomolecular interactions. The advantages brought about by current SPR technology include real-time monitoring of the analyte/molecular markers, label free and parallel analysis (with SPRi), minimal sample pretreatment, quantitative response, and very good sensitivity and reproducibility, (reported detection limits are in atto-or femtomolar ranges and coefficient of variations below 10 %). Preventing nonspecific adsorption of biomolecules (e.g., protein) on the SPR sensing surface is another key-step for the development of specific biosensor, with real application to clinical diagnostics where complex matrices (such as serum, blood, and urine) are analyzed. abstract: In the last 20 years, surface plasmon resonance (SPR) and its advancement with imaging (SPRi) emerged as a suitable and reliable platform in clinical analysis for label-free, sensitive, and real-time monitoring of biomolecular interactions. Thus, we report in this review the state of the art of clinical target detection with SPR-based biosensors in complex matrices (e.g., serum, saliva, blood, and urine) as well as in standard solution when innovative approaches or advanced instrumentations were employed for improved detection. The principles of SPR-based biosensors are summarized first, focusing on the physical properties of the transducer, on the assays design, on the immobilization chemistry, and on new trends for implementing system analytical performances (e.g., coupling with nanoparticles (NPs). Then we critically review the detection of analytes of interest in molecular diagnostics, such as hormones (relevant also for anti-doping control) and biomarkers of interest in inflammatory, cancer, and heart failure diseases. Antibody detection is reported in relation to immune disorder diagnostics. Subsequently, nucleic acid targets are considered for revealing genetic diseases (e.g., point mutation and single nucleotides polymorphism, SNPs) as well as new emerging clinical markers (microRNA) and for pathogen detection. Finally, examples of pathogen detection by immunosensing were also analyzed. A parallel comparison with the reference methods was duly made, indicating the progress brought about by SPR technologies in clinical routine analysis. url: https://www.ncbi.nlm.nih.gov/pubmed/24566759/ doi: 10.1007/s00216-014-7647-5 id: cord-351498-bmq6zcb0 author: Martínez-Sernández, Victoria title: Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis date: 2018-03-21 words: 8277.0 sentences: 401.0 pages: flesch: 48.0 cache: ./cache/cord-351498-bmq6zcb0.txt txt: ./txt/cord-351498-bmq6zcb0.txt summary: In the present study, we developed and tested three indirect ELISAs using rFhpCL1, rFhpCL2, and rFhpCL5 and evaluated their recognition by sera from sheep and cattle naturally infected with F. In the present study, we developed and tested indirect ELISAs based on recombinant procathepsin L1 (rFhpCL1), rFhpCL2, or rFhpCL5, in order to investigate which of these targets are best recognized by sera from sheep and cattle naturally infected with F. Finally, the r values obtained on comparing the four ELISA methods for Fasciola-infected cattle and sheep sera are shown in Table 3 . These results suggest that the use of a single recombinant cathepsin/ procathepsin as target antigen in ELISAs for serodiagnosis of fascioliasis may limit the sensitivity of the assay when testing sera from some species, particularly cattle. (2001) who tested sera from sheep and cattle harboring other parasites, mainly nematodes, suggesting that the cross-reactivity may be due to common epitopes between recombinant Fasciola cathepsins and antigens present in other parasites. abstract: Infections caused by Fasciola hepatica are of great importance in the veterinary field, as they cause important economic losses to livestock producers. Serodiagnostic methods, typically ELISA (with either native or recombinant antigens), are often used for early diagnosis. The use of native antigens, as in the MM3-SERO ELISA (commercialized as BIO K 211, BIO-X Diagnostics), continues to be beneficial in terms of sensitivity and specificity; however, there is interest in developing ELISA tests based on recombinant antigens to avoid the need to culture parasites. Of the antigens secreted by adult flukes, recombinant procathepsin L1 (rFhpCL1) is the most commonly tested in ELISA to date. However, although adult flukes produce three different clades of CLs (FhCL1, FhCL2, and FhCL5), to our knowledge, the diagnostic value of recombinant FhCL2 and FhCL5 has not yet been investigated. In the present study, we developed and tested three indirect ELISAs using rFhpCL1, rFhpCL2, and rFhpCL5 and evaluated their recognition by sera from sheep and cattle naturally infected with F. hepatica. Although the overall antibody response to these three rFhpCLs was similar, some animals displayed preferential recognition for particular rFhpCLs. Moreover, for cattle sera, the highest sensitivity was obtained using rFhpCL2 (97%), being equal for both rFhpCL1 and rFhpCL5 (87.9%), after adjusting cut-offs for maximum specificity. By contrast, for sheep sera, the sensitivity was 100% for the three rFhpCLs. Finally, the presence of truncated and/or partially unfolded molecules in antigen preparations is postulated as a possible source of cross-reactivity. url: https://www.ncbi.nlm.nih.gov/pubmed/29564626/ doi: 10.1007/s00436-018-5809-7 id: cord-010578-uib9h1lb author: Mawle, Alison C. title: Seroepidemiology of Chronic Fatigue Syndrome: A Case-Control Study date: 1995-12-17 words: 2570.0 sentences: 164.0 pages: flesch: 49.0 cache: ./cache/cord-010578-uib9h1lb.txt txt: ./txt/cord-010578-uib9h1lb.txt summary: We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We conducted serological tests for a large number of infectious agents as part of a case-control study assessing risk factors for CFS. Antibodies against human T-lymphotrophic virus types I and II were detected with an ELISA, and confirmatory testing was performed by western blotting [5] . All other agents tested were detected in ;;:::25% of CFS cases, and antibody levels were compared between cases and controls. Evidence for active Epstein-Barr virus infection in patients with persistent unexplained illness: elevated anti-early antigen antibodies abstract: We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We did not find any agent associated with CFS. In addition, we did not find elevated levels of antibody to any of a wide range of agents examined. In particular, we did not find elevated titers of antibody to any herpesvirus, nor did we find evidence of enteroviral exposure in this group of patients. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7197952/ doi: 10.1093/clinids/21.6.1386 id: cord-264936-3posyr5n author: Mohammadzadeh, Sara title: Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications date: 2014-11-24 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Hepatitis C virus (HCV) is major cause of liver cirrhosis in humans. HCV capsid (core) protein (HCVcp) is a highly demanded antigen for various diagnostic, immunization and pathogenesis studies. Plants are considered as an expression system for producing safe and inexpensive biopharmaceutical proteins. Although invention of transgenic (stable) tobacco plants expressing HCVcp with proper antigenic properties was recently reported, no data for “transient-expression” that is currently the method of choice for rapid, simple and lower-priced protein expression in plants is available for HCVcp. OBJECTIVES: The purpose of this study was to design a highly codon-optimized HCVcp gene for construction of an efficient transient-plant expression system for production of HCVcp with proper antigenic properties in a regional tobacco plant (Iranian Jafarabadi-cultivar) by evaluation of different classes of vectors and suppression of gene-silencing in tobacco. MATERIALS AND METHODS: A codon-optimized gene encoding the Kozak sequence, 6xHis-tag, HCVcp (1-122) and KDEL peptide in tandem (from N- to C-terminal) was designed and inserted into potato virus-X (PVX) and classic pBI121 binary vectors in separate cloning reactions. The resulted recombinant plasmids were transferred into Agrobacterium tumefaciens and vacuum infiltrated into tobacco leaves. The effect of gene silencing suppressor P19 protein derived from tomato bushy stunt virus on the expression yield of HCVcp by each construct was also evaluated by co-infiltration in separate groups. The expressed HCVcp was evaluated by dot and western blotting and ELISA assays. RESULTS: The codon-optimized gene had an increased adaptation index value (from 0.65 to 0.85) and reduced GC content (from 62.62 to 51.05) in tobacco and removed the possible deleterious effect of “GGTAAG” splice site in native HCVcp. Blotting assays via specific antibodies confirmed the expression of the 15 kDa HCVcp. The expression level of HCVcp was enhanced by 4-5 times in P19 co-agroinfiltrated plants with better outcomes for PVX, compared to pBI121 vector (0.022% versus 0.019% of the total soluble protein). The plant-derived HCVcp (pHCVcp) could properly identify the HCVcp antibody in HCV-infected human sera compared to Escherichia coli-derived HCVcp (eHCVcp), indicating its potential for diagnostic/immunization applications. CONCLUSIONS: By employment of gene optimization strategies, use of viral-based vectors and suppression of plant-derived gene silencing effect, efficient transient expression of HCVcp in tobacco with proper antigenic properties could be possible. url: https://doi.org/10.5812/hepatmon.20524 doi: 10.5812/hepatmon.20524 id: cord-288202-r3r2bc7v author: Morel, Noelia title: A Monoclonal Antibody-Based Copro-ELISA Kit for Canine Echinococcosis to Support the PAHO Effort for Hydatid Disease Control in South America date: 2013-01-10 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Cystic echinococcosis is still a major concern in South America. While some regions show advances in the control of the disease, others have among the highest incidence in the world. To reverse this situation the Pan American Health Organization (PAHO) has launched a regional project on cystic echinococcosis control and surveillance. An early concern of the program was the lack of a standardized diagnostic tool to monitor infection in dogs, a key target of control programs. Under this premise, we have developed a new copro-ELISA test after extensive screening of a large panel of monoclonal antibodies (MAbs) and polyclonal sera, which performs with high standards of sensitivity (92.6%) and specificity (86.4%) as established by necropsy diagnosis of dogs. The key component of the test, MAbEg9 has a convenient IgG isotype and reacts with a periodate-resistant epitope found in high molecular weight components of the worm. Time-course analysis of experimentally infected dogs showed that even animals with a very low number of parasites could be detected as early as day 20 post infection. The test was formulated in a ready-to-use kit format with proven stability of each component for a minimum of 3 months at room temperature. This characteristic facilitates its standardized use and shipping to other laboratories, which was demonstrated by the identical results obtained by two different laboratories in Peru and our own laboratory when a large number of field samples were analyzed independently in a blind fashion. url: https://www.ncbi.nlm.nih.gov/pubmed/23326610/ doi: 10.1371/journal.pntd.0001967 id: cord-298766-xqkre25z author: Muller, Janine D. title: Improvement of a recombinant antibody-based serological assay for foot-and-mouth disease virus date: 2010-01-31 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Abstract Differentiating foot-and-mouth disease virus (FMDV) antibodies generated during a natural infection from those due to vaccination (DIVA) is crucial for proving freedom from disease after an outbreak and allowing resumption of trade in livestock products. The World Organisation for Animal Health (OIE) recommends that FMDV vaccines are composed of inactivated virus that has been purified to remove non-structural viral proteins. Such purified vaccines primarily induce antibodies to viral structural proteins, whereas replicating virus stimulates host antibodies specific for both structural and non-structural proteins. The current preferred FMDV DIVA test is a competitive ELISA (C-ELISA) designed to detect antibodies to the non-structural protein 3ABC. Previously, we described the development of an FMDV DIVA test based entirely on recombinant proteins (the recombinant detecting antibody and the 3ABC coating antigen) produced in Escherichia coli. In this study, we have determined the precise binding site of the recombinant detecting antibody to a conserved sequence within the 3B region of the 3ABC protein, replaced the original E-tag of the detecting antibody with two in-house tags and engineered a direct antibody–reporting enzyme (alkaline phosphatase) fusion protein. These modifications have further improved the DIVA test, providing great potential for large scale production and uptake due to its simplicity, reproducibility and low cost. url: https://doi.org/10.1016/j.jim.2009.11.004 doi: 10.1016/j.jim.2009.11.004 id: cord-262640-4vr4cm1s author: Nguyen, N. N. title: Correlation of ELISA based with random access serologic immunoassays for identifying adaptive immune response to SARS-CoV-2 date: 2020-07-08 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Public health emergency of SARS-CoV-2 has facilitated diagnostic testing as a related medical countermeasure against COVID-19 outbreak. Numerous serologic antibody tests have become available through an expedited federal emergency use only process. This paper highlights the analytical characteristic of an ELISA based assay by AnshLabs and three random access immunoassay (RAIA) by DiaSorin, Roche, and Abbott that have been approved for emergency use authorization (EUA), at a tertiary academic center in a low disease-prevalence area. The AnshLabs gave higher estimates of sero-prevalence, over the three RAIA methods. For positive results, AnshLabs had 93.3% and 100% concordance with DiaSorin or Abbott and Roche respectively. For negative results, AnshLabs had 69.7% and 73.0% concordance with DiaSorin and Roche or Abbott respectively. All discrepant samples that were positive by AnshLabs and negative by RAIA tested positive by all-in-one step SARS-CoV-2 Total (COV2T) assay performed on the automated Siemens Advia Centaur XPT analyzer. None of these methods, however, are useful in early diagnosis of SARSCoV- 2. url: https://doi.org/10.1101/2020.07.06.20145938 doi: 10.1101/2020.07.06.20145938 id: cord-309415-77b5erfj author: Nguyen, T. D. title: Étude comparée de trois souches du coronavirus de la gastroentérite transmissible: Conditions de la réplicationvirale et de la synthèse des antigènes structuraux date: 1987-09-30 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Summary Purdue-115 and D-52 strains of TGEV were compared with the 188-SG strain, which was obtained by means of a survivor selection process in gastric juice of adult pig. The 188-SG strain was characterized by (a) low infectivity, (b) delayed and restricted growth associated with low and delayed RNA synthesis, and (c) a high content of structural antigens. In contrast, Purdue-115 and D-52 strains were characterized by (a) high infectivity, and (b) a normal pattern of virus replication and RNA and structural antigen synthesis. Tunicamycin induced the inhibition of synthesis of El and E2 glycoproteins (detected by the ELISA test using monoclonal and polyclonal antibodies) as well as a significant reduction in the NP protein. The inhibitory effect of tunicamycin was influenced by the cell type and virus strain. url: https://www.sciencedirect.com/science/article/pii/S0769261787800187 doi: 10.1016/s0769-2617(87)80018-7 id: cord-353190-7qcoxl81 author: Nicklas, Werner title: Viral Infections of Laboratory Mice date: 2012-05-17 words: 27775.0 sentences: 1482.0 pages: flesch: 39.0 cache: ./cache/cord-353190-7qcoxl81.txt txt: ./txt/cord-353190-7qcoxl81.txt summary: This chapter covers infections of mice with the following viruses: herpesviruses, mousepox virus, murine adenoviruses, polyomaviruses, parvoviruses, lactate dehydrogenase-elevating virus, lymphocytic choriomeningitis virus, mammalian orthoreovirus serotype 3, murine hepatitis virus, murine norovirus, murine pneumonia virus, murine rotavirus, Sendai virus, and Theiler''s murine encephalomyelitis virus. These results are very difficult to summarize because the outcome of experimental infection in laboratory mice depends on various factors such as mouse strain and age, virus strain and passage history [26] , virus dose and route of inoculation [24] . Experimental infection of laboratory mice with MHV-68 is a frequently used model system for the study of human gammaherpesvirus pathogenesis, e.g. of Kaposi''s sarcoma-associated herpesvirus or Epstein-Barr virus (EBV) [62, 63] which are members of the same subfamily. Early descriptions of naturally occurring disease may have been complicated by concurrent infections such as MHV (murine hepatitis virus) or murine rotavirus A (MuRV-A)/epizootic diarrhoea of infant mice (EDIM) virus that contributed to the severity of the lesions especially in liver, pancreas, CNS and intestine. abstract: Viral infections of laboratory mice have considerable impact on research results, and prevention of such infections is therefore of crucial importance. This chapter covers infections of mice with the following viruses: herpesviruses, mousepox virus, murine adenoviruses, polyomaviruses, parvoviruses, lactate dehydrogenase-elevating virus, lymphocytic choriomeningitis virus, mammalian orthoreovirus serotype 3, murine hepatitis virus, murine norovirus, murine pneumonia virus, murine rotavirus, Sendai virus, and Theiler’s murine encephalomyelitis virus. For each virus, there is a description of the agent, epizootiology, clinical symptoms, pathology, methods of diagnosis and control, and its impact on research. url: https://api.elsevier.com/content/article/pii/B9780123820082000192 doi: 10.1016/b978-0-12-382008-2.00019-2 id: cord-284841-flhfagp3 author: Nicol, Thomas title: Assessment of SARS-CoV-2 serological tests for the diagnosis of COVID-19 through the evaluation of three immunoassays: two automated immunoassays (Euroimmun and Abbott) and one rapid lateral flow immunoassay (NG Biotech) date: 2020-06-15 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: The emergence of new SARS-CoV-2 has promoted the development of new serological tests that could be complementary to RT-PCR. Nevertheless, the assessment of clinical performances of available tests is urgently required as their use has just been initiated for diagnose. OBJECTIVES: The aim of this study was to assess the performance of three immunoassays for the detection of SARS-CoV-2 antibodies. METHODS: Two automated immunoassays (Abbott SARS-CoV-2 CLIA IgG and Euroimmun Anti-SARS-CoV-2 ELISA IgG/IgA assays) and one lateral flow immunoassay (LFIA NG-Test® IgG-IgM COVID-19) were tested. 293 specimens were analyzed from patients with a positive RT-PCR response, from patients with symptoms consistent with COVID-19 but exhibiting a negative response to the RT-PCR detection test, and from control group specimens. Days since symptoms onset were collected from clinical information sheet associated with respiratory tract samples. RESULTS: Overall sensitivity for IgG was equivalent (around 80%) for CLIA, ELISA and LFIA. Sensitivity for IgG detection, >14 days after onset of symptoms, was 100.0% for all assays. Overall specificity for IgG was greater for CLIA and LFIA (more than 98%) compared to ELISA (95.8%). Specificity was significantly different between IgA ELISA (78.9%) and IgM LFIA (95.8%) (p < 0.05). The best agreement was observed between CLIA and LFIA assays (97%; k = 0.936). CONCLUSION: Excellent sensitivity for IgG detection was obtained >14 days after onset of symptoms for all immunoassays. Specificity was also excellent for IgG CLIA and IgG LFIA. Our study shows that NG-Test® is reliable and accurate for routine use in clinical laboratories. url: https://doi.org/10.1016/j.jcv.2020.104511 doi: 10.1016/j.jcv.2020.104511 id: cord-301313-9595vm0k author: OKBA, NISREEN M.A. title: SARS-CoV-2 specific antibody responses in COVID-19 patients date: 2020-03-20 words: 4271.0 sentences: 248.0 pages: flesch: 55.0 cache: ./cache/cord-301313-9595vm0k.txt txt: ./txt/cord-301313-9595vm0k.txt summary: Here, we describe development of serological assays for the detection of virus neutralizing antibodies and antibodies to the nucleocapsid (N) protein and various spike (S) domains including the S1 subunit, and receptor binding domain (RBD) of SARS-CoV-2 in ELISA format. Using a wellcharacterized cohort of serum samples from PCR-confirmed SARS-CoV-2 and patients PCR-confirmed to be infected with seasonal coronaviruses and other respiratory pathogens, we validated and tested various antigens in different platforms developed in-house as well as a commercial platform. We evaluated SARS-CoV-2 specific antibody responses in severe and mild cases using serum samples collected at different times post-disease onset from three French PCR-confirmed CoVID-19 patients. We tested sera for SARS-CoV-2 specific antibodies using different ELISAs. Following infections, all three patients seroconverted between days 13 and 21 post onset of disease (Figure 1) , and antibodies were elicited against the SARS-CoV-2 S and S1 subunit including the N-terminal (S1 A ) domain and the receptor binding domain (RBD). abstract: A new coronavirus, SARS-CoV-2, has recently emerged to cause a human pandemic. Whereas molecular diagnostic tests were rapidly developed, serologic assays are still lacking, yet urgently needed. Validated serologic assays are important for contact tracing, identifying the viral reservoir and epidemiological studies. Here, we developed serological assays for the detection of SARS-CoV-2 neutralizing, spike- and nucleocapsid-specific antibodies. Using serum samples from patients with PCR-confirmed infections of SARS-CoV-2, other coronaviruses, or other respiratory pathogenic infections, we validated and tested various antigens in different in-house and commercial ELISAs. We demonstrate that most PCR-confirmed SARS-CoV-2 infected individuals seroconverted, as revealed by sensitive and specific in-house ELISAs. We found that commercial S1 IgG or IgA ELISAs were of lower specificity while sensitivity varied between the two, with IgA showing higher sensitivity. Overall, the validated assays described here can be instrumental for the detection of SARS-CoV-2-specific antibodies for diagnostic, seroepidemiological and vaccine evaluation studies. url: https://doi.org/10.1101/2020.03.18.20038059 doi: 10.1101/2020.03.18.20038059 id: cord-335591-r0x8yaqj author: Ohnishi, Kazuo title: Establishment and Characterization of Monoclonal Antibodies Against SARS Coronavirus date: 2007-11-28 words: 3126.0 sentences: 242.0 pages: flesch: 67.0 cache: ./cache/cord-335591-r0x8yaqj.txt txt: ./txt/cord-335591-r0x8yaqj.txt summary: The hybridomas produce monoclonal antibodies that recognize viral component molecules, including the spike protein (S) and the nucleocapsid protein (N), enabling the immunological detection of SARS-CoV by immunofluorescence staining, immunoblot, or an antigen-capture ELISA system. Based on clinical experience, several options have been considered in the quest to develop the capacity to accurately diagnose SARS-CoV infection, including molecular biology techniques and serological tests such as antigen-capture ELISA assay and immunofluorescence assay to detect virus-infected cells in respiratory swabs (3-7) . These mAbs enable the general immunological detection of SARS-CoV by methods such as immunofluorescent staining, immunoblotting, and immunohistology, in addition to the construction of a highly sensitive antigen-capture sandwich ELISA (6). The UV-inactivated purified SARS-CoV samples (see Note 1), which are serially diluted with 1% OVA/PBS-Tween, are added to the wells and incubated for 1 h at room temperature abstract: Immunological detection of viruses and their components by monoclonal antibodies is a powerful method for studying the structure and function of viral molecules. Here we describe detailed methods for establishing monoclonal antibodies against severe acute respiratory syndrome coronavirus (SARS-CoV). B cell hybridomas are generated from mice that are hyperimmunized with inactivated SARS-CoV virions. The hybridomas produce monoclonal antibodies that recognize viral component molecules, including the spike protein (S) and the nucleocapsid protein (N), enabling the immunological detection of SARS-CoV by immunofluorescence staining, immunoblot, or an antigen-capture ELISA system. In addition, several S protein-specific antibodies are shown to have in vitro neutralization activity. Thus the monoclonal antibody approach provides useful tools for rapid and specific diagnosis of SARS, as well as for possible antibody-based treatment of the disease. url: https://doi.org/10.1007/978-1-59745-181-9_15 doi: 10.1007/978-1-59745-181-9_15 id: cord-347374-mryazbnq author: Okba, Nisreen M.A. title: Severe Acute Respiratory Syndrome Coronavirus 2−Specific Antibody Responses in Coronavirus Disease Patients date: 2020-07-17 words: 3565.0 sentences: 186.0 pages: flesch: 51.0 cache: ./cache/cord-347374-mryazbnq.txt txt: ./txt/cord-347374-mryazbnq.txt summary: Using serum samples from patients with PCR-confirmed SARS-CoV-2 infections, other coronaviruses, or other respiratory pathogenic infections, we validated and tested various antigens in different in-house and commercial ELISAs. We demonstrated that most PCR-confirmed SARS-CoV-2–infected persons seroconverted by 2 weeks after disease onset. Using a well-characterized cohort of serum samples from PCR-confirmed SARS-CoV-2 and patients PCR-confirmed to be infected with seasonal coronaviruses and other respiratory pathogens, we validated and tested various antigens in different platforms developed in-house, as well as a commercial platform. We evaluated SARS-CoV-2-specific antibody responses in severe and mild cases by using serum samples collected at different times postonset of disease from 3 PCR-confirmed COVID-19 patients from France. We tested serum samples for SARS-CoV-2specific antibodies by using different ELISAs. After infection, all 3 patients seroconverted between days 13 and 21 after onset of disease (Figure 1) , and antibodies were elicited against the SARS-CoV-2 S, S1 subunit, and RBD, but only 2/3 patients had detectable antibodies to the N-terminal (S1 A ) domain. abstract: A new coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has recently emerged to cause a human pandemic. Although molecular diagnostic tests were rapidly developed, serologic assays are still lacking, yet urgently needed. Validated serologic assays are needed for contact tracing, identifying the viral reservoir, and epidemiologic studies. We developed serologic assays for detection of SARS-CoV-2 neutralizing, spike protein–specific, and nucleocapsid-specific antibodies. Using serum samples from patients with PCR-confirmed SARS-CoV-2 infections, other coronaviruses, or other respiratory pathogenic infections, we validated and tested various antigens in different in-house and commercial ELISAs. We demonstrated that most PCR-confirmed SARS-CoV-2–infected persons seroconverted by 2 weeks after disease onset. We found that commercial S1 IgG or IgA ELISAs were of lower specificity, and sensitivity varied between the 2 assays; the IgA ELISA showed higher sensitivity. Overall, the validated assays described can be instrumental for detection of SARS-CoV-2–specific antibodies for diagnostic, seroepidemiologic, and vaccine evaluation studies. url: https://www.ncbi.nlm.nih.gov/pubmed/32267220/ doi: 10.3201/eid2607.200841 id: cord-301175-6alsigxk author: Okda, Faten title: Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus date: 2015-08-01 words: 8275.0 sentences: 411.0 pages: flesch: 46.0 cache: ./cache/cord-301175-6alsigxk.txt txt: ./txt/cord-301175-6alsigxk.txt summary: title: Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of Porcine Epidemic Diarrhea Virus Therefore, the objective of this study was to develop and validate multiple improved serological assays for PEDV, including an indirect ELISA (iELISA); a highly specific monoclonal antibody-based blocking ELISA (bELISA); fluorescent microsphere immunoassays (FMIA) that can be multiplexed to monitor exposure to multiple antigens and pathogens simultaneously; and a fluorescent focus neutralization assay (FFN) to measure functional virus neutralizing antibodies. RESULTS: A recombinant North American nucleoprotein (NP) based iELISA was developed and validated along with a bELISA using newly developed PEDV-NP specific biotinylated monoclonal antibodies (mAbs) and an FMIA using magnetic beads coupled with expressed NA PEDV-NP. In this study, we report the adaptation of a recombinant, highly purified, NA PEDV-NP antigen to the development of iELISA, bELISA and FMIA platforms for the detection of PEDV antibodies in serum. abstract: BACKGROUND: Recent, severe outbreaks of porcine epidemic diarrhea virus (PEDV) in Asia and North America highlight the need for well-validated diagnostic tests for the identification of PEDV infected animals and evaluation of their immune status to this virus. PEDV was first detected in the U.S. in May 2013 and spread rapidly across the country. Some serological assays for PEDV have been previously described, but few were readily available in the U.S. Several U.S. laboratories quickly developed indirect fluorescent antibody (IFA) assays for the detection of antibodies to PEDV in swine serum, indicating prior exposure. However, the IFA has several disadvantages, including low throughput and relatively subjective interpretation. Different serologic test formats have advantages and disadvantages, depending on the questions being asked, so a full repertoire of tests is useful. Therefore, the objective of this study was to develop and validate multiple improved serological assays for PEDV, including an indirect ELISA (iELISA); a highly specific monoclonal antibody-based blocking ELISA (bELISA); fluorescent microsphere immunoassays (FMIA) that can be multiplexed to monitor exposure to multiple antigens and pathogens simultaneously; and a fluorescent focus neutralization assay (FFN) to measure functional virus neutralizing antibodies. RESULTS: A recombinant North American nucleoprotein (NP) based iELISA was developed and validated along with a bELISA using newly developed PEDV-NP specific biotinylated monoclonal antibodies (mAbs) and an FMIA using magnetic beads coupled with expressed NA PEDV-NP. Receiver operating characteristic (ROC) analysis was performed using swine serum samples (iELISA n = 1486, bELISA n = 1186, FMIA n = 1420). The ROC analysis for the FMIA showed estimated sensitivity and specificity of 98.2 and 99.2 %, respectively. The iELISA and bELISA showed a sensitivity and specificity of 97.9 and 97.6 %; and 98.2 and 98.9 %, respectively. Inter-rater (kappa) agreement was calculated to be 0.941 between iELISA and IFA, 0.945 between bELISA and IFA and 0.932 between FMIA and IFA. Similar comparative kappa values were observed between the iELISA, bELISA and FMIA, which demonstrated a significant level of testing agreement among the three assays. No cross-reactivity with the closely related coronaviruses, transmissible gastroenteritis virus (TGEV) or porcine respiratory coronavirus (PRCV) was noted with these assays. All three assays detected seroconversion of naïve animals within 6–9 days post exposure. The FFN assay allows relative quantitation of functional neutralizing antibodies in serum, milk or colostrum samples. CONCLUSION: Well-validated iELISA, bELISA and FMIA assays for the detection of PEDV antibodies were developed and showed good correlation with IFA and each other. Each assay format has advantages that dictate how they will be used in the field. Newly developed mAbs to the PEDV-NP were used in the bELISA and for expediting FFN testing in the detection and quantitation of neutralizing antibodies. In addition, these PEDV mAbs are useful for immunohistochemistry, fluorescent antibody staining and other antigen-based tests. Measurement of neutralizing antibody responses using the FFN assay may provide a valuable tool for assessment of vaccine candidates or protective immunity. url: https://doi.org/10.1186/s12917-015-0500-z doi: 10.1186/s12917-015-0500-z id: cord-344581-h7ikjgic author: Ong, David S.Y. title: Comparison of diagnostic accuracies of rapid serological tests and ELISA to molecular diagnostics in patients with suspected COVID-19 presenting to the hospital date: 2020-06-02 words: 2071.0 sentences: 120.0 pages: flesch: 54.0 cache: ./cache/cord-344581-h7ikjgic.txt txt: ./txt/cord-344581-h7ikjgic.txt summary: OBJECTIVES: To assess the diagnostic performance of rapid lateral flow immunochromatographic assays (LFAs) compared to an enzyme-linked immunosorbent assay (ELISA) and nucleic acid amplification tests (NATs) in suspected coronavirus disease 2019 (COVID-19) patients. In the total cohort, Orient Gene Biotech COVID-19 IgG/IgM Rapid Test LFA had a sensitivity of 43/99 (43%; 95% CI 34-53) and specificity of 126/129 (98%; 95% CI 95-100). CONCLUSIONS: There is large variability in diagnostic test performance between rapid LFAs, but overall limited sensitivity and high specificity in acutely admitted patients. First, in a pilot phase 20 NAT-positive and 5 NAT-negative patients were retrospectively selected for which six LFAs were performed on heparin plasma samples obtained upon hospital presentation ( Figure S1 ), which corresponded to the dates of molecular testing. This study shows that the sensitivity of LFA was low in patients suspected for COVID-19 presenting to the hospital, but it improved in patients with at least seven days of symptoms and in those with CRP levels >100 mg/L upon presentation. abstract: OBJECTIVES: To assess the diagnostic performance of rapid lateral flow immunochromatographic assays (LFAs) compared to an enzyme-linked immunosorbent assay (ELISA) and nucleic acid amplification tests (NATs) in suspected coronavirus disease 2019 (COVID-19) patients. METHODS: Patients presenting to a Dutch teaching hospital were eligible between March 17 and April 10, 2020, when they had respiratory symptoms that were suspected for COVID-19. The performances of six different LFAs were evaluated in plasma samples obtained on corresponding respiratory sample dates of NATs testing. Subsequently, the best performing LFA was evaluated in 228 patients and in 50 sera of a historical patient control group. RESULTS: In the pilot analysis sensitivity characteristics of LFA were heterogenous ranging from 2/20 (10%; 95% confidence interval (CI) 0-23) to 11/20 (55%; 95% CI 33-77). In the total cohort, Orient Gene Biotech COVID-19 IgG/IgM Rapid Test LFA had a sensitivity of 43/99 (43%; 95% CI 34-53) and specificity of 126/129 (98%; 95% CI 95-100). Sensitivity increased to 31/52 (60%; 95% CI 46-73) in patients with at least seven days of symptoms, and to 21/33 (64%; 95% CI 47-80) in patients with C-reactive protein (CRP) >100 mg/L. Sensitivity and specificity of Wantai SARS-CoV-2 Ab ELISA was 59/95 (62%; 95% CI 52-72) and 125/128 (98%; 95% CI 95-100) in all patients, respectively, but sensitivity increased to 38/48 (79%; 95% CI 68-91) in patients with at least seven days of symptoms. CONCLUSIONS: There is large variability in diagnostic test performance between rapid LFAs, but overall limited sensitivity and high specificity in acutely admitted patients. Sensitivity improved in patients with longer existing symptoms or high CRP. LFAs should only be considered as additional triage tools when these may lead to the improvement of hospital logistics. url: https://doi.org/10.1016/j.cmi.2020.05.028 doi: 10.1016/j.cmi.2020.05.028 id: cord-327890-ocisq7e4 author: Pallett, Scott J C title: Point-of-care serological assays for delayed SARS-CoV-2 case identification among health-care workers in the UK: a prospective multicentre cohort study date: 2020-07-24 words: 5947.0 sentences: 271.0 pages: flesch: 42.0 cache: ./cache/cord-327890-ocisq7e4.txt txt: ./txt/cord-327890-ocisq7e4.txt summary: In phase 1, two point-of-care lateral flow serological assays, the Onsite CTK Biotech COVID-19 split IgG/IgM Rapid Test (CTK Bitotech, Poway, CA, USA) and the Encode SARS-CoV-2 split IgM/IgG One Step Rapid Test Device (Zhuhai Encode Medical Engineering, Zhuhai, China), were evaluated for performance against a laboratory immunoassay (EDI Novel Coronavirus COVID-19 IgG ELISA kit [Epitope Diagnostics, San Diego, CA, USA]) in 300 samples from health-care workers and 100 pre-COVID-19 negative control samples. These include point-of-care molecular platforms for acute phase testing, and laboratory ELISA or lateral flow serological assays for antibodies specific to SARS-CoV-2 for delayed case identification. To derive a measure of sensitivity, the results of lateral flow serological assays and ELISA were compared in 300 health-care workers who had previously received PCR testing (AusDiagnostics, Sydney, Australia) at initial presentation with COVID-19 symptoms. abstract: BACKGROUND: Health-care workers constitute a high-risk population for acquisition of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Capacity for acute diagnosis via PCR testing was limited for individuals with mild to moderate SARS-CoV-2 infection in the early phase of the COVID-19 pandemic and a substantial proportion of health-care workers with suspected infection were not tested. We aimed to investigate the performance of point-of-care and laboratory serology assays and their utility in late case identification, and to estimate SARS-CoV-2 seroprevalence. METHODS: We did a prospective multicentre cohort study between April 8 and June 12, 2020, in two phases. Symptomatic health-care workers with mild to moderate symptoms were eligible to participate 14 days after onset of COVID-19 symptoms, as per the Public Health England (PHE) case definition. Health-care workers were recruited to the asymptomatic cohort if they had not developed PHE-defined COVID-19 symptoms since Dec 1, 2019. In phase 1, two point-of-care lateral flow serological assays, the Onsite CTK Biotech COVID-19 split IgG/IgM Rapid Test (CTK Bitotech, Poway, CA, USA) and the Encode SARS-CoV-2 split IgM/IgG One Step Rapid Test Device (Zhuhai Encode Medical Engineering, Zhuhai, China), were evaluated for performance against a laboratory immunoassay (EDI Novel Coronavirus COVID-19 IgG ELISA kit [Epitope Diagnostics, San Diego, CA, USA]) in 300 samples from health-care workers and 100 pre-COVID-19 negative control samples. In phase 2 (n=6440), serosurveillance was done among 1299 (93·4%) of 1391 health-care workers reporting symptoms, and in a subset of asymptomatic health-care workers (405 [8·0%] of 5049). FINDINGS: There was variation in test performance between the lateral flow serological assays; however, the Encode assay displayed reasonable IgG sensitivity (127 of 136; 93·4% [95% CI 87·8–96·9]) and specificity (99 of 100; 99·0% [94·6–100·0]) among PCR-proven cases and good agreement (282 of 300; 94·0% [91·3–96·7]) with the laboratory immunoassay. By contrast, the Onsite assay had reduced sensitivity (120 of 136; 88·2% [95% CI 81·6–93·1]) and specificity (94 of 100; 94·0% [87·4–97·8]) and agreement (254 of 300; 84·7% [80·6–88·7]). Five (7%) of 70 PCR-positive cases were negative across all assays. Late changes in lateral flow serological assay bands were recorded in 74 (9·3%) of 800 cassettes (35 [8·8%] of 400 Encode assays; 39 [9·8%] of 400 Onsite assays), but only seven (all Onsite assays) of these changes were concordant with the laboratory immunoassay. In phase 2, seroprevalence among the workforce was estimated to be 10·6% (95% CI 7·6–13·6) in asymptomatic health-care workers and 44·7% (42·0–47·4) in symptomatic health-care workers. Seroprevalence across the entire workforce was estimated at 18·0% (95% CI 17·0–18·9). INTERPRETATION: Although a good positive predictive value was observed with both lateral flow serological assays and ELISA, this agreement only occurred if the pre-test probability was modified by a strict clinical case definition. Late development of lateral flow serological assay bands would preclude postal strategies and potentially home testing. Identification of false-negative results among health-care workers across all assays suggest caution in interpretation of IgG results at this stage; for now, testing is perhaps best delivered in a clinical setting, supported by government advice about physical distancing. FUNDING: None. url: https://www.ncbi.nlm.nih.gov/pubmed/32717210/ doi: 10.1016/s2213-2600(20)30315-5 id: cord-011212-ovjdzyxv author: Pan, Qing title: Development and application of a novel ELISA for detecting antibodies against group I fowl adenoviruses date: 2019-12-14 words: 3884.0 sentences: 223.0 pages: flesch: 47.0 cache: ./cache/cord-011212-ovjdzyxv.txt txt: ./txt/cord-011212-ovjdzyxv.txt summary: Since 2015, outbreaks of hepatitis-hydropericardium syndrome (HPS) caused by a novel genotype of fowl adenovirus 4 (FAdV-4) infection have created serious economic losses in China. In one recent study, recombinant fiber-based indirect ELISA was used to detect serum samples from chickens experimentally inoculated with different FAdV-1 or FAdV-4 strains (Feichtner et al. A recombinant hexon-based single serum dilution ELISA was also developed to measure the hexon-specific antibodies against FAdV-4 in sera of chickens (Rajasekhar and Roy 2014) . In this study, we developed a group-specific and sensitive ELISA based on the novel genotype of FAdV-4 for detecting antibodies against twelve FAdV-I serotypes. The common ELISA developed in our study was applied to detect serum samples from SPF chickens inoculated with inactivated FAdV-1, FAdV-4, and FAdV-8a, and showed high sensitivity for all three FAdV hypervirulent serotypes. Furthermore, the ELISA showed higher sensitivity in detecting serum samples of HPS caused by the novel FAdV-4 genotype that has recently emerged in China. abstract: Since 2015, outbreaks of hepatitis-hydropericardium syndrome (HPS) caused by a novel genotype of fowl adenovirus 4 (FAdV-4) infection have created serious economic losses in China. Given that other serotypes of hypervirulent FAdVs have also been reported in poultry around the world, a common ELISA for all serotypes within the group I fowl adenoviruses (FAdV-I) is urgently needed, especially for clinical epidemic serotypes. In this study, we used high purity and concentration virions of FAdV-4 and developed a common ELISA for detecting antibodies against 12 FAdV-I serotypes. The developed ELISA was able to distinguish between antibodies against FAdV-I, FAdV-III, and other heterologous viruses without any cross-reaction. Furthermore, the ELISA showed higher sensitivity than the FAdV-1-based ELISA to the novel FAdV-4 found in China. Moreover, since there are no commercial vaccines against FAdVs in China, the ELISA was applied to detect sera samples from specific pathogen-free chickens inoculated with inactivated FAdV-1, FAdV-4, and FAdV-8a. The assay showed high sensitivities for all three detected serotypes within FAdV-I. In conclusion, a novel, common ELISA for FAdV-I was developed in this study and could be a powerful tool for seroepidemiological investigations and FAdVs vaccine development. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7223807/ doi: 10.1007/s00253-019-10208-3 id: cord-284045-scd3f8vk author: Pape, Constantin title: Microscopy-based assay for semi-quantitative detection of SARS-CoV-2 specific antibodies in human sera date: 2020-10-07 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Emergence of the novel pathogenic coronavirus SARS-CoV-2 and its rapid pandemic spread presents numerous questions and challenges that demand immediate attention. Among these is the urgent need for a better understanding of humoral immune response against the virus as a basis for developing public health strategies to control viral spread. For this, sensitive, specific and quantitative serological assays are required. Here we describe the development of a semi-quantitative high-content microscopy-based assay for detection of three major classes (IgG, IgA and IgM) of SARS-CoV-2 specific antibodies in human samples. The possibility to detect antibodies against the entire viral proteome together with a robust semi-automated image analysis workflow resulted in specific, sensitive and unbiased assay which complements the portfolio of SARS-CoV-2 serological assays. The procedure described here has been used for clinical studies and provides a general framework for the application of quantitative high-throughput microscopy to rapidly develop serological assays for emerging virus infections. url: https://doi.org/10.1101/2020.06.15.152587 doi: 10.1101/2020.06.15.152587 id: cord-003859-k8wfyj9b author: Paweska, Janusz T. title: Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera date: 2019-07-24 words: 6407.0 sentences: 308.0 pages: flesch: 51.0 cache: ./cache/cord-003859-k8wfyj9b.txt txt: ./txt/cord-003859-k8wfyj9b.txt summary: title: Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera Diagnostic performance of three indirect enzyme-linked immunosorbent assays (I-ELISA) was evaluated for the detection of IgG antibody to Ebola virus (EBOV) in human sera. Validation data sets derived from individual sera collected in South Africa (SA), representing an EBOV non-endemic country, and from sera collected during an Ebola disease (EBOD) outbreak in Sierra Leone (SL), were categorized according to the compounded results of the three I-ELISAs and real time reverse-transcription polymerase chain reaction (RT-PCR). The purpose of this study was to evaluate and compare the diagnostic performance of EBOV IgG-indirect ELISAs based on antigens produced by classical virological and recombinant protein expression methods using human serum panels from EBOV non-infected and EBOV infected humans. abstract: Filovirus serological diagnosis and epidemiological investigations are hampered due to the unavailability of validated immunoassays. Diagnostic performance of three indirect enzyme-linked immunosorbent assays (I-ELISA) was evaluated for the detection of IgG antibody to Ebola virus (EBOV) in human sera. One I-ELISA was based on a whole EBOV antigen (WAg) and two utilized recombinant nucleocapsid (NP) and glycoproteins (GP), respectively. Validation data sets derived from individual sera collected in South Africa (SA), representing an EBOV non-endemic country, and from sera collected during an Ebola disease (EBOD) outbreak in Sierra Leone (SL), were categorized according to the compounded results of the three I-ELISAs and real time reverse-transcription polymerase chain reaction (RT-PCR). At the cut-off values selected at 95% accuracy level by the two-graph receiver operating characteristic analysis, specificity in the SA EBOV negative serum panel (n = 273) ranged from 98.17% (GP ELISA) to 99.27% (WAg ELISA). Diagnostic specificity in the SL EBOV negative panel (n = 676) was 100% by the three ELISAs. The diagnostic sensitivity in 423 RT-PCR confirmed EBOD patients was dependent on the time when the serum was collected after onset of disease. It significantly increased 2 weeks post-onset, reaching 100% sensitivity by WAg and NP and 98.1% by GP I-ELISA. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722596/ doi: 10.3390/v11080678 id: cord-317223-juw4xt8q author: Pedersen, Niels C. title: The causes of false-positives encountered during the screening of old-world primates for antibodies to human and simian retroviruses by ELISA date: 1986-11-30 words: 3923.0 sentences: 190.0 pages: flesch: 51.0 cache: ./cache/cord-317223-juw4xt8q.txt txt: ./txt/cord-317223-juw4xt8q.txt summary: False-positive ELISA antibody tests were particularly common among sera from mandrills, crab-eating macaques, lion-tailed macaques, African green monkeys, and DeBrazza''s and moustached guenons. False positive ELISA antibody tests, while sporadically encountered among most of the 50 species of oldworld primates, were especially prevalent in Mandrillus sphinx (mandrills), Macaca fasicularis (crab-eating macaques) , Macaca sifensus (lion-tailed macaques), Cercopithecus aethiops (African green monkeys), Cercopithecus neglectus (De-Brazza''s guenons), Cercopithecus cephus (moustached guenons) and Miopithecus talapoin (talapoins) . Specific antibodies to HTLV-III were found in 23 monkeys, 4/7 sooty mangabys, 11/15 talapoins, 2/11 False-positive ELISA antibody reactions to one or more viruses were found in a low proportion of individuals from many different species of old-world primates (Table 1) . Studies with African green monkey sera established the cause of false-positive ELISA antibody tests in this species; sera appeared to contain a specific antibody or antibodies that was directed against cell-associated protein(s) that were co-purified with the various virus preparations. abstract: Abstract Sera from 526 old-world primates representing 50 different species were screened by ELISA for antibodies to human T-lymphotropic viruses I and III, and simian retrovirus type 1 (SRV-1). About onefourth of the sera were positive by ELISA. There was a tendency, however, for the same sera to be positive for all three human and simian retroviruses. Only about one in five of the ELISA antibodypositive sera were confirmed to be positive by Western blotting. False-positive ELISA antibody tests were particularly common among sera from mandrills, crab-eating macaques, lion-tailed macaques, African green monkeys, and DeBrazza's and moustached guenons. Sera that were falsely positive in ELISA antibody tests to the three human and simian retroviruses were found to contain antibodies that reacted at comparable intensity with feline leukemia, infectious peritonitis and panleukopenia viruses. The false anti-viral activity of these sera was found to be due to antibodies that reacted with non-viral proteins that were copurified with all five virus preparations. These proteins were present in normal cat and human cells used to grow the various viruses and in gelatin. The implications of nonspecific cell-protein antibodies in primate sera were discussed in the light of this and previous seroepidemiologic studies of man and old-world monkeys. url: https://api.elsevier.com/content/article/pii/0166093486900236 doi: 10.1016/0166-0934(86)90023-6 id: cord-281760-34wuttqw author: Pereira, E.P.V. title: Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review date: 2019-05-22 words: 9686.0 sentences: 431.0 pages: flesch: 42.0 cache: ./cache/cord-281760-34wuttqw.txt txt: ./txt/cord-281760-34wuttqw.txt summary: Considering the fast development of IgY technology, this work aims to review its applications in human and animal health, in addition to increasing the potential use of these antibodies among researchers and consequently promoting the reduced use of non-invasive procedures on animals. extracted IgY from hens immunized with the recombinant protein FanC, from enterotoxigenic Escherichia coli (ETEC) and these antibodies bound specifically to FanC in ELISA, Western blot and Dot-blotting [59] , demanding, thus, more investigations to evaluate its viability as a potential immunotherapeutic compound. Anti-DENV2 IgY produced in goose was able to neutralize the virus in vitro and in vivo without binding to Fcγ receptors on myeloid cells and generating ADE (antibody dependent enhancement) in mice [57] . Hen egg yolk antibodies (IgY), production and use for passive immunization against bacterial enteric infections in chicken: a review Preventive effect of anti-VacA egg yolk immunoglobulin (IgY) on Helicobacter pylori-infected mice abstract: Egg yolk constitutes a relevant alternative source of antibodies. It presents some advantages over mammalian serum immunoglobulins regarding productivity, animal welfare and specificity. The main immunoglobulin present in avian blood (IgY) is transmitted to their offspring and accumulates in egg yolks, which enables the non-invasive harvesting of high amounts of antibodies. Moreover, due to structural differences and phylogenetic distance, IgY is more suitable for diagnostic purposes than mammalian antibodies, since it does not react with certain components of the human immune system and displays greater avidity for mammalian conserved proteins. IgY has been extensively used in health researches, as both therapeutic and diagnostic tool. This article aims to review its applications in both human and veterinary health. url: https://api.elsevier.com/content/article/pii/S1567576919302206 doi: 10.1016/j.intimp.2019.05.015 id: cord-290705-7xkt6u73 author: Petrini, Stefano title: Evaluation of Passive Immunity Induced by Immunisation Using Two Inactivated gE-deleted Marker Vaccines against Infectious Bovine Rhinotracheitis (IBR) in Calves date: 2020-01-04 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Different types of vaccines against Infectious Bovine Rhinotracheitis (IBR) are commercially available. Among these, inactivated glycoprotein E (gE)-deleted marker vaccines are commonly used, but their ability to induce passive immunity is poorly known. Here, we evaluated the passive immunity transferred from dams immunised with commercial inactivated gE-deleted marker vaccines to calves. We vaccinated 12 pregnant cattle devoid of neutralising antibodies against Bovine alphaherpesvirus 1 (BoHV-1) and divided them into two groups with 6 animals each. Both groups were injected with a different inactivated gE-deleted marker vaccine administrated via intranasal or intramuscular routes. An additional 6 pregnant cattle served as the unvaccinated control group. After calving, the number of animals in each group was increased by the newborn calves. In the dams, the humoral immune response was evaluated before calving and, subsequently, at different times until post-calving day 180 (PCD180). In addition, the antibodies in colostrum, milk, and in serum samples from newborn calves were evaluated at different times until PCD180. The results indicated that inactivated glycoprotein E (gE)-deleted marker vaccines are safe and produce a good humoral immune response in pregnant cattle until calving and PCD180. Moreover, results showed that, in calf serum, passive immunity persists until PCD180. url: https://doi.org/10.3390/vaccines8010014 doi: 10.3390/vaccines8010014 id: cord-348161-757c51xw author: Petrosova, A. title: Development of a highly sensitive, field operable biosensor for serological studies of Ebola virus in central Africa date: 2007-03-26 words: 5428.0 sentences: 264.0 pages: flesch: 48.0 cache: ./cache/cord-348161-757c51xw.txt txt: ./txt/cord-348161-757c51xw.txt summary: We employed a photo immobilization methodology based on a photoactivatable electrogenerated poly(pyrrole-benzophenone) film deposited upon an indium tin oxide (ITO) modified conductive surface fiber-optic. The photochemically modified optical fibers were tested as an immunosensor for detection of antibodies against Ebola virus, in animal and human sera, by use of a coupled chemiluminescent reaction. In this study we present a newly developed optical immunosensor for detection of antibodies to Ebola virus strains Zaire and Sudan, by using a photoimmobilization methodology based on a photoactivable electrogenerated polymer film. The optical fibers coated with poly(pyrrole-benzophenone) were soaked in diluted solution containing inactivated Ebola virus antigen (approximately 7.5 g/ml, the concentration was determined by Micro BCA Protein assay kit, PIERCE) and irradiated with UV light. The calibration curve obtained from the optical fiber immunosensor and ELISA for the detection of anti-Ebola subtype Zaire antibodies. abstract: We describe herein a newly developed optical immunosensor for detection of antibodies directed against antigens of the Ebola virus strains Zaire and Sudan. We employed a photo immobilization methodology based on a photoactivatable electrogenerated poly(pyrrole-benzophenone) film deposited upon an indium tin oxide (ITO) modified conductive surface fiber-optic. It was then linked to a biological receptor, Ebola virus antigen in this case, on the fiber tip through a light driven reaction. The photochemically modified optical fibers were tested as an immunosensor for detection of antibodies against Ebola virus, in animal and human sera, by use of a coupled chemiluminescent reaction. The immunosensor was tested for sensitivity, specificity, and compared to standard chemiluminescent ELISA under the same conditions. The analyte, anti-Ebola IgG, was detected at a low titer of 1:960,000 and 1:1,000,000 for subtypes Zaire and Sudan, respectively. While the same serum tested by ELISA was one order (24 times) less sensitive. url: https://doi.org/10.1016/j.snb.2006.07.005 doi: 10.1016/j.snb.2006.07.005 id: cord-317057-c2bwky6e author: Pickering, S. title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date: 2020-06-04 words: 5304.0 sentences: 269.0 pages: flesch: 53.0 cache: ./cache/cord-317057-c2bwky6e.txt txt: ./txt/cord-317057-c2bwky6e.txt summary: A highly specific in-house ELISA was developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs) on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. Accordingly, we developed a highly specific semi-quantitative 4 ELISA for the detection of anti-spike (S), -S receptor binding domain (RBD) and -N antibodies, and used this to cross-evaluate ten commercial antibody tests (seven lateral flow immunoassays (LFIAs), one chemiluminescent assay and two ELISAs) on a collection of 110 serum samples from confirmed RNA positive patients, and 50 pre-pandemic samples from March 2019. With no existing gold standard for the assessment of the serological response to SARS-CoV-2, we started by comparing commercial serological assays with the best performing configuration of the in-house ELISA (detection of anti-S IgM and IgG antibodies), which was also most likely to represent antibodies detected by the commercial tests. abstract: There is a clear requirement for an accurate SARS-CoV-2 antibody test, both as a complement to existing diagnostic capabilities and for determining community seroprevalence. We therefore evaluated the performance of a variety of antibody testing technologies and their potential as diagnostic tools. A highly specific in-house ELISA was developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays - a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs) - on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. There was a wide variation in the performance of the different platforms, with specificity ranging from 82% to 100%, and overall sensitivity from 60.9% to 87.3%. However, the head to head comparison of multiple serodiagnostic assays on identical sample sets revealed that performance is highly dependent on the time of sampling, with sensitivities of over 95% seen in several tests when assessing samples from more than 20 days post onset of symptoms. Furthermore, these analyses identified clear outlying samples that were negative in all tests, but were later shown to be from individuals with mildest disease presentation. Rigorous comparison of antibody testing platforms will inform the deployment of point of care technologies in healthcare settings and their use in the monitoring of SARS-CoV-2 infections. url: http://medrxiv.org/cgi/content/short/2020.06.02.20120345v1?rss=1 doi: 10.1101/2020.06.02.20120345 id: cord-351952-lhhjax3s author: Pickering, Suzanne title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings date: 2020-09-24 words: 5310.0 sentences: 247.0 pages: flesch: 48.0 cache: ./cache/cord-351952-lhhjax3s.txt txt: ./txt/cord-351952-lhhjax3s.txt summary: Highly specific in-house ELISAs were developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays—a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs)—on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. Accordingly, we developed a highly specific semi-quantitative ELISA for the detection of anti-spike (S), -S receptor binding domain (RBD) and -N antibodies, and used this to cross-evaluate ten commercial antibody tests (seven lateral flow immunoassays (LFIAs), one chemiluminescent assay and two ELISAs) on a collection of 110 serum samples from confirmed RNA positive patients, and 50 pre-pandemic samples from March 2019. With no existing standardised diagnostic test for the assessment of the serological response to SARS-CoV-2, we started by comparing commercial serological assays with the configuration of the in-house ELISA most likely to represent antibodies detected by the commercial tests (detection of anti-S IgM and IgG antibodies), and that had high specificity and sensitivity (S1 Table) . abstract: There is a clear requirement for an accurate SARS-CoV-2 antibody test, both as a complement to existing diagnostic capabilities and for determining community seroprevalence. We therefore evaluated the performance of a variety of antibody testing technologies and their potential use as diagnostic tools. Highly specific in-house ELISAs were developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assays—a chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs)—on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. There was a wide variation in the performance of the different platforms, with specificity ranging from 82% to 100%, and overall sensitivity from 60.9% to 87.3%. However, the head-to-head comparison of multiple sero-diagnostic assays on identical sample sets revealed that performance is highly dependent on the time of sampling, with sensitivities of over 95% seen in several tests when assessing samples from more than 20 days post onset of symptoms. Furthermore, these analyses identified clear outlying samples that were negative in all tests, but were later shown to be from individuals with mildest disease presentation. Rigorous comparison of antibody testing platforms will inform the deployment of point-of-care technologies in healthcare settings and their use in the monitoring of SARS-CoV-2 infections. url: https://doi.org/10.1371/journal.ppat.1008817 doi: 10.1371/journal.ppat.1008817 id: cord-257715-pbcr81qm author: Pignatelli, J. title: Molecular characterization of a new PToV strain. Evolutionary implications date: 2009-03-20 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Toroviruses are emergent viruses, belonging to the Nidovirales order, that remain mostly ignored, despite they are able to infect different species of domestic animals and humans, causing enteric diseases and diarrhea. Thus far, only five variants of porcine torovirus (PToV) have been identified. In this report we describe the identification and partial characterization of a new strain of porcine torovirus (PToV-BRES) that was detected by RT-PCR in a swine faecal specimen from a farm in Brescia (Italy). The complete genes coding for the nucleocapsid (N), hemagglutinin-esterase (HE) and membrane (M) proteins were amplified, and sequence analysis showed that PToV-BRES is a new PToV strain that, based on the HE gene sequence, is phylogenetically related to P4 strain, that was up to now the only member of a distinct PToV lineage. The nucleocapsid protein from PToV-BRES was expressed in insect cells as a his-tagged protein, purified by affinity chromatography and used to develop an ELISA method to detect antibodies against PToV. This assay was evaluated using a serum collection including 45 samples from three commercial farms from Spain. High antibody prevalence against PToV was observed in the three farms, both in adult animals and in piglets, which could suggest that PToV might be endemic in Spanish porcine population. The ELISA method developed in this work could be useful in future epidemiological surveys about toroviruses. url: https://doi.org/10.1016/j.virusres.2009.02.019 doi: 10.1016/j.virusres.2009.02.019 id: cord-334968-gonx5taq author: Pignatelli, Jaime title: Lineage specific antigenic differences in porcine torovirus hemagglutinin-esterase (PToV-HE) protein date: 2013-12-23 words: 6772.0 sentences: 291.0 pages: flesch: 50.0 cache: ./cache/cord-334968-gonx5taq.txt txt: ./txt/cord-334968-gonx5taq.txt summary: In the first case, the two PToV-HE lineages were detected even within the same animal at two sequential sampling time points, indicating that both PToV strains carrying different HE proteins coexisted on the same farm infecting the same piglets, and suggesting that the immune response generated against one PToV strain did not protect the animals against the infection by the other strain. Hence, in order to examine the potential existence of antigenic differences between the two lineages of PToV-HE proteins, piglet serum samples were analyzed with an HI assay to detect the presence of specific antibodies against the receptor binding domain that prevents the RBC hemagglutination. In addition, to study the antibody response against the whole protein by a different approach, soluble c-myc tagged HE52.7 and HE52.11 proteins were generated by rVV methodology to obtain highly purified coating antigens that were used in ELISA to test the same field serum samples. abstract: Hemagglutinin-esterases (HE) are viral envelope proteins present in some members from the toro-, corona- and orthomyxovirus families, all related with enteric and/or respiratory tract infections. HE proteins mediate reversible binding to sialic acid receptor determinants, very abundant glycan residues in the enteric and respiratory tracts. The role of the HE protein during the torovirus infection cycle remains unknown, although it is believed to be important in the natural infection process. The phylogenetic analysis of HE coding sequences from porcine torovirus (PToV) field strains revealed the existence of two distinct HE lineages. In a previous study, PToV virus strains with HE proteins from the two lineages were found coexisting in a pig herd, and they were even obtained from the same animal at two consecutive sampling time points. In this work, we report antigenic differences between the two HE lineages, and discuss the possible implications that the coexistence of viruses belonging to both lineages might have on the spread and sustainment of PToV infection in the farms. url: https://www.ncbi.nlm.nih.gov/pubmed/24364900/ doi: 10.1186/1297-9716-44-126 id: cord-306390-pzzev8hd author: Reisinger, Emil C. title: Mütter-Screening in einem COVID-19-Niedrig-Pandemiegebiet: Bestimmung SARS-CoV-2-spezifischer Antikörper bei 401 Rostocker Müttern mittels ELISA und Immunfluoreszenz-Bestätigungstest date: 2020-06-22 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Background In children, the infection with SARS-CoV-2, the cause of COVID-19, tends to be clinically inapparent more often or less severe than in adults. The spread of this infection from children poses a danger to vulnerable adults. Therefore, child care institutions and schools currently are widely closed. Methods Since the status of infection tends to be congruent in mothers and their children, we tested 401 mothers of children between 1 and 10 years in the city of Rostock (State of Mecklenburg-Westpomerania, northeast of Germany), for the presence of RNA of SARS-CoV-2 in throat swabs, and of antibodies against SARS-CoV-2 in serum. Results In none of the mothers tested, RNA of this virus was detected in the throat swab. In the ELISA test, IgG antibodies were positive in one serum sample, IgA antibodies were positive in 11, and borderline in 3 serum samples. All 401 sera were negative in the indirect immunofluorescence test (IIFT) with FITC-labeled IgG, IgA, und IgM antibodies. Conclusion At the time of this study, neither SARS-CoV-2 RNA, nor specific antibodies against SARS-CoV-2 were detectable in the mothers tested in Rostock. url: https://www.ncbi.nlm.nih.gov/pubmed/32572869/ doi: 10.1055/a-1197-4293 id: cord-287590-jjft3den author: Rodák, L. title: Use of Monoclonal Antibodies in Blocking ELISA Detection of Transmissible Gastroenteritis Virus in Faeces of Piglets date: 2005-05-05 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Monoclonal antibodies (mAb) to the transmissible gastroenteritis virus (TGEV) nucleoprotein (N) and membrane protein (M) were prepared and used for the comparative assessment of three blocking ELISA variants to detect TGEV. The competitive blocking ELISA format showed the highest sensitivity, allowing detection of 10(3) TCID(50) TGEV/ml in culture medium. Ninety‐nine porcine field faecal samples obtained from 37 herds affected with diarrhoea were examined, and various TGEV levels were found in nine samples from six herds. However, only in three samples were significant TGEV concentrations demonstrated. The relationship between incidence of TGEV gastroenteritis and the spread of porcine respiratory coronavirus infection in pig farms is discussed. url: https://www.ncbi.nlm.nih.gov/pubmed/15876221/ doi: 10.1111/j.1439-0450.2005.00829.x id: cord-322529-3xn5v54s author: Rodák, L. title: Verification of Sensitivity and Specificity of Group A Rotavirus Detection in Piglets Faeces with Monoclonal Blocking ELISA Methods date: 2004-06-30 words: 3215.0 sentences: 196.0 pages: flesch: 48.0 cache: ./cache/cord-322529-3xn5v54s.txt txt: ./txt/cord-322529-3xn5v54s.txt summary: title: Verification of Sensitivity and Specificity of Group A Rotavirus Detection in Piglets Faeces with Monoclonal Blocking ELISA Methods Selected competitive blocking ELISA (CB‐ELISA) and electron microscopy (EM) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. The sensitivity and specificity of the CB‐ELISA was verified both by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhoea virus (PEDV) in each analysis and by comparative examination of samples with the commercial ELISA kit. Sensitivity comparison of three variants of the blocking ELISA method of rotavirus A detection were performed by box titrations in microtitre plate wells pre-coated with binding antibodies. Sensitivity of the CB-ELISA method and DAS-ELISA kit was compared by examination of faecal sample of experimentally infected piglet twofold diluted 2· to 1024·. By examination of positive faecal sample twofold diluted 2· to 1024·, at least 10 times higher sensitivity of CB-ELISA method was demonstrated (Table 2) . abstract: Monoclonal antibodies to group A rotavirus Vp6 protein were prepared and used for verification of three blocking enzyme‐linked immunosorbent assay (ELISA) modifications to detect rotavirus A. Selected competitive blocking ELISA (CB‐ELISA) and electron microscopy (EM) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. Rotavirus was detected in 43 samples (22.2%) by CB‐ELISA method, whereas in 26 (13.4%) samples by EM examination. However, of 26 samples positive by EM, rotavirus A was detected by CB‐ELISA in 19 (73.1%) samples; indicating the share of group A rotavirus in all cases of gastroenteritis caused by rotavirus. The sensitivity and specificity of the CB‐ELISA was verified both by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhoea virus (PEDV) in each analysis and by comparative examination of samples with the commercial ELISA kit. The CB‐ELISA sensitivity was positively affected by examination of samples in the presence of chelating agent. url: https://www.ncbi.nlm.nih.gov/pubmed/15228549/ doi: 10.1111/j.1439-0450.2004.00746.x id: cord-031190-4qpnlgb5 author: Sahu, Kamal K title: Current Perspectives on Diagnostic Assays and Anti-PF4 Antibodies for the Diagnosis of Heparin-Induced Thrombocytopenia date: 2020-08-17 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Heparin-induced thrombocytopenia (HIT) is a recognized clinical entity in patients receiving unfractionated heparin and low–molecular weight heparin. Currently, diagnosing HIT includes the combination of a physician’s clinical suspicion based on a clinical scoring system and a series of laboratory tests. In the present article, we discuss challenges in suspecting and diagnosing HIT in consideration of the turnaround time of available tests and recent advances in techniques and methodologies of newer immunoassays and functional assays. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7443028/ doi: 10.2147/jbm.s232648 id: cord-003623-n01rgqyv author: Schuh, Amy J. title: Comparative analysis of serologic cross-reactivity using convalescent sera from filovirus-experimentally infected fruit bats date: 2019-04-30 words: 6363.0 sentences: 277.0 pages: flesch: 37.0 cache: ./cache/cord-003623-n01rgqyv.txt txt: ./txt/cord-003623-n01rgqyv.txt summary: To evaluate the ability of our system comprising seven filovirus-specific indirect ELISAs to predict the filovirus species most antigenically similar to the species responsible for past infection, we tested seven Marburg virus convalescent serum 35 or whole blood 36 samples collected from experimentally inoculated ERBs. Five of these samples www.nature.com/scientificreports www.nature.com/scientificreports/ were collected four weeks post primary Marburg virus inoculation 35, 36 , while two of the samples were collected at 23 and 27 weeks post primary inoculation following a "natural" boost (i.e., Marburg virus-specific antibody levels waned in these bats and then increased following contact with infectious cagemates) 36 . Although significant levels of serological IgG cross-reactivity were observed between the prime-boost filovirus-specific antisera and some of the filovirus antigens, when the overall covariance of the seven-individual indirect ELISAs in the system were considered, we were able to predict the filovirus species responsible for past infection 100% of the time using as little as 25 μL of sera (each serum was tested against each antigen in duplicate). abstract: With the exception of Reston and Bombali viruses, the marburgviruses and ebolaviruses (family Filoviridae) cause outbreaks of viral hemorrhagic fever in sub-Saharan Africa. The Egyptian rousette bat (ERB) is a natural reservoir host for the marburgviruses and evidence suggests that bats are also natural reservoirs for the ebolaviruses. Although the search for the natural reservoirs of the ebolaviruses has largely involved serosurveillance of the bat population, there are no validated serological assays to screen bat sera for ebolavirus-specific IgG antibodies. Here, we generate filovirus-specific antisera by prime-boost immunization of groups of captive ERBs with all seven known culturable filoviruses. After validating a system of filovirus-specific indirect ELISAs utilizing infectious-based virus antigens for detection of virus-specific IgG antibodies from bat sera, we assess the level of serological cross-reactivity between the virus-specific antisera and heterologous filovirus antigens. This data is then used to generate a filovirus antibody fingerprint that can predict which of the filovirus species in the system is most antigenically similar to the species responsible for past infection. Our filovirus IgG indirect ELISA system will be a critical tool for identifying bat species with high ebolavirus seroprevalence rates to target for longitudinal studies aimed at establishing natural reservoir host-ebolavirus relationships. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6491471/ doi: 10.1038/s41598-019-43156-z id: cord-311811-nrodyagi author: Schutzer, Steven E. title: The use of host factors in microbial forensics date: 2019-12-06 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Advances have been made in the forensic analysis of microbes and toxins. An underdeveloped and underutilized area in microbial forensics is how the host interacts with microorganisms in a way that provides unique signatures for forensic use. For forensic purposes, an immediate goal is to distinguish a potential victim and innocent person from a perpetrator, and to distinguish between a naturally acquired or intentional infection. Principal methods that are sufficiently developed are characterization of the humoral immune response to microbial antigens including vaccine-induced immunity and detection of antibiotics that may be present in a possible perpetrator. This chapter presents central elements of the host response in a simplified fashion and describes a representative example, which, in the appropriate context, has a high potential of providing evidence that may aid an investigation to distinguish a perpetrator from a victim. This chapter also presents information about the immune system so that the interested reader can have a fuller understanding of the immune response in general. url: https://www.sciencedirect.com/science/article/pii/B9780128153796000143 doi: 10.1016/b978-0-12-815379-6.00014-3 id: cord-281101-gv1sgbk1 author: Shin, Gu-Choul title: Preparation and characterization of a novel monoclonal antibody specific to severe acute respiratory syndrome-coronavirus nucleocapsid protein date: 2006-08-30 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Severe acute respiratory syndrome-coronavirus nucleocapsid (SARS-CoV N) protein has been found to be important to the processes related to viral pathogenesis, such as virus replication, interference of the cell process and modulation of host immune response; detection of the antigen has been used for the early diagnosis of infection. We have used recombinant N protein expressed in insect cells to generate 17 mAbs directed against this protein. We selected five mAbs that could be used in various diagnostic assays, and all of these mAbs recognized linear epitopes. Three IgG(2b) mAbs were recognized within the N-terminus of N protein, whereas the epitope of two IgG(1) mAbs localized within the C-terminus. These mAbs were found to have significant reactivity with both non-phosphorylated and phosphorylated N proteins, which resulted in high reactivity with native N protein in virus-infected cells; however, they did not show cross-reactivity with human coronavirus. Therefore, these results suggested that these mAbs would be useful in the development of various diagnostic kits and in future studies of SARS-CoV pathology. url: https://www.ncbi.nlm.nih.gov/pubmed/16942813/ doi: 10.1016/j.virusres.2006.07.004 id: cord-312456-6lxc2rj2 author: Soltan, Mohamed A. title: Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species date: 2016-05-11 words: 4209.0 sentences: 218.0 pages: flesch: 53.0 cache: ./cache/cord-312456-6lxc2rj2.txt txt: ./txt/cord-312456-6lxc2rj2.txt summary: title: Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species The aim of this investigation was to evaluate a commercially available RT-iiPCR assay for RVA detection in feces from different animal species. Therefore, the aim of our investigation was to evaluate a recently available insulated isothermal RT-PCR (RT-iiPCR) reagent set (POCKIT TM Rotavirus A Reagent Set, GeneReach USA, Lexington, MA, USA) with use of a portable PCR machine, which could potentially be used for point-of-need detection for RVA in the feces of different animal species. Additionally, the sensitivity of the rotavirus RT-iiPCR reagent set was evaluated by comparison with the commercially available rtRT-PCR assay using 10-fold serial dilutions of nucleic acid extracted from a positive bovine clinical sample. There was a significant difference in the number of positive samples detected with the in-house rtRT-PCR assay versus the other two molecular tests. abstract: There is no gold standard for detection of Rotavirus Group A (RVA), one of the main causes of diarrhea in neonatal animals. Sensitive and specific real-time RT-PCR (rtRT-PCR) assays are available for RVA but require submission of the clinical samples to diagnostic laboratories. Patient-side immunoassays for RVA protein detection have shown variable results, particularly with samples from unintended species. A sensitive and specific test for detection of RVA on the farm would facilitate rapid management decisions. The insulated isothermal RT-PCR (RT-iiPCR) assay works in a portable machine to allow sensitive and specific on-site testing. The aim of this investigation was to evaluate a commercially available RT-iiPCR assay for RVA detection in feces from different animal species. This assay was compared to an in-house rtRT-PCR assay and a commercially available rtRT-PCR kit, as well as an ELISA and EM for RVA detection. All three PCR assays targeted the well-conserved NSP5 gene. Clinical fecal samples from 108 diarrheic animals (mainly cattle and horses) were tested. The percentage of positive samples by ELISA, EM, in-house rtRT-PCR, commercial rtRT-PCR, and RT-iiPCR was 29.4%, 31%, 36.7%, 51.4%, 56.9%, respectively. The agreement between different assays was high (81.3–100%) in samples containing high viral loads. The sensitivity of the RT-iiPCR assay appeared to be higher than the commercially available rtRT-PCR assay, with a limit of detection (95% confidence index) of 3–4 copies of in vitro transcribed dsRNA. In conclusion, the user-friendly, field-deployable RT-iiPCR system holds substantial promise for on-site detection of RVA. url: https://www.ncbi.nlm.nih.gov/pubmed/27180038/ doi: 10.1016/j.jviromet.2016.05.006 id: cord-293770-n4ooziv4 author: Soykut, Esra Acar title: Selection of staphylococcal enterotoxin B (SEB)-binding peptide using phage display technology date: 2008-05-23 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: In this study, peptides were selected to recognize staphylococcal enterotoxin B (SEB) which cause food intoxication and can be used as a biological war agent. By using commercial M13 phage library, single plaque isolation of 38 phages was done and binding affinities were investigated with phage-ELISA. The specificities of the selected phage clones showing high affinity to SEB were checked by using different protein molecules which can be found in food samples. Furthermore, the affinities of three selected phage clones were determined by using surface plasmon resonance (SPR) sensors. Sequence analysis was realized for three peptides showing high binding affinity to SEB and WWRPLTPESPPA, MNLHDYHRLFWY, and QHPQINQTLYRM amino acid sequences were obtained. The peptide sequence with highest affinity to SEB was synthesized with solid phase peptide synthesis technique and thermodynamic constants of the peptide–SEB interaction were determined by using isothermal titration calorimetry (ITC) and compared with those of antibody–SEB interaction. The binding constant of the peptide was determined as 4.2 ± 0.7 × 10(5) M(−1) which indicates a strong binding close to that of antibody. url: https://api.elsevier.com/content/article/pii/S0006291X08004993 doi: 10.1016/j.bbrc.2008.03.065 id: cord-261372-xjbs09gi author: Sozzi, Enrica title: Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus date: 2010-02-28 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Abstract Porcine epidemic diarrhoea (PED) is a contagious enteric disease of pigs caused by a coronavirus. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) based on the use of monoclonal antibodies was developed for the detection of porcine epidemic diarrhoea virus (PEDV). The DAS-ELISA was compared with RT-PCR in the examination of 506 specimens collected during 2006–2007 from pigs originating from different farms located in the Po valley. Both faecal samples obtained directly from the rectum of live animals showing clinical signs and intestinal samples collected from the caecum of deceased pigs were included in the study. The correlation between the two methods was higher when testing faecal samples (K =0.97, 95% CI: 0.94–1.00) than testing intestinal samples (K =0.62, 95% CI: 0.35–0.89). The use of ELISA technology provided an efficient and effective mean of evaluating the presence of coronavirus PED antigen in field samples and indicates that this procedure is a very useful tool in epidemiological studies. url: https://api.elsevier.com/content/article/pii/S0034528809001362 doi: 10.1016/j.rvsc.2009.05.009 id: cord-334165-7gfk554m author: Stadlbauer, Daniel title: SARS‐CoV‐2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup date: 2020-04-17 words: 5286.0 sentences: 414.0 pages: flesch: 69.0 cache: ./cache/cord-334165-7gfk554m.txt txt: ./txt/cord-334165-7gfk554m.txt summary: Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two‐stage ELISA for high‐throughput screening of human serum samples for antibodies binding to the spike protein of SARS‐CoV‐2 We reported in our earlier work that individuals not exposed to SARS-CoV-2 are completely naïve to the spike protein, and their serum samples show little or no reactivity in an ELISA (Amanat et al., 2020) . We developed this as a two-stage ELISA in which the first stage (''a'' steps below) includes relatively high-throughput screening of samples in a single serum dilution against the RBD (which expresses very well and therefore can be produced in greater quantities). This is followed by a second stage (''b'' steps below) in which positive samples from the first stage undergo a confirmatory ELISA against the full-length spike protein (which is harder to express; therefore there is usually less available). i. Thaw the required number of vials of antigen (SARS-CoV-2 RBD protein) to coat 96-well microtiter ELISA plates at a concentration of 2 μg/ml. abstract: In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS‐CoV‐2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re‐infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS‐CoV‐2 that can serve as a substrate for immunological assays, as well as a two‐stage serological enzyme‐linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories. © 2020 The Authors. Current Protocols in Microbiology published by Wiley Periodicals LLC. Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two‐stage ELISA for high‐throughput screening of human serum samples for antibodies binding to the spike protein of SARS‐CoV‐2 url: https://doi.org/10.1002/cpmc.100 doi: 10.1002/cpmc.100 id: cord-317123-0tdfvlqd author: Tan, Xiaotian title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples date: 2020-04-21 words: 4154.0 sentences: 220.0 pages: flesch: 52.0 cache: ./cache/cord-317123-0tdfvlqd.txt txt: ./txt/cord-317123-0tdfvlqd.txt summary: Here, we present a microfluidic ELISA technology for rapid (15-20 minutes), quantitative, sensitive detection of SARS-CoV-2 biomarkers using SARS-CoV-2 specific IgG and viral antigen – S protein in serum. We also characterized various humanized monoclonal IgG, and identified a candidate with a high binding affinity towards SARS-CoV-2 S1 protein that can serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. In this work, we present a microfluidic ELISA technology for rapid (15-20 minutes), quantitative, and sensitive detection of SARS-CoV-2 biomarkers using SARS-CoV-2 specific IgG and viral antigen -S protein, both of which are spiked in serum, as a model system. For the anti-S1 IgG detection experiments (see Figure S2 (B) for the detailed protocol), various concentrations of monoclonal antibodies were prepared by diluting the stock solutions with 50 times diluted human serum (the serum was diluted with 1× reagent diluent, which correlates to 1% BSA). abstract: COVID-19 pandemic has caused tens of thousands of deaths and is now a severe threat to global health. Clinical practice has demonstrated that the SARS-CoV-2 S1 specific antibodies and viral antigens can be used as diagnostic and prognostic markers of COVID-19. However, the popular point-of-care biomarker detection technologies, such as the lateral-flow test strips, provide only yes/no information and have very limited sensitivities. Thus, it has a high false negative rate and cannot be used for the quantitative evaluation of patient’s immune response. Conventional ELISA (enzyme-linked immunosorbent assay), on the other hand, can provide quantitative, accurate, and sensitive results, but it involves complicated and expensive instruments and long assay time. In addition, samples need to be sent to centralized labs, which significantly increases the turn-around time. Here, we present a microfluidic ELISA technology for rapid (15-20 minutes), quantitative, sensitive detection of SARS-CoV-2 biomarkers using SARS-CoV-2 specific IgG and viral antigen – S protein in serum. We also characterized various humanized monoclonal IgG, and identified a candidate with a high binding affinity towards SARS-CoV-2 S1 protein that can serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. Furthermore, we demonstrated that our microfluidic ELISA platform can be used for rapid affinity evaluation of monoclonal anti-S1 antibodies. The microfluidic ELISA device is highly portable and requires less than 10 μL of samples for each channel. Therefore, our technology will greatly facilitate rapid and quantitative analysis of COVID-19 patients and vaccine recipients at point-of-care. url: https://doi.org/10.1101/2020.04.20.052233 doi: 10.1101/2020.04.20.052233 id: cord-326232-668f53qc author: Toftaker, Ingrid title: Evaluation of a multiplex immunoassay for bovine respiratory syncytial virus and bovine coronavirus antibodies in bulk tank milk against two indirect ELISAs using latent class analysis date: 2018-06-01 words: 5627.0 sentences: 307.0 pages: flesch: 56.0 cache: ./cache/cord-326232-668f53qc.txt txt: ./txt/cord-326232-668f53qc.txt summary: The objective of this study was to estimate sensitivity and specificity across different cut-off values for the MVD-Enferplex BCV/BRSV multiplex, by comparing them to a commercially available ELISA, the SVANOVIR(®) BCV-Ab and SVANOVIR(®) BRSV-Ab, respectively. The aim of this study was to estimate the test sensitivity and specificity of the newly developed MVD-Enferplex BCV/BRSV multiplex across different cut-off values, for detection of antibodies in BTM. Estimates of test parameters and true prevalence in the two subpopulations across different cut-off values for the BCV multiplex and the BCV ELISA are presented in Table 5 . The chosen cut-off will Table 4 Results from the sensitivity analysis (BRSV): Median estimates and 95% posterior credibility intervals (PCI) of the sensitivity (Se) and specificity (Sp) of bulk tank milk BRSV multiplex and BRSV ELISA at the manufacturers'' recommended cut-off (alternative 2, Fig. 1 ), for the conditionally independent (CID) model and conditionally dependent (COC) models where the covariance is expressed as proportions of maximum possible value. abstract: Bovine respiratory syncytial virus (BRSV) and bovine coronavirus (BCV) are responsible for respiratory disease and diarrhea in cattle worldwide. The Norwegian control program against these infections is based on herd-level diagnosis using a new multiplex immunoassay. The objective of this study was to estimate sensitivity and specificity across different cut-off values for the MVD-Enferplex BCV/BRSV multiplex, by comparing them to a commercially available ELISA, the SVANOVIR(®) BCV-Ab and SVANOVIR(®) BRSV-Ab, respectively. We analyzed bulk tank milk samples from 360 herds in a low- and 360 herds in a high-prevalence area. As none of the tests were considered perfect, estimation of test characteristics was performed using Bayesian latent class models. At the manufacturers’ recommended cut-off values, the median sensitivity for the BRSV multiplex and the BRSV ELISA was 94.4 [89.8–98.7 95% Posterior Credibility Interval (PCI)] and 99.8 [98.7–100 95% PCI], respectively. The median specificity for the BRSV multiplex was 90.6 [85.5–94.4 95% PCI], but only 57.4 [50.5–64.4 95% PCI] for the BRSV ELISA. However, increasing the cut-off of the BRSV ELISA increased specificity without compromising sensitivity. For the BCV multiplex we found that by using only one of the three antigens included in the test, the specificity increased, without concurrent loss in sensitivity. At the recommended cut-off this resulted in a sensitivity of 99.9 [99.3–100 95% PCI] and specificity of 93.7 [88.8–97.8 95% PCI] for the multiplex and a sensitivity of 99.5 [98.1–100 95% PCI] and a specificity of 99.6 [97.6–100 95% PCI] for the BCV ELISA. url: https://api.elsevier.com/content/article/pii/S0167587717307961 doi: 10.1016/j.prevetmed.2018.03.008 id: cord-026559-xx52u01h author: Tripathi, Siddhartha title: Blood Plasma Microfluidic Device: Aiming for the Detection of COVID-19 Antibodies Using an On-Chip ELISA Platform date: 2020-06-10 words: 2010.0 sentences: 125.0 pages: flesch: 52.0 cache: ./cache/cord-026559-xx52u01h.txt txt: ./txt/cord-026559-xx52u01h.txt summary: title: Blood Plasma Microfluidic Device: Aiming for the Detection of COVID-19 Antibodies Using an On-Chip ELISA Platform We propose to first separate plasma from whole human blood using a microfluidic device and subsequently perform the detection of antibodies in the separated plasma using a semi-automated on-chip ELISA. The reported plasma separation microdevice is not only an alternate to the centrifuge, but it can also be easily integrated with a biosensing platform/detection technology (for example, ELISA) and result in a point-of-care device. (2020) have reported successful detection of SARS-CoV-2 virus with high sensitivity in swab specimens Fig. 1 a Blood plasma microdevice design and zoomed view at the junction. Herein, we propose the integration of sandwich ELISA (enzyme-linked immunoabsorbent assay) with the blood plasma separation microdevice to detect COVID-19 antibodies after minor modifications in the design. b Experi-mental sandwich ELISA: showing steps to identify the presence of SARS-CoV-2 (COVID-19) antibodies present in blood plasma separated and flows towards the plasma outlet reservoir. abstract: COVID-19 is a public health emergency of international concern. Detection of SARS-CoV-2 virus is an important step towards containing the virus spread. Although viral detection using molecular diagnostic methods is quite common and efficient, these methods are prone to errors, laborious and time consuming. There is an urgent need for blood-based tests which are simple to use, accurate, less time consuming, portable and cost-effective. Human blood plasma contains water, proteins, organic and in-organic substances including bacteria and viruses. Blood plasma can be effectively used to detect COVID-19 antibodies. The immune system generates antibodies (IgM/IgG proteins) in response to the virus and identification of these antibodies is related to the presence of the infection in the patient in the past. Therefore, detecting and testing the presence of these antibodies will be extremely useful for monitoring and surveillance of the population (Petherick, Lancet 395:1101–1102, 2020). Herein, we describe and propose a microfluidic ELISA (enzyme-linked immunosorbent assay) system to detect COVID-19 antibodies on a lab-on-chip platform. We propose to first separate plasma from whole human blood using a microfluidic device and subsequently perform the detection of antibodies in the separated plasma using a semi-automated on-chip ELISA. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7283038/ doi: 10.1007/s41403-020-00123-9 id: cord-022353-q2k2krnm author: W. Quimby, Fred title: Clinical Chemistry of the Laboratory Mouse date: 2007-09-02 words: 30195.0 sentences: 1702.0 pages: flesch: 48.0 cache: ./cache/cord-022353-q2k2krnm.txt txt: ./txt/cord-022353-q2k2krnm.txt summary: Assessment of long-term average blood glucose levels in mice is also available by RIAs measuring glycosylated hemoglobin and glycosylated serum proteins (collectively known as fructosamines) (Gould et al. Leptin resistance, a common feature of obesity in mice and humans, has also been shown to result, in part, from the shedding of membrane-bound hepatic leptin receptors into the plasma, where soluble receptors modulate circulating leptin levels and possibly its biologic activity (Cohen et al. d. OTHER ANALYTES ASSOCIATED WITH LIPID METABOLISM AND ATHEROSCLEROSIS IN MICE ELISA kits are commercially available for the quantitation of many mouse coagulation proteins including: fibrinogen, factor VII, d-dimer, tissue factor, and von Willebrand''s factor antigen. The ability of the first component of complement, C1, to bind specific sites on the heavy chain of mouse IgG2b and activate a sequence of reactions leading to production of a molecular unit capable of lysing a target cell membrane has established the complement system as the primary mediator of antibody-antigen reactions. abstract: The frontier of clinical chemistry in the mouse has advanced and expanded because of two major events such as, the increasing reliance on mice in biomedical research, and increasing availability of practical yet sophisticated techniques and instrumentations that have allowed for the detection of a wider variety of biomarkers of disease. The progression of these two events is partially driven by the increasing regulatory demands related to safety/toxicity assessment of novel drug development. The availability of inbred strains has led to major breakthroughs in cancer, biology, and immunology. In addition, outbred stocks continue to be utilized in a wide variety of studies but particularly in the fields of toxicology and pharmacology. The power of these models to elucidate the genetic basis of disease cannot be overemphasized. This provided complete nucleotide sequences for each genome allowing investigators to quickly develop the equivalent murine model for many of the inherited human diseases. Transgenic and knockout mice have helped clarify disease pathogenesis in virtually every area of medicine and often elucidated biochemical pathways, previously unknown, which are now subject to testing and quantification. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7155603/ doi: 10.1016/b978-012369454-6/50060-1 id: cord-318444-sgm24q1i author: Walter, Justin D. title: Sybodies targeting the SARS-CoV-2 receptor-binding domain date: 2020-05-16 words: 5902.0 sentences: 416.0 pages: flesch: 49.0 cache: ./cache/cord-318444-sgm24q1i.txt txt: ./txt/cord-318444-sgm24q1i.txt summary: Two independently prepared RBD constructs were used for in vitro sybody selections, and resulting single clones that could bind the full spike ectodomain were sequenced, expressed, and purified. Six unique sybodies show favorable binding affinity to the SARS-CoV-2 spike, and five of these were also found to substantially attenuate the interaction between the viral RBD and human ACE2. While this purified pre-fusion spike (PFS) had not yet been available for binder selections and characterization by grating-coupled interferometry, it was used to conduct ELISAs in order to identify selected sybodies which recognize the RBD in the pre-fusion context (see below). Since virulence of SARS-CoV-2 is dependent on the ability of the viral RBD to bind to human ACE2 (hACE2), we sought to determine which of the 57 selected sybodies that were well-behaved upon purification could inhibit interaction between the isolated RBD and purified hACE2. abstract: The COVID-19 pandemic, caused by the novel coronavirus SARS-CoV-2, has resulted in a global health and economic crisis of unprecedented scale. The high transmissibility of SARS-CoV-2, combined with a lack of population immunity and prevalence of severe clinical outcomes, urges the rapid development of effective therapeutic countermeasures. Here, we report the generation of synthetic nanobodies, known as sybodies, against the receptor-binding domain (RBD) of SARS-CoV-2. In an expeditious process taking only twelve working days, sybodies were selected entirely in vitro from three large combinatorial libraries, using ribosome and phage display. We obtained six strongly enriched sybody pools against the isolated RBD and identified 63 unique anti-RBD sybodies which also interact in the context of the full-length SARS-CoV-2 spike ectodomain. Among the selected sybodies, six were found to bind to the viral spike with double-digit nanomolar affinity, and five of these also showed substantial inhibition of RBD interaction with human angiotensin-converting enzyme 2 (ACE2). Additionally, we identified a pair of anti-RBD sybodies that can simultaneously bind to the RBD. It is anticipated that compact binders such as these sybodies could feasibly be developed into an inhalable drug that can be used as a convenient prophylaxis against COVID-19. Moreover, generation of polyvalent antivirals, via fusion of anti-RBD sybodies to additional small binders recognizing secondary epitopes, could enhance the therapeutic potential and guard against escape mutants. We present full sequence information and detailed protocols for the identified sybodies, as a freely accessible resource. url: https://doi.org/10.1101/2020.04.16.045419 doi: 10.1101/2020.04.16.045419 id: cord-278324-eqqvwwh6 author: Wang, Huanan title: Development of a sensitive and specific xMAP assay for detection of antibodies against infectious laryngotracheitis and bronchitis viruses date: 2018-09-21 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination. METHOD: The microspheres coated with purified recombinant glycoprotein D (gD) of ILTV or nucleocapsid (N) protein of IBV were incubated with serum samples. The simultaneous quantification of ILTV and IBV antibodies were achieved through the interrogation of microspheres by Luminex 200 detection system. . RESULTS: This xMAP detection demonstrated no nonspecific reactions with avian influenza virus (AIV), avian leukosis virus (ALV), newcastle disease virus (NDV), and Marek’s disease virus (MDV). The results also demonstrated that the xMAP assay was four times more sensitive than the enzyme-linked immunosorbent assay (ELISA) for ILTV detection and two times more sensitive for IBV detection. A total of 90 chicken serum samples from a chicken farm were tested by xMAP and ELISA assays. The results showed that the coincidence rates were 84.44 and 100% for ILTV and IBV detection, respectively. CONCLUSION: This study exhibited an opportunity for the differential diagnosis through simultaneous detection of multiplex antibodies in serum and can be used for the multiplex antibodies evaluation after vaccination. url: https://doi.org/10.1186/s12985-018-1048-x doi: 10.1186/s12985-018-1048-x id: cord-262268-gm99cadh author: Wang, Jingqiang title: Assessment of Immunoreactive Synthetic Peptides from the Structural Proteins of Severe Acute Respiratory Syndrome Coronavirus date: 2003-12-01 words: 4027.0 sentences: 189.0 pages: flesch: 49.0 cache: ./cache/cord-262268-gm99cadh.txt txt: ./txt/cord-262268-gm99cadh.txt summary: Consequently, we thoroughly investigated the immunoreactivities with patient sera of a series of synthesized peptides from SARS-coronavirus structural proteins. Results: Four epitopic sites, S599, M137, N66, and N371-404, located in the SARS-coronavirus S, M, and N proteins, respectively, were detected by screening synthesized peptides. The peptides representing the COOH terminus of the N protein, in particular N371 and N385, had high absorbance/cutoff value ratios with the highest positive detection rate and the lowest hydrophobicity score among all of the synthesized peptides (Fig. 1C, and Fig. 3 in the online Data Supplement). The other 17 peptides reacted only slightly with the sera from SARS patients and gave low detection rates, suggesting that the regions of the S protein covered by these peptides have no epitopic site. The patient sera preincubated with 4 mg/L S599 or N385 gave a 25-30% lower response in the ELISA (data not shown), suggesting that the two peptides could compete with SARS coronavirus for binding to the antibodies in SARS serum. abstract: Background: The widespread threat of severe acute respiratory syndrome (SARS) to human life has spawned challenges to develop fast and accurate analytical methods for its early diagnosis and to create a safe antiviral vaccine for preventive use. Consequently, we thoroughly investigated the immunoreactivities with patient sera of a series of synthesized peptides from SARS-coronavirus structural proteins. Methods: We synthesized 41 peptides ranging in size from 16 to 25 amino acid residues of relatively high hydrophilicity. The immunoreactivities of the peptides with SARS patient sera were determined by ELISA. Results: Four epitopic sites, S599, M137, N66, and N371-404, located in the SARS-coronavirus S, M, and N proteins, respectively, were detected by screening synthesized peptides. Notably, N371 and N385, located at the COOH terminus of the N protein, inhibited binding of antibodies to SARS-coronavirus lysate and bound to antibodies in >94% of samples from SARS study patients. N385 had the highest affinity for forming peptide-antibody complexes with SARS serum. Conclusions: Five peptides from SARS structural proteins, especially two from the COOH terminus of the N protein, appear to be highly immunogenic and may be useful for serologic assays. The identification of these antigenic peptides contributes to the understanding of the immunogenicity and persistence of SARS coronavirus. url: https://www.ncbi.nlm.nih.gov/pubmed/14633869/ doi: 10.1373/clinchem.2003.023184 id: cord-299148-uge5uodk author: Wang, Qiang title: A Method To Prevent SARS-CoV-2 IgM False Positives in Gold Immunochromatography and Enzyme-Linked Immunosorbent Assays date: 2020-05-26 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: We set out to investigate the interference factors that led to false-positive novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgM detection results using gold immunochromatography assay (GICA) and enzyme-linked immunosorbent assay (ELISA) and the corresponding solutions. GICA and ELISA were used to detect SARS-CoV-2 IgM in 86 serum samples, including 5 influenza A virus (Flu A) IgM-positive sera, 5 influenza B virus (Flu B) IgM-positive sera, 5 Mycoplasma pneumoniae IgM-positive sera, 5 Legionella pneumophila IgM-positive sera, 6 sera of HIV infection patients, 36 rheumatoid factor IgM (RF-IgM)-positive sera, 5 sera from hypertensive patients, 5 sera from diabetes mellitus patients, and 14 sera from novel coronavirus infection disease 19 (COVID-19) patients. The interference factors causing false-positive reactivity with the two methods were analyzed, and the urea dissociation test was employed to dissociate the SARS-CoV-2 IgM-positive serum using the best dissociation concentration. The two methods detected positive SARS-CoV-2 IgM in 22 mid-to-high-level-RF-IgM-positive sera and 14 sera from COVID-19 patients; the other 50 sera were negative. At a urea dissociation concentration of 6 mol/liter, SARS-CoV-2 IgM results were positive in 1 mid-to-high-level-RF-IgM-positive serum and in 14 COVID-19 patient sera detected using GICA. At a urea dissociation concentration of 4 mol/liter and with affinity index (AI) levels lower than 0.371 set to negative, SARS-CoV-2 IgM results were positive in 3 mid-to-high-level-RF-IgM-positive sera and in 14 COVID-19 patient sera detected using ELISA. The presence of RF-IgM at mid-to-high levels could lead to false-positive reactivity of SARS-CoV-2 IgM detected using GICA and ELISA, and urea dissociation tests would be helpful in reducing SARS-CoV-2 IgM false-positive results. url: https://doi.org/10.1128/jcm.00375-20 doi: 10.1128/jcm.00375-20 id: cord-032689-2xtiiejf author: Wang, Wen-Hung title: A novel, rapid (within hours) culture-free diagnostic method for detecting live Mycobacterium tuberculosis with high sensitivity date: 2020-09-16 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Nucleic acid amplification tests (NAATs) are widely used to diagnose tuberculosis (TB), but cannot discriminate live bacilli from dead bacilli. Live bacilli can be isolated by culture methods, but this is time-consuming. We developed a de novo TB diagnostic method that detects only live bacilli with high sensitivity within hours. METHODS: A prospective study was performed in Taiwan from 2017 to 2018. Sputum was collected consecutively from 1102 patients with suspected TB infection. The sputum was pretreated and heated at 46°C for 1 h to induce the secretion of MPT64 protein from live Mycobacterium tuberculosis. MPT64 was detected with our ultrasensitive enzyme-linked immunosorbent assay (ELISA) coupled with thionicotinamide-adenine dinucleotide (thio-NAD) cycling. We compared our data with those obtained using a culture test (MGIT), a smear test (Kinyoun staining), and a NAAT (Xpert). FINDINGS: The limit of detection for MPT64 in our culture-free ultrasensitive ELISA was 2.0 × 10(−19) moles/assay. When the criterion for a positive response was set as an absorbance value ≥17 mAbs, this value corresponded to ca. 330 CFU/mL in the culture method – almost the same high-detection sensitivity as the culture method. To confirm that MPT64 is secreted from only live bacilli, M. bovis BCG was killed using 8 μg/mL rifampicin and then heated. Following this procedure, our method detected no MPT64. Our rapid ultra-sensitive ELISA-based method required only 5 h to complete. Comparing the results of our method with those of culture tests for 944 specimens revealed a sensitivity of 86.9% (93/107, 95% CI: 79.0–92.7%) and a specificity of 92.0% (770/837, 95% CI: 89.9–93.7%). The performance data were not significantly different (McNemar's test, P = 0.887) from those of the Xpert tests. In addition, at a ≥1+ titer in the smear test, the positive predictive value of our culture-free ultrasensitive ELISA tests was in a good agreement with that of the culture tests. Furthermore, our culture-free ultrasensitive ELISA test had better validity for drug effectiveness examination than Xpert tests because our test detected only live bacilli. INTERPRETATION: Our culture-free ultrasensitive ELISA method detects only live TB bacilli with high sensitivity within hours, allowing for rapid diagnosis of TB and monitoring drug efficacy. FUNDING: Matching Planner Program from JST (VP29117939087), the A-STEP Program from JST (AS3015096U), Waseda University grants for Specific Research Projects (2017A-015 and 2019C-123), the Precise Measurement Technology Promotion Foundation to E.I. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7501073/ doi: 10.1016/j.ebiom.2020.103007 id: cord-004101-0r2g5p1i author: Wang, Yu title: Self-assembly into virus–like particles of the recombinant capsid protein of porcine circovirus type 3 and its application on antibodies detection date: 2020-01-07 words: 4241.0 sentences: 232.0 pages: flesch: 58.0 cache: ./cache/cord-004101-0r2g5p1i.txt txt: ./txt/cord-004101-0r2g5p1i.txt summary: The purified eCap and dCap were identified by transmission electron microscopy (TEM), a large number of round hollow particles with a diameter of 10 nm virus-like particles (VLPs) were observed in eCap, whereas irregular aggregation of proteins observed in dCap. After formation the VLPs were applied as a coating antigen to establish an indirect ELISA (I-ELISA) for detection of PCV3-specific antibody in swine serum. To the best of our knowledge, this is the first report of self-assembly into VLPs of PCV3 recombinant Cap. Our results demonstrated that the VLP-based I-ELISA will be a valuable tool for detecting the presence of PCV3 antibodies in serum samples and will facilitate screening of large numbers of swine serum for clinical purposes. Generation in yeast of recombinant virus-like particles of porcine circovirus type 2 capsid protein and their use for a serologic assay and development of monoclonal antibodies abstract: PCV3 capsid protein (Cap) is an important antigen for diagnosis and vaccine development. To achieve high-level expression of recombinant PCV3 Cap in Escherichia coli (E. coli), the gene of wild-type entire Cap (wt-eCap) was amplified from clinical samples, and three optimized entire Cap (opti-eCap) and one optimized Cap deleted nuclear location signal (NLS) (opti-dCap) gene fragments encoding the same amino acid sequence with wt-eCap were synthesized based on the codon bias of E. coli. Those gene fragments were inserted into the pET30a expression vector. One recombinant strain with the highest expressed soluble eCap from four entire Cap (one wt-eCap and three opti-eCap) and one recombinant strain expressed opti-dCap were selected for further purification. The purified eCap and dCap were identified by transmission electron microscopy (TEM), a large number of round hollow particles with a diameter of 10 nm virus-like particles (VLPs) were observed in eCap, whereas irregular aggregation of proteins observed in dCap. After formation the VLPs were applied as a coating antigen to establish an indirect ELISA (I-ELISA) for detection of PCV3-specific antibody in swine serum. 373 clinical swine serum samples from China collected in 2019 were tested utilizing the VLP-based I-ELISA method under optimized conditions. To the best of our knowledge, this is the first report of self-assembly into VLPs of PCV3 recombinant Cap. Our results demonstrated that the VLP-based I-ELISA will be a valuable tool for detecting the presence of PCV3 antibodies in serum samples and will facilitate screening of large numbers of swine serum for clinical purposes. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6946787/ doi: 10.1186/s13568-019-0940-0 id: cord-305516-s1lvhknm author: Watanabe, Shumpei title: Epizootology and experimental infection of Yokose virus in bats date: 2008-09-11 words: 4133.0 sentences: 232.0 pages: flesch: 55.0 cache: ./cache/cord-305516-s1lvhknm.txt txt: ./txt/cord-305516-s1lvhknm.txt summary: To reveal whether bats serve as an amplifying host for Yokose virus (YOKV), we conducted a serological survey and experimentally infected fruit bats with YOKV isolated from microbats in Japan. To detect antibodies against YOKV, we developed an enzyme-linked immunosorbent assay (ELISA) using biotinylated anti-bat IgG rabbit sera. To detect antibodies against YOKV, we developed an ELISA using biotinylated anti-bat IgG rabbit sera. Using the conventional ELISA, a serological survey was performed on bat serum samples collected from the Philippines and Malaysia. To obtain sera positive for anti-YOKV antibodies, two bats were immunized with inactivated and purified virus. Furthermore, the serum samples collected from bats which were obtained from Japan Zoos were also screened by ELISA using the JEV antigen. We developed an ELISA system using biotin-labeled anti-bat-IgG rabbit serum to detect antibodies against YOKV in bat sera and conducted serological surveys using this system. abstract: To reveal whether bats serve as an amplifying host for Yokose virus (YOKV), we conducted a serological survey and experimentally infected fruit bats with YOKV isolated from microbats in Japan. YOKV belongs to the Entebbe bat virus group of vector unknown group within the genus Flavivirus and family Flaviviridae. To detect antibodies against YOKV, we developed an enzyme-linked immunosorbent assay (ELISA) using biotinylated anti-bat IgG rabbit sera. Serological surveillance was conducted with samples collected in the Philippines and the sera supplied from Malaysia. One of the 36 samples from the Philippines (2.7%) and 5 of the 26 samples from Malaysia (19%) had detectable ELISA antibodies. In the experimental infections, no clinical signs of disease were observed. Moreover, no significant viral genome amplification was detected. These findings revealed that YOKV replicates poorly in the fruit bat, suggesting that fruit bats do not seem to serve as an amplifying host for YOKV. url: https://api.elsevier.com/content/article/pii/S0147957108000428 doi: 10.1016/j.cimid.2008.07.008 id: cord-007495-gpz4gkv3 author: Weiss, M. title: Antibodies to berne virus in horses and other animals date: 2002-11-13 words: 2113.0 sentences: 103.0 pages: flesch: 51.0 cache: ./cache/cord-007495-gpz4gkv3.txt txt: ./txt/cord-007495-gpz4gkv3.txt summary: Positive reactions were also obtained in serum neutralization tests and ELISA using small numbers of horse sera from Germany, France and the U.S.A. The results of neutralization tests and ELISA were correlated in 83% of random samples tested; 13% were neutralization-positive and ELISA-negative and in 4% the inverse was observed. The activity of Berne virus in the Swiss adult horse population was evalPositive reactions in serum neutralization of small numbers of randomlycollected equine sera from Germany (8/11) and the USA (24/38) and in ELISA of samples from Southern France (10/28) and the USA (12/16) were recorded. As shown in Table IV , high percentages of serum samples with elevated neutralization titers were encountered in all ungulates studied (horse, cattle, goat, sheep and pig). We have preliminary evidence (from neutralization tests using 31 randomly collected cattle sera) that a Berne-related virus is active also in the Dutch cattle population; in only 3 samples titers were <10 and in 10 sera values of >50 were recorded. abstract: After inoculation into 2 foals, Berne virus induced neutralizing antibody, but did not cause clinical symptoms. In a horizontal study of seropositive mares and their offspring, a decline of maternal antibodies and a sudden synchronous seroconversion in all foals were observed, again without clinical symptoms. The virus is widespread in the Swiss horse population and has been so during the last decade; rises in antibody titers were noted in 9% of paired sera sampled at random. Positive reactions were also obtained in serum neutralization tests and ELISA using small numbers of horse sera from Germany, France and the U.S.A. The results of neutralization tests and ELISA were correlated in 83% of random samples tested; 13% were neutralization-positive and ELISA-negative and in 4% the inverse was observed. Neutralizing activity was found in the sera of other ungulates (cattle, goat, sheep and pig), laboratory rabbits and 2 species of wild mice (Clethrionomys glareolus and Apodemus sylvaticus). Inconclusive results were obtained with feline and human sera; those from dogs and foxes (Vulpes vulpes) were consistently negative. The probable occurrence of antigenic variants in Berne-type viruses is discussed. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117441/ doi: 10.1016/0378-1135(84)90014-2 id: cord-003656-7mzsaz7a author: Wium, Martha title: DNA Vaccines Against Mycoplasma Elicit Humoral Immune Responses in Ostriches date: 2019-05-14 words: 5575.0 sentences: 308.0 pages: flesch: 53.0 cache: ./cache/cord-003656-7mzsaz7a.txt txt: ./txt/cord-003656-7mzsaz7a.txt summary: Currently there are no commercially available vaccines against ostrich-infecting mycoplasmas and this study therefore set out to develop and evaluate the use of a DNA vaccine against mycoplasma infections in ostriches using an OppA protein as antigen. The ability of the DNA vaccines to elicit an anti-OppA antibody response was evaluated by ELISA using the recombinant OppA protein of Ms03 as coating antigen. In this study we report, for the first time, that a DNA vaccine can elicit a humoral immune response in ostriches using OppA as antigen. The controls were serum samples representing the week 0, 3, 6, and 9 sampling points of a single ostrich, randomly selected from the pCI-neo_oppA 1,200 µg group based on high titers produced after vaccination. In this study, DNA vaccines were developed for ostriches using the oppA gene of an ostrich-infecting mycoplasma (Ms03) as vaccine antigen. abstract: In ostriches, the population densities resulting from intensive rearing increases susceptibility to pathogens such as mycoplasmas. In addition to good management practices, vaccination offers an attractive alternative for controlling mycoplasma infections in food animals, instead of using antibiotics, which often leave unacceptable residues. The use of live attenuated vaccines, however, carry the concern of reversion to virulence or genetic recombination with field strains. Currently there are no commercially available vaccines against ostrich-infecting mycoplasmas and this study therefore set out to develop and evaluate the use of a DNA vaccine against mycoplasma infections in ostriches using an OppA protein as antigen. To this end, the oppA gene of “Mycoplasma nasistruthionis sp. nov.” str. Ms03 was cloned into two DNA vaccine expression vectors after codon correction by site-directed mutagenesis. Three-months-old ostriches were then vaccinated intramuscularly at different doses followed by a booster vaccination after 6 weeks. The ability of the DNA vaccines to elicit an anti-OppA antibody response was evaluated by ELISA using the recombinant OppA protein of Ms03 as coating antigen. A statistically significant anti-OppA antibody response could be detected after administration of a booster vaccination indicating that the OppA protein was successfully immunogenic. The responses were also both dose and vector dependent. In conclusion, the DNA vaccines were able to elicit an immune response in ostriches and can therefore be viewed as an option for the development of vaccines against mycoplasma infections. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6527592/ doi: 10.3389/fimmu.2019.01061 id: cord-264968-ctx39vhi author: Woo, Patrick CY title: Relative rates of non-pneumonic SARS coronavirus infection and SARS coronavirus pneumonia date: 2004-03-13 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Although the genome of severe acute respiratory syndrome coronavirus (SARS-CoV) has been sequenced and a possible animal reservoir identified, seroprevalence studies and mass screening for detection of subclinical and non-pneumonic infections are still lacking. METHODS: We cloned and purified the nucleocapsid protein and spike polypeptide of SARS-CoV and examined their immunogenicity with serum from patients with SARS-CoV pneumonia. An ELISA based on recombinant nucleocapsid protein for IgG detection was tested with serum from 149 healthy blood donors who donated 3 years previously and with serum positive for antibodies against SARS-CoV (by indirect immunofluorescence assay) from 106 patients with SARS-CoV pneumonia. The seroprevalence of SARS-CoV was studied with the ELISA in healthy blood donors who donated during the SARS outbreak in Hong Kong, non-pneumonic hospital inpatients, and symptom-free health-care workers. All positive samples were confirmed by two separate western-blot assays (with recombinant nucleocapsid protein and recombinant spike polypeptide). FINDINGS: Western-blot analysis showed that the nucleocapsid protein and spike polypeptide of SARS-CoV are highly immunogenic. The specificity of the IgG antibody test (ELISA with positive samples confirmed by the two western-blot assays) was 100%, and the sensitivity was 94·3%. Three of 400 healthy blood donors who donated during the SARS outbreak and one of 131 non-pneumonic paediatric inpatients were positive for IgG antibodies, confirmed by the two western-blot assays (total, 0·48% of our study population). INTERPRETATION: Our findings support the existence of subclinical or non-pneumonic SARS-CoV infections. Such infections are more common than SARS-CoV pneumonia in our locality. url: https://www.sciencedirect.com/science/article/pii/S0140673604157292 doi: 10.1016/s0140-6736(04)15729-2 id: cord-344871-486sk4wc author: Wu, Jianping title: Biochemical and structural characterization of the interface mediating interaction between the influenza A virus non-structural protein-1 and a monoclonal antibody date: 2016-09-16 words: 6992.0 sentences: 379.0 pages: flesch: 58.0 cache: ./cache/cord-344871-486sk4wc.txt txt: ./txt/cord-344871-486sk4wc.txt summary: We have previously shown that a non-structural protein 1 (NS1)-binding monoclonal antibody, termed as 2H6, can significantly reduce influenza A virus (IAV) replication when expressed intracellularly. As comparative ELISA in this and previous studies 29 showed that residues N48 and T49 in NS1(RBD) are important for the interaction with mAb 2H6, they were defined as active residues involved in the binding interaction to generate a series of models of the NS1(RBD) and 2H6-Fab complex. Overall, the predicted model from cluster 2 is consistent with our comparative ELISA data and suggests that residues N48 and T49 are important for the binding between NS1(RBD) and 2H6-Fab because their side-chains could make hydrogen bonds with residues in the VH-CDR2 of the Fab. In addition, R44 of NS1(RBD) was distal from the antibody-antigen interface, which is consistent with the results from comparative ELISA ( Figure S1 ) showing that substitution of R44 of NS1(RBD) with K did not affect its interaction with mAb 2H6. abstract: We have previously shown that a non-structural protein 1 (NS1)-binding monoclonal antibody, termed as 2H6, can significantly reduce influenza A virus (IAV) replication when expressed intracellularly. In this study, we further showed that 2H6 binds stronger to the NS1 of H5N1 than A/Puerto Rico/8/1934(H1N1) because of an amino acid difference at residue 48. A crystal structure of 2H6 fragment antigen-binding (Fab) has also been solved and docked onto the NS1 structure to reveal the contacts between specific residues at the interface of antibody-antigen complex. In one of the models, the predicted molecular contacts between residues in NS1 and 2H6-Fab correlate well with biochemical results. Taken together, residues N48 and T49 in H5N1 NS1 act cooperatively to maintain a strong interaction with mAb 2H6 by forming hydrogen bonds with residues found in the heavy chain of the antibody. Interestingly, the pandemic H1N1-2009 and the majority of seasonal H3N2 circulating in humans since 1968 has N48 in NS1, suggesting that mAb 2H6 could bind to most of the currently circulating seasonal influenza A virus strains. Consistent with the involvement of residue T49, which is well-conserved, in RNA binding, mAb 2H6 was also found to inhibit the interaction between NS1 and double-stranded RNA. url: https://doi.org/10.1038/srep33382 doi: 10.1038/srep33382 id: cord-275793-k0uvqcmp author: Xia, Hongyan title: A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies date: 2010-04-18 words: 2648.0 sentences: 154.0 pages: flesch: 53.0 cache: ./cache/cord-275793-k0uvqcmp.txt txt: ./txt/cord-275793-k0uvqcmp.txt summary: title: A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies This study describes a novel blocking microsphere-based immunoassay for highly sensitive and specific detection of antibodies against bovine viral diarrhoea virus (BVDV). The objectives of this study were to develop a blocking microsphere-based immunoassay (bMIA) for detection of antibodies against BVDV, and to compare the performance of the assay with a commercial ELISA kit. This study describes the development and evaluation of a microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies. The diagnostic performance of the new bMIA was compared to that of a commercial blocking ELISA system, by testing a large panel of 509 bovine sera. A simple, rapid and reliable enzyme-linked immunosorbent assay for the detection of bovine virus diarrhoea virus (BVDV) specific antibodies in cattle serum, plasma and bulk milk abstract: This study describes a novel blocking microsphere-based immunoassay for highly sensitive and specific detection of antibodies against bovine viral diarrhoea virus (BVDV). The intra- and inter-assay variability are 4.9% and less than 7%, respectively, and variability of bead conjugations is less than 6.6%. The diagnostic performance of the assay was evaluated by testing a total of 509 serum samples. Based on a negative/positive cut-off value of 30.3%, the assay has a sensitivity of 99.4% and a specificity of 98.3% relative to ELISA. The new microsphere immunoassay provides an alternative to conventional ELISA systems and can be used for high-throughput screening in the BVD control programmes. url: https://doi.org/10.1016/j.jviromet.2010.04.009 doi: 10.1016/j.jviromet.2010.04.009 id: cord-003208-lwirkob3 author: Yan, Liping title: Novel protein chip for the detection of antibodies against infectious bronchitis virus date: 2018-09-17 words: 3973.0 sentences: 220.0 pages: flesch: 52.0 cache: ./cache/cord-003208-lwirkob3.txt txt: ./txt/cord-003208-lwirkob3.txt summary: RESULTS: We have developed two indirect microarray methods to detect antibodies against IBV: a chemiluminescent immunoassay test (CIT) and a rapid diagnostic test (RDT). Compared with these methods, enzyme-linked immunosorbent assay (ELISA) has been widely used for testing IBV early infection and continuous infection, and this technique can be used for both antigenic and antibody detection. The data showed that 130 serum samples were positive for antibodies against IBV, and 14 samples were negative, similar to the results of the IDEXX IBV Ab Test kit with the nsp5 concentration of 0.2 mg/mL (Table 4 ). The specific experiments of the RDT showed that no cross-reaction Fig. 4 a Distribution of the SNRs of the IDEXX-positive (n = 142) and IDEXX-negative (n = 42) serum samples of the clinical sera obtained from the IBV protein microarray. abstract: BACKGROUND: Infectious bronchitis (IB) caused by the IB virus (IBV) can cause acute damage to chickens around the world. Therefore, rapid diagnosis and immune status determination are critical for controlling IBV outbreaks. Enzyme-linked immunosorbent assays (ELISAs) have been widely used in the detection of IBV antibodies in the early infection and continuous infection of IB because they are more sensitive and quicker than other diagnostic methods. RESULTS: We have developed two indirect microarray methods to detect antibodies against IBV: a chemiluminescent immunoassay test (CIT) and a rapid diagnostic test (RDT). IBV nonstructural protein 5 (nsp5) was expressed, purified from Escherichia coli, and used to spot the initiator integrated poly(dimethylsiloxane), which can provide a near “zero” background for serological assays. Compared with the IDEXX IBV Ab Test kit, CIT and RDT have a sensitivity and specificity of at least 98.88% and 91.67%, respectively. No cross-reaction was detected with antibodies against avian influenza virus subtypes (H5, H7, and H9), Newcastle disease virus, Marek’s disease virus, infectious bursal disease virus, and chicken anemia virus. The coefficients of variation of the reproducibility of the intra- and inter-assays for CIT ranged from 0.8 to 18.63%. The reproducibility of RDT was consistent with the original results. The application of the IBV nsp5 protein microarray showed that the positive rate of the CIT was 96.77%, that of the nsp5 ELISA was 91.40%, and that of the RDT was 90.32%. Furthermore, the RDT, which was visible to the naked eye, could be completed within 15 min. Our results indicated that compared with nsp5 ELISA, the CIT was more sensitive, and the RDT had similar positive rates but was faster. Furthermore, the two proposed methods were specific and stable. CONCLUSIONS: Two microarray assays, which were rapid, specific, sensitive, and relatively simple, were developed for the detection of an antibody against IBV. These methods can be of great value for the surveillance of pathogens and monitoring the efficiency of vaccination. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6142349/ doi: 10.1186/s12917-018-1586-x id: cord-348660-qnbgywgy author: Yilmaz, Huseyin title: Production of Recombinant N Protein of Infectious Bronchitis Virus Using the Baculovirus Expression System and Its Assessment as a Diagnostic Antigen date: 2018-07-09 words: 3921.0 sentences: 183.0 pages: flesch: 43.0 cache: ./cache/cord-348660-qnbgywgy.txt txt: ./txt/cord-348660-qnbgywgy.txt summary: Following optimization of the ELISA protocol, 18 test sera were obtained from broiler chickens exposed to natural wild-type IBV infection, 18 test sera from broiler chickens vaccinated with a live-attenuated commercial IBV vaccine, and sera obtained at different time-points from chicks immunized with recombinant IBV N protein (described above) were analyzed to detect IBV N-specific antibodies. To assess the reliability of the performance of our in-house indirect IBV N ELISA, a panel of sera was obtained from chickens naturally infected with local wild-type IBV strains, chickens vaccinated with live-attenuated commercial IBV vaccine, and chickens immunized with recombinant IBV N protein (expressed in a baculovirus expression system). This represents the first study in Turkey that expressed recombinant IBV N protein in baculovirus and examined its reactivity against antisera obtained from Turkish chickens for potential use as antigen Fig. 5 Detection of IBV N specific antibodies in sera obtained from naturally infected chicken (a) and vaccinated chickens (b) using an in-house IBV-N ELISA and a commercial ELISA. abstract: The avian coronavirus-infectious bronchitis virus (AvCoV-IBV) is recognized as an important avian pathogen, and new viral variants are a continuous threat to the poultry industry worldwide. Sensitive diagnostics and efficacious vaccines are necessary to combat IBV infections in chickens. The aim of this study was to produce recombinant N protein of IBV in the baculovirus system to use in ELISA diagnostic tests in order to enable the assessment of the sero-prevalence and risk of IBV infections in chickens in Turkey. For this, the gene encoding the N protein of the Beaudette strain of IBV was expressed using a recombinant baculovirus expression system. The recombinant N protein was purified using Ni-NTA affinity chromatography. An estimated 50-kDa recombinant protein corresponding to the expected molecular weight of IBV N including the 6xHis tag was detected using an anti-His monoclonal antibody. Specific immunoreactivity of the recombinant protein was confirmed by Western blot using antiserum obtained from vaccinated and naturally infected chicken from Turkey as well as using a monoclonal antibody raised against the N protein of the IBV Massachusetts strain. The results obtained with the in-house ELISA had high agreement with a commercial ELISA. Immunoreactivity analysis using antisera in Western blotting and the in-house ELISA suggests that the recombinant IBV N protein could be broadly cross-reactive with antisera produced against different IBV strains. We conclude that the recombinant baculovirus expressed IBV N protein could serve as a useful diagnostic antigen for detection of IBV infections in chickens by ELISA. url: https://www.ncbi.nlm.nih.gov/pubmed/29987628/ doi: 10.1007/s12010-018-2815-2 id: cord-300908-i80tuhqk author: Yu, Fuxun title: Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA date: 2015-08-04 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe and affordable serodiagnostic methods for proper clinical management of infected patients. METHODS: The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system. RESULTS: The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively. CONCLUSIONS: The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations. url: https://www.ncbi.nlm.nih.gov/pubmed/26239826/ doi: 10.1186/s12985-015-0350-0 id: cord-251943-jzaeaxam author: Zhang, Jian‐San title: A serological survey on neutralizing antibody titer of SARS convalescent sera date: 2005-08-24 words: 2542.0 sentences: 139.0 pages: flesch: 50.0 cache: ./cache/cord-251943-jzaeaxam.txt txt: ./txt/cord-251943-jzaeaxam.txt summary: A seroepidemiologic study was conducted in North China in 2003 to determine the neutralizing antibody titer of severe acute respiratory syndrome (SARS) convalescent sera. A total of 99 SARS convalescent serum samples were collected from patients from the Inner Mongolia Autonomous Region, Hebei Province, and Beijing 35–180 days after the onset of symptoms. To gain a comprehensive understanding of the antibody to SARS-CoV, we report the anti-SARS antibody titer of 87 SARS convalescent sera determined by neutralization assay. These 87 serum samples were confirmed to be positive for anti-SARS antibodies with the combination of ELISA, neutralization, and Western blot, so they were pooled to form a convalescent sera database for the further analysis of neutralizing antibody titer. The anti-SARS neutralizing antibody titer of 87 positive convalescent sera was analyzed quantitatively by the neutralization assay. In our laboratory, a combination of ELISA, neutralization assay, and Western blot were performed on 99 SARS convalescent sera. abstract: A seroepidemiologic study was conducted in North China in 2003 to determine the neutralizing antibody titer of severe acute respiratory syndrome (SARS) convalescent sera. A total of 99 SARS convalescent serum samples were collected from patients from the Inner Mongolia Autonomous Region, Hebei Province, and Beijing 35–180 days after the onset of symptoms. The anti‐SARS antibodies were detected by enzyme‐linked immunosorbent assay (ELISA), neutralization assay, and Western blot. Eighty‐seven serum samples were confirmed to be positive for SARS antibodies. The neutralizing antibody titer of the 87 positive sera was analyzed quantitatively by neutralization assay. The geometric mean titer (GMT) of the 87 convalescent sera was 1:61. The Kolmogorov–Smirnov test showed that the neutralizing antibody titers conform to normal distribution, which suggests that the average anti‐SARS antibody level in this study was representative of the convalescent antibody level of the SARS population. This result could be useful for the development and quality control of SARS vaccines. J. Med. Virol. 77:147–150, 2005. © 2005 Wiley‐Liss, Inc. url: https://www.ncbi.nlm.nih.gov/pubmed/16121363/ doi: 10.1002/jmv.20431 id: cord-316723-srenbxa7 author: Zhao, Jincun title: Development and evaluation of an enzyme-linked immunosorbent assay for detection of antibodies against the spike protein of SARS-coronavirus date: 2004-11-23 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by infection with SARS-associated coronavirus (CoV). Amino acid residues 450–650 of the spike (S) glycoprotein of SARS-CoV (S450-650) contains dominant epitopes for anti-viral antibodies (Abs) in patient sera. OBJECTIVES: To develop and evaluate an ELISA system for detection of anti-S Abs in patient sera. STUDY DESIGN: Express recombinant S450-650 in E. Coli and evaluate the sensitivity and specificity of an ELISA system based on the S450-650 polypeptide. RESULTS: The S450-650-based ELISA detected IgG Abs in 41 out of 51 serum samples from 22 hospitalized patients with probable SARS, a result closely correlated with that obtained with a virus-based ELISA (r = 0.75, k = 0.8). Differential anti-S IgG responses were observed amongst SARS patients. Some of them produced anti-S Abs early during their infection, while others failed to make IgG Abs against the S450-650 polypeptide. None of the serum samples from 100 healthy blood donors was positive in the S450-650-based assay. CONCLUSION: The S450-650-based ELISA can detect anti-S IgG Abs with high sensitivity and specificity. url: https://www.ncbi.nlm.nih.gov/pubmed/15797360/ doi: 10.1016/j.jcv.2004.09.024 id: cord-000434-ff2zadol author: Zhao, Rongmao title: Identification of a Highly Conserved H1 Subtype-Specific Epitope with Diagnostic Potential in the Hemagglutinin Protein of Influenza A Virus date: 2011-08-19 words: 5752.0 sentences: 307.0 pages: flesch: 51.0 cache: ./cache/cord-000434-ff2zadol.txt txt: ./txt/cord-000434-ff2zadol.txt summary: The highly conserved H1 subtype-specific immunodominant epitope may form the basis for developing novel assays for sero-diagnosis and active surveillance against H1N1 IAVs. Influenza A viruses (IAVs), members of the Orthomyxoviridae family, are highly contagious to a variety of avian and mammalian species. To confirm that these antibodies can recognize the HA antigen, the reactivity of the anti-peptide sera were evaluated by Western blot and ELISA against the purified HA0 protein of H1N1pdm virus. The sensitivity and specificity of peptide-ELISA versus HI test was 96.5% and 74.4%, respectively, indicating the potential of the peptide-ELISA method in detecting antibody against H1-subtype IAVs. In the present study, we identified immunodominant linear B cell epitopes on the H1N1pdm virus HA protein by a peptide scanning approach using H1N1pdm patients sera. To screen the H1-subtype specific epitopes, a set of 50 peptides spanning the amino acid sequences of the HA protein ectodomain of pandemic A/H1N1 2009 (H1N1pdm) influenza virus strain A/ California/04/2009 were synthesized. abstract: Subtype specificity of influenza A virus (IAV) is determined by its two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). For HA, 16 distinct subtypes (H1–H16) exist, while nine exist for NA. The epidemic strains of H1N1 IAV change frequently and cause annual seasonal epidemics as well as occasional pandemics, such as the notorious 1918 influenza pandemic. The recent introduction of pandemic A/H1N1 IAV (H1N1pdm virus) into humans re-emphasizes the public health concern about H1N1 IAV. Several studies have identified conserved epitopes within specific HA subtypes that can be used for diagnostics. However, immune specific epitopes in H1N1 IAV have not been completely assessed. In this study, linear epitopes on the H1N1pdm viral HA protein were identified by peptide scanning using libraries of overlapping peptides against convalescent sera from H1N1pdm patients. One epitope, P5 (aa 58–72) was found to be immunodominant in patients and to evoke high titer antibodies in mice. Multiple sequence alignments and in silico coverage analysis showed that this epitope is highly conserved in influenza H1 HA [with a coverage of 91.6% (9,860/10,767)] and almost completely absent in other subtypes [with a coverage of 3.3% (792/23,895)]. This previously unidentified linear epitope is located outside the five well-recognized antigenic sites in HA. A peptide ELISA method based on this epitope was developed and showed high correlation (χ(2) = 51.81, P<0.01, Pearson correlation coefficient R = 0.741) with a hemagglutination inhibition test. The highly conserved H1 subtype-specific immunodominant epitope may form the basis for developing novel assays for sero-diagnosis and active surveillance against H1N1 IAVs. url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3158760/ doi: 10.1371/journal.pone.0023374 id: cord-307110-eiobmxp2 author: Zhao, Shan title: Serological Screening for Coronavirus Infections in Cats date: 2019-08-13 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: Coronaviruses (CoVs) are widespread among mammals and birds and known for their potential for cross-species transmission. In cats, infections with feline coronaviruses (FCoVs) are common. Several non-feline coronaviruses have been reported to infect feline cells as well as cats after experimental infection, supported by their ability to engage the feline receptor ortholog for cell entry. However, whether cats might become naturally infected with CoVs of other species is unknown. We analyzed coronavirus infections in cats by serological monitoring. In total 137 cat serum samples and 25 FCoV type 1 or type 2-specific antisera were screened for the presence of antibodies against the S1 receptor binding subunit of the CoV spike protein, which is immunogenic and possesses low amino acid sequence identity among coronavirus species. Seventy-eight sera were positive for antibodies that recognized one or more coronavirus S1s whereas 1 serum exclusively reacted with human coronavirus 229E (HCoV-229E) and two sera exclusively reacted with porcine delta coronavirus (PDCoV). We observed antigenic cross-reactivity between S1s of type 1 and type 2 FCoVs, and between FCoV type 1 and porcine epidemic diarrhea virus (PEDV). Domain mapping of antibody epitopes indicated the presence of conserved epitope(s) particularly in the CD domains of S1. The cross-reactivity of FCoV type 1 and PEDV was also observed at the level of virus neutralization. To conclude, we provide the first evidence of antigenic cross-reactivity among S1 proteins of coronaviruses, which should be considered in the development of serological diagnoses. In addition, the potential role of cats in cross-species transmission of coronaviruses cannot be excluded. url: https://www.ncbi.nlm.nih.gov/pubmed/31412572/ doi: 10.3390/v11080743 id: cord-317026-9zgc6xrb author: Zhao, Shan title: Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus date: 2019-11-30 words: 6223.0 sentences: 317.0 pages: flesch: 55.0 cache: ./cache/cord-317026-9zgc6xrb.txt txt: ./txt/cord-317026-9zgc6xrb.txt summary: In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure. On the other hand, serological assays can be used to support the diagnosis of a clinical ECoV infection by showing seroconversion or a significant increase in antibody titer in paired serum samples. For the six horses that did not show a significant rise in VNT, five serum samples collected at the acute stage already had high antibody levels as shown by ELISA and neutralization assay (mean OD value > 2.6, mean VNT > 9, Table S1 ). For the six horses that did not show a significant rise in VNT, five serum samples collected at the acute stage already had high antibody levels as shown by ELISA and neutralization assay (mean OD value > 2.6, mean VNT > 9, Table S1 ). abstract: Equine coronavirus (ECoV) is considered to be involved in enteric diseases in foals. Recently, several outbreaks of ECoV infection have also been reported in adult horses from the USA, France and Japan. Epidemiological studies of ECoV infection are still limited, and the seroprevalence of ECoV infection in Europe is unknown. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure. Both formats showed high diagnostic accuracy compared to virus neutralization (VN) assay. Receiver-operating characteristic (ROC) analyses were performed to determine the best cut-off values for both ELISA formats, assuming a test specificity of 99%. Employing the developed ELISA method, we detected seroconversion in 70.4% of horses from an ECoV outbreak. Among the 1084 horse sera, seropositivity varied from 25.9% (young horses) to 82.8% (adult horses) in Dutch horse populations. Further, sera of Icelandic horses were included in this study and a significant number of sera (62%) were found to be positive. Overall, the results demonstrated that the ECoV S1-based ELISA has reliable diagnostic performance compared to the VN assay and is a useful assay to support seroconversion in horses involved with ECoV outbreaks and to estimate ECoV seroprevalence in populations of horses. url: https://www.ncbi.nlm.nih.gov/pubmed/31801275/ doi: 10.3390/v11121109 id: cord-299189-59d4aojh author: Zou, Hao title: Transmissible gastroenteritis virus: Identification of M protein-binding peptide ligands with antiviral and diagnostic potential date: 2013-07-02 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: The membrane (M) protein is one of the major structural proteins of coronavirus particles. In this study, the M protein of transmissible gastroenteritis virus (TGEV) was used to biopan a 12-mer phage display random peptide library. Three phages expressing TGEV-M-binding peptides were identified and characterized in more depth. A phage-based immunosorbent assay (phage-ELISA) capable of differentiating TGEV from other coronaviruses was developed using one phage, phTGEV-M7, as antigen. When the phage-ELISA was compared to conventional antibody-based ELISA for detecting infections, phage-ELISA exhibited greater sensitivity. A chemically synthesized, TGEV-M7 peptide (pepTGEV-M7; HALTPIKYIPPG) was evaluated for antiviral activity. Plaque-reduction assays revealed that pepTGEV-M7 was able to prevent TGEV infection in vitro (p < 0.01) following pretreatment of the virus with the peptide. Indirect immunofluorescence and real-time RT-PCR confirmed the inhibitory effects of the peptide. These results indicate that pepTGEV-M7 might be utilized for virus-specific diagnostics and treatment. url: https://www.sciencedirect.com/science/article/pii/S0166354213001721 doi: 10.1016/j.antiviral.2013.06.015 id: cord-001521-l36f1gp7 author: nan title: Oral and Poster Manuscripts date: 2011-04-08 words: 183363.0 sentences: 11362.0 pages: flesch: 53.0 cache: ./cache/cord-001521-l36f1gp7.txt txt: ./txt/cord-001521-l36f1gp7.txt summary: The IC 50 values determined in functional NI assays provide valuable information for detection of resistant viruses, but should not be used to draw direct correlations with drug concentrations needed to inhibit virus replication in the infected human host, as clinical data to support such inferences are inadequate. • Standardized reagents and protocols • Choice of detection technology • Simple instrumentation requirements • High sensitivity for use with low virus concentrations • Compatibility with batch-mode processing and largescale assay throughput • Broad specificity of influenza detection • Flexibility in assay format • Additional NA assay applications -cell-based viral assays, screening for new NIs, detection of NA from other organisms Functional neuraminidase inhibition assays enable detection of any resistance mutation and are extremely important in conjunction with sequence-based screening assays for global monitoring of virus isolates for NI resistance mutations, including known and new mutations. Such new assays need to include methods to measure local antibodies and virus-specific lymphocytes, especially in the case of live attenuated influenza vaccines, because of their potential to induce such broad-based immune responses. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4313891/ doi: 10.1111/j.1750-2659.2011.00209.x id: cord-006828-i88on326 author: nan title: Abstracts DGRh-Kongress 2013 date: 2013-09-15 words: 30772.0 sentences: 2576.0 pages: flesch: 52.0 cache: ./cache/cord-006828-i88on326.txt txt: ./txt/cord-006828-i88on326.txt summary: Comparing gene expression profiles of yellow fever immunized individuals and active SLE patients it was possible to identify a "common" and an "autoimmune-specific" IFN signature. The inflammatory and profibrotic effects upon Aab stimulation in vitro, and their associations with clinical findings suggest a role for autoantibody-mediated activation of immune cells mediated through the AT1R and ETAR in the pathogenesis or even the onset of the disease. This study was aimed to investigate the humoral and cellular immune response to VZV including assessment of IgG-anti-VZV avidity and VZV-specific reactivity of lymphocytes in RA (n=56) or JIA patients (n=75) on different treatments, including biologic agents, such as anti-tumor-necrosis-factor(TNF)-alpha or anti-interleukin-6 (IL-6) receptor inhibition (tocilizumab), compared to 37 healthy adults (HA) and 41 children (HC). Production of cytokines by B cells in response to TLR9 stimulation inversely correlates with disease activity in SLE-patients abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103148/ doi: 10.1007/s00393-013-1255-1 id: cord-006860-a3b8hyyr author: nan title: 40th Annual Meeting of the GTH (Gesellschaft für Thrombose- und Hämostaseforschung) date: 1996 words: 90660.0 sentences: 5152.0 pages: flesch: 50.0 cache: ./cache/cord-006860-a3b8hyyr.txt txt: ./txt/cord-006860-a3b8hyyr.txt summary: Dept of Pediatrics, University Hospitals Kiel and Mtinster, Germany Resistance to activated protein C (APCR), in the majority of cases associated with the Arg 506 Gin point mutation in the factor V gene is present in more than 50 % of patients < 60 years of age with unexplained thrombophilia. The regular APC resistance test is not applicable to plasma from Orally anticoagulated (OAC) or heparinized patients due to decreased levels of vitamin K-dependent clotting factors and to thrombin inhibition by antithrombin, respectively. On admission an extensive coagulation screen yielded the following results (n/normal, t/elevated, I/reduced, +/positive, -/negative): PT t, aPTT t, Tr n, factor II, V, VIII n, factor VII, IX, XI, XII /,, fibrinogan t, ATIII n, protein C, S *, activated protein C sensitivity ratio 1.92 ($), FV-Leidenmutation PCR -, fibrinolytic system n, TAT t, Ft÷2 t, lupus anticoagulant +, heparin induced platelet antibodies +; no diagnosis of a specific autoimmuna disorder could be made. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103196/ doi: 10.1007/bf00641048 id: cord-014462-11ggaqf1 author: nan title: Abstracts of the Papers Presented in the XIX National Conference of Indian Virological Society, “Recent Trends in Viral Disease Problems and Management”, on 18–20 March, 2010, at S.V. University, Tirupati, Andhra Pradesh date: 2011-04-21 words: 35453.0 sentences: 1711.0 pages: flesch: 49.0 cache: ./cache/cord-014462-11ggaqf1.txt txt: ./txt/cord-014462-11ggaqf1.txt summary: Molecular diagnosis based on reverse transcription (RT)-PCR s.a. one step or nested PCR, nucleic acid sequence based amplification (NASBA), or real time RT-PCR, has gradually replaced the virus isolation method as the new standard for the detection of dengue virus in acute phase serum samples. Non-genetic methods of management of these diseases include quarantine measures, eradication of infected plants and weed hosts, crop rotation, use of certified virus-free seed or planting stock and use of pesticides to control insect vector populations implicated in transmission of viruses. The results of this study indicate that NS1 antigen based ELISA test can be an useful tool to detect the dengue virus infection in patients during the early acute phase of disease since appearance of IgM antibodies usually occur after fifth day of the infection. The studies showed high level of expression in case of constructed vector as compared to infected virus for the specific protein. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3639731/ doi: 10.1007/s13337-011-0027-2 id: cord-014965-efmozngq author: nan title: Infectious diseases other than CMV (1st Section) date: 2001-06-11 words: nan sentences: nan pages: flesch: nan cache: txt: summary: abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7091871/ doi: 10.1038/sj.bmt.1702942 id: cord-015021-pol2qm74 author: nan title: Third International Congress on the Immune Consequences of Trauma, Shock and Sepsis —Mechanisms and Therapeutic Approaches date: 1994 words: 162327.0 sentences: 9379.0 pages: flesch: 50.0 cache: ./cache/cord-015021-pol2qm74.txt txt: ./txt/cord-015021-pol2qm74.txt summary: It is our current understanding that LPS is responsible for many of the pathophysiological events observed during gramnegative infections and that one of the major mechanisms leading to shock and death is the LPS-induced activation of macrophages resulting in the production and release of lipid and peptide mediators, among which tumor necrosis factor seems to be the most important. However plasma IL-6 estimation revealed a statistically significant reduction at 6 hours in tanrine-treated animals compared to glycino and TW controls ( Objective: To evaluate the effects of allogeneic blood transfusion, thermal injury and bacterial garage on interteukin 4 (IL-4), tumor necrosis factor alpha (TNF) production and host mortality and to study if the administration of thymopentth (THY) could affect these events. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7095072/ doi: 10.1007/bf02258437 id: cord-015147-h0o0yqv8 author: nan title: Oral Communications and Posters date: 2014-09-12 words: 73711.0 sentences: 3862.0 pages: flesch: 43.0 cache: ./cache/cord-015147-h0o0yqv8.txt txt: ./txt/cord-015147-h0o0yqv8.txt summary: Cyclooxygenases (COX) catalyze the first step in the synthesis of prostaglandins (PG) from arachidonic acid.COX-1 is constitutively expressed.The COX-2 gene is an immediate early-response gene that is induced by variety of mitogenic and inflammatory stimuli.Levels of COX-2 are increased in both inflamed and malignant tissues.In inflamed tissues, there is both pharmacological and genetic evidence that targeting COX-2 can either improve (e.g., osteoarthritis) or exacerbate symptoms (e.g., inflammatory bowel disease).Multiple lines of evidence suggest that COX-2 plays a significant role in carcinogenesis.The most specific data that support a cause-and effect relationship between COX-2 and tumorigenesis come from genetic studies.Overexpression of COX-2 has been observed to drive tumor formation whereas COX-2 deficiency protects against several tumor types.Selective COX-2 inhibitors protect against the formation and growth of experimental tumors.Moreover, selective COX-2 inhibitors are active in preventing colorectal adenomas in humans.Increased amounts of COX-2-derived PGE2 are found in both inflamed and neoplastic tissues.The fact that PGE2 can stimulate cell proliferation, inhibit apoptosis and induce angiogenesis fits with evidence that induction of COX-2 contributes to both wound healing and tumor growth.Taken together, it seems likely that COX-2 induction contributes to wound healing in response to injury but reduces the threshold for carcinogenesis. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7095932/ doi: 10.1007/bf03353884 id: cord-015394-uj7fe5y6 author: nan title: Scientific Abstracts date: 2008-12-23 words: 242330.0 sentences: 15267.0 pages: flesch: 52.0 cache: ./cache/cord-015394-uj7fe5y6.txt txt: ./txt/cord-015394-uj7fe5y6.txt summary: Studies involving immunohistochemical analysis of normal ovaries have shown that granulosa cells express significantly higher levels of the activator protein-1 (AP-1) transcription factor, cFos compared to theca cells, where cFos expression is virtually absent. Following acute hypoxia (0.5% O2) for one to six hours, RhoA mRNA, total protein and activation (RhoA-GTP) levels were analysed, using semi-quantitative PCRs and western blot, and compared to normoxic non-pregnant human uterine smooth muscle control cells. Since there is an urgent need for non-invasive methods for determination of fetal (F) and placental (P) function, this study was designed to evaluate the genes differently and commonly expressed in P tissue and leukocytes in maternal (M) and F circulation.Material and Methods. The current study: 1) localized IL-6 mRNA levels in preeclamptic versus normal decidual sections; 2) evaluated mechanisms regulating IL-6 synthesis by targeting intracellular signaling pathways with specific inhibitors; 3) identified potential IL-6 targets by immunolocalizing the IL-6 receptor (IL-6R) to specific cell types in placental bed biopsies. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7104449/ doi: 10.1177/19337191080150020102 id: cord-019347-tj3ye1mx author: nan title: ABSTRACT BOOK date: 2010-02-19 words: 107926.0 sentences: 6940.0 pages: flesch: 53.0 cache: ./cache/cord-019347-tj3ye1mx.txt txt: ./txt/cord-019347-tj3ye1mx.txt summary: Method:Case Report:A 15y/o w/f athlete presented with a two month history of recurrent hives and angioedema which she associated with ingestion of Halloween candy .One week before evaluation she had hives with Coconut as well.Her history was othewise unremarkable except for recurrent UTI''S, annual sinusitis, pneumonia in 1998 as well as migraines.She denied sexual activity.Her physical exam was normal.Results:An evaluation for autoimmune disease revealed normal ESR, ANA, DSDNA, mono and hepatitis serology as well as lyme titers however her CH50 was low17u/ml(normal 26-58U/ml)and evaluation of complement revealed c4 14mg/dl(normal 16-47mg//dl)and c2 <1.3mg/dl(normal 1.6-3.5mg/dl)with normal c3, c5-c9.Her father had nor-malc4 but c2 was 1.4mg/dl (normal 1.6-3.5mg/dl)Her sister had c2 of 1.5mg/dl and normal c4 and her mother had normal c2 and c4.Her workup included positive prick skin test to ragweed, ash and grass and she was started on Rhinocort and Clarinex seasonally.She has been followed for one year with resolution of hives and is asymptomatic.Her diagnosis had been confirmed by a pediatric rheumatologist.Conclusion;We present an atypical case of C2 complement deficiency in an currently asymptomatic individual. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7129269/ doi: 10.1016/s1081-1206(10)61294-x id: cord-022501-9wnmdvg5 author: nan title: P1460 – P1884 date: 2015-12-28 words: 128256.0 sentences: 7808.0 pages: flesch: 51.0 cache: ./cache/cord-022501-9wnmdvg5.txt txt: ./txt/cord-022501-9wnmdvg5.txt summary: Methods: Using published data on (1) the prevalence of MRSA and other bacterial pathogens causing cSSSI in the US, (2) the in-vitro susceptibility rates of commonly used regimens in cSSSI in the US in relation to the most pervasive pathogens identified above, and (3) estimated costs of failure of initial, empiric treatment from a recent study of a large US multi-hospital database, we developed a model to predict the expected clinical and economic impact of increasing prevalence of MRSA. Small outbreaks of VEB-1 ESBL producing Acinetobacter baumannii in Belgian nursing homes and hospitals through cross-border transfer of patients from northern France Methods: From 01/04 to 03/05, all Belgian acute hospitals were invited to report cases of nosocomial infections/colonisations due to MDR Ab isolates presenting a resistance profile similar to the French epidemic strain (resistance to all agents except carbapenems and colistin) and to send such isolates to the reference laboratory for phenotypic confirmation and for genotypic characterization (PCR of VEB-1 and class 1 Integron, PFGE typing). abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7157935/ doi: 10.1111/j.1470-9465.2006.12_4_1431.x id: cord-022650-phsr10jp author: nan title: Abstracts TPS date: 2018-08-14 words: 119675.0 sentences: 7010.0 pages: flesch: 55.0 cache: ./cache/cord-022650-phsr10jp.txt txt: ./txt/cord-022650-phsr10jp.txt summary: 0685 | Skin prick test reactivity to aeroallergens in adult allergy clinic in a tertiary hospital: a 12-year retrospective study Results: Five different human sera were screened for specific IgE level against 29 different allergen sources using test methods of three different suppliers. Conclusion: This multicenter prospective study confirmed that stepwise single-dose OFC to egg will help to clarify the severity of egg allergy, and will contribute to improved food allergy manageMethod: The study design was a retrospective cohort study extracting data from the electronic chart of children older than 4 years who visited our out-patient clinic for egg or milk allergy and who underwent an oral food challenge test (OFC) twice within 24 months between November 2013 and December 2017. Results: In the base case analysis, using Italy clinical practice patients with moderate-to severe allergic rhino-conjunctivitis (SS ranging from 6 to 15 points) and a mean age at entry of 21 years, both SCIT and SLIT were associated with increased cost but superior efficacy compared to pharmacotherapy alone. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159469/ doi: 10.1111/all.13539 id: cord-022653-qa1uph35 author: nan title: Poster Discussion Session PDS date: 2017-08-30 words: 58292.0 sentences: 3300.0 pages: flesch: 53.0 cache: ./cache/cord-022653-qa1uph35.txt txt: ./txt/cord-022653-qa1uph35.txt summary: 0206 | G protein coupled receptor kinase 2 (GRK2) regulates endothelial permeability induced by Bradykinin 0208 | Pharmacokinetics (PK) and pharmacodynamics (PD) of c1 esterase inhibitor of chronic urticaria challenges most commonly identified were the following: time of onset of disease; frequency/duration of and provoking factors for wheals; diurnal variation; occurrence in relation to weekends, holidays, and foreign travel; shape, size, and distribution of wheals; associated angioedema; associated subjective symptoms of lesions; family and personal history regarding urticaria, atopy; previous or current allergies, infections, internal diseases, or other possible causes; psychosomatic and psychiatric diseases; surgical implantations and events during surgery; gastric/ intestinal problems; induction by physical agents or exercise; use of drugs; food allergies; relationship to the menstrual cycle; smoking habits; type of work, hobbies; stress; quality of life and emotional impact; previous therapy and response to therapy, and previous diagnostic procedures/results. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159476/ doi: 10.1111/all.13251 id: cord-022888-dnsdg04n author: nan title: Poster Sessions date: 2009-08-19 words: 188640.0 sentences: 9313.0 pages: flesch: 45.0 cache: ./cache/cord-022888-dnsdg04n.txt txt: ./txt/cord-022888-dnsdg04n.txt summary: Methods: Phospho-specific Western blot analyses were performed to verify the functionality of the different IFN-g pathway components, intra-and extracellular flow cytometry experiments were employed to determine the expression of antigen processing components and HLA class I cell surface antigens, quantitative real time-PCR experiments to confirm the absence of JAK2 and presence of pathway relevant molecules as well as, genomic PCR and chromosome typing technique to prove the deletion of JAK2. In order to accomplish these objectives we induced priming or tolerance of ovalbumin (OVA 323-339 peptide)-specific T cells from DO11.10 TCR transgenic mice in vitro or, following adoptive transfer of near physiologically relevant numbers of such cells into recipients, in vivo and correlated functional outcome (via proliferation and cytokine readout assays or antibody production) with E3 ubiquitin-protein ligases expression and the ubiquitination status of the TCR signalling machinery. abstract: No Abtract url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7163517/ doi: 10.1002/eji.200990224 id: cord-022940-atbjwpo5 author: nan title: Poster Sessions date: 2016-09-07 words: 241182.0 sentences: 12746.0 pages: flesch: 47.0 cache: ./cache/cord-022940-atbjwpo5.txt txt: ./txt/cord-022940-atbjwpo5.txt summary: We have studied the effect of inhibition of IRE1 (inositol requiring enzyme 1), which is a central mediator of endoplasmic reticulum stress and controls cell proliferation and tumor growth, on hypoxic regulation of the expression of different proliferation related genes in U87 glioma cells. Transient inhibition of Akt and mTOR protein kinase activation in tumor cells followed by reactivation of signaling pathway did not result in a time-dependent difference on EGFR, HER2 and HER3 expression levels. In our study we aimed to determine cytotoxic effect of RES in K562 human CML cell line and to evaluate the expressions of miRNAs that are associated with genetics of leukemia after treatment with RES; to investigate target genes of miRNAs which show significant expression alterations and molecular mechanisms of RES treatment. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7164006/ doi: 10.1111/febs.13808 id: cord-023095-4dannjjm author: nan title: Research Abstract Program of the 2011 ACVIM Forum Denver, Colorado, June 15–18, 2011 date: 2011-05-03 words: 134226.0 sentences: 6834.0 pages: flesch: 51.0 cache: ./cache/cord-023095-4dannjjm.txt txt: ./txt/cord-023095-4dannjjm.txt summary: The purpose of this study was to determine the short-term effects of ivabradine on heart rate (HR), blood pressure, left ventricular (LV) systolic and diastolic function, left atrial (LA) performance, and clinical tolerance in healthy cats after repeated oral doses. The goal of this study was to investigate the relationship between heart rate and ECG time intervals to body mass in apparently healthy horses and ponies and to calculate normal ranges for different weight groups. This study aimed to investigate the prevalence of hypercoagulability in PLN dogs based on thromboelastography (TEG), and to determine whether hypercoagulability in these patients could be predicted by clinical assessments that identify systemic hypertension (systolic blood pressure 4 160 mmHg), hypoalbuminemia (serum albumin o 2.7 mg/dl), antithrombin activity (o 70%), and degree of proteinuria (urine protein:creatinine ratio [UPC] ! abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7166756/ doi: 10.1111/j.1939-1676.2011.0726.x id: cord-031907-ilhr3iu5 author: nan title: ISEV2020 Abstract Book date: 2020-07-15 words: 200999.0 sentences: 11528.0 pages: flesch: 44.0 cache: ./cache/cord-031907-ilhr3iu5.txt txt: ./txt/cord-031907-ilhr3iu5.txt summary: L.M., and the National Institutes of Health (R35GM119623) to T.R.G. The addition of a size exclusion chromatography step to various urinary extracellular vesicle concentrating methods reveals differences in the small RNA profile Introduction: Urinary extracellular vesicles (EVs) and their RNA cargo are a novel source of biomarkers for various diseases, however non-vesicular RNA (e.g. associated with proteins) is also present within urine. We then evaluated efficiency of heart targeting for eAAV9 or eAAV6 and standard AAV9 or AAV6 encoding for EGFP, mCherry or firefly luciferase in different human cell lines in vitro, in black mouse and in passive immunity nude mouse model in vivo using flow cytometry, confocal microscopy, Langendorff perfusion system and Methods: HLHS patients (n = 3) after Glenn procedure and swine (n = 3) after PAB were given RV injections of allogeneic/xenogeneic MSCs. Donor-specific, HLA-I+, exosomes were isolated from plasma. abstract: nan url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480431/ doi: 10.1080/20013078.2020.1784511 ==== make-pages.sh questions [ERIC WAS HERE] parallel: Warning: Only enough available processes to run 27 jobs in parallel. parallel: Warning: Raising ulimit -u or /etc/security/limits.conf parallel: Warning: or /proc/sys/kernel/pid_max may help. ==== make-pages.sh search /data-disk/reader-compute/reader-cord/bin/make-pages.sh: line 77: /data-disk/reader-compute/reader-cord/tmp/search.htm: No such file or directory Traceback (most recent call last): File "/data-disk/reader-compute/reader-cord/bin/tsv2htm-search.py", line 51, in with open( TEMPLATE, 'r' ) as handle : htm = handle.read() FileNotFoundError: [Errno 2] No such file or directory: '/data-disk/reader-compute/reader-cord/tmp/search.htm' ==== make-pages.sh topic modeling corpus Zipping study carrel