cord-000434-ff2zadol	2011	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.
cord-001446-mpuovmeb	2014	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
cord-001521-l36f1gp7	2011	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.
cord-003208-lwirkob3	2018	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.
cord-003315-r1wkx0ml	2018	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.
cord-003471-tr3ageky	2019	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.
cord-003623-n01rgqyv	2019	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).
cord-003656-7mzsaz7a	2019	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.
cord-003859-k8wfyj9b	2019	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.
cord-004101-0r2g5p1i	2020	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
cord-006828-i88on326	2013	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
cord-006860-a3b8hyyr	1996	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.
cord-007480-grndfx7b	2002	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).
cord-007495-gpz4gkv3	2002	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.
cord-007497-nn5l5rai	2002	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.
cord-007644-7bsixsgd	2000	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.
cord-009784-kxa68zbc	2020	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.
cord-009877-3cyz6o9c	2005	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.
cord-009922-t1hoox6e	2005	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).
cord-010247-cug21fnf	1992	
cord-010578-uib9h1lb	1995	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
cord-011212-ovjdzyxv	2019	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.
cord-011840-neowfhwg	2020	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
cord-012891-heqsfzkm	2020	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.
cord-013348-lsksys56	2020	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.
cord-014462-11ggaqf1	2011	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.
cord-014965-efmozngq	2001	
cord-015021-pol2qm74	1994	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.
cord-015147-h0o0yqv8	2014	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.
cord-015394-uj7fe5y6	2008	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.
cord-015683-a9a82of4	2016	
cord-019347-tj3ye1mx	2010	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.
cord-022310-yc6xtw0s	2011	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.
cord-022353-q2k2krnm	2007	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.
cord-022501-9wnmdvg5	2015	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).
cord-022650-phsr10jp	2018	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.
cord-022653-qa1uph35	2017	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.
cord-022888-dnsdg04n	2009	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.
cord-022940-atbjwpo5	2016	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.
cord-023095-4dannjjm	2011	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] !
cord-024171-x7y1xpsf	2010	
cord-025922-84pheilu	2020	
cord-026559-xx52u01h	2020	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.
cord-031190-4qpnlgb5	2020	
cord-031907-ilhr3iu5	2020	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.
cord-032689-2xtiiejf	2020	
cord-251943-jzaeaxam	2005	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.
cord-252867-lku0cm62	1987	
cord-254318-w8wrn9lx	2020	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.
cord-254384-mwzz1db5	2011	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.
cord-255390-dvp0luxe	2000	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.
cord-257190-iesysf3l	2005	
cord-257715-pbcr81qm	2009	
cord-261372-xjbs09gi	2010	
cord-262268-gm99cadh	2003	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.
cord-262640-4vr4cm1s	2020	
cord-262762-mgegswvn	1982	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.
cord-264936-3posyr5n	2014	
cord-264968-ctx39vhi	2004	
cord-265312-yfjme53q	2019	
cord-267736-rya9w6sh	2012	
cord-273361-i5v5rz4x	2013	
cord-275793-k0uvqcmp	2010	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
cord-276989-441aclcc	2020	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.
cord-277186-sj8ngpk8	2005	
cord-277265-p8pns7r9	2019	
cord-277735-a9gkath5	2004	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) .
cord-278324-eqqvwwh6	2018	
cord-279406-wwdqh9qs	2020	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] .
cord-280939-d478p8u6	2020	
cord-281081-rifr5uub	2020	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.
cord-281101-gv1sgbk1	2006	
cord-281393-96j70n2z	2020	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.
cord-281760-34wuttqw	2019	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
cord-284045-scd3f8vk	2020	
cord-284841-flhfagp3	2020	
cord-285700-9q6vwoct	2020	
cord-287590-jjft3den	2005	
cord-288202-r3r2bc7v	2013	
cord-289413-mbrw85og	2019	
cord-290705-7xkt6u73	2020	
cord-291104-6chpmgry	2016	
cord-293393-kbndie8e	2014	
cord-293770-n4ooziv4	2008	
cord-294478-3ickafd3	2008	
cord-297684-9q3oopaz	2020	
cord-298766-xqkre25z	2010	
cord-299148-uge5uodk	2020	
cord-299189-59d4aojh	2013	
cord-300701-vkzya7uq	1987	
cord-300908-i80tuhqk	2015	
cord-301175-6alsigxk	2015	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.
cord-301313-9595vm0k	2020	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).
cord-301355-9lswjro2	2015	
cord-301655-6nxhvvm4	2019	
cord-301974-4wn40ivq	2004	
cord-305516-s1lvhknm	2008	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.
cord-306390-pzzev8hd	2020	
cord-307110-eiobmxp2	2019	
cord-309415-77b5erfj	1987	
cord-310536-u30cufg7	2018	
cord-311349-145kwny3	2014	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.
cord-311811-nrodyagi	2019	
cord-312456-6lxc2rj2	2016	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.
cord-313193-q5zeoqlb	2020	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
cord-313517-5ipj2z86	2019	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] .
cord-316723-srenbxa7	2004	
cord-317026-9zgc6xrb	2019	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 ).
cord-317057-c2bwky6e	2020	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.
cord-317123-0tdfvlqd	2020	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).
cord-317223-juw4xt8q	1986	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.
cord-318266-i8x80knq	1994	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.
cord-318444-sgm24q1i	2020	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.
cord-320559-up1q3k6q	2018	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) .
cord-321691-46la29tm	2004	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).
cord-322529-3xn5v54s	2004	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) .
cord-326232-668f53qc	2018	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.
cord-327890-ocisq7e4	2020	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.
cord-331277-fjsuo3yy	2020	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.
cord-333026-9f6ecg30	2017	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.
cord-334165-7gfk554m	2020	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.
cord-334277-g3go3u02	2020	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.
cord-334896-3g75spkc	2016	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).
cord-334968-gonx5taq	2013	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.
cord-335121-ro3x3qa3	1988	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.
cord-335591-r0x8yaqj	2007	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
cord-336899-zngp67tb	2020	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.
cord-340960-abanr641	2020	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).
cord-342024-kaku49xd	2020	• 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.
cord-343185-lbmbp9ca	2020	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.
cord-344309-6c2wttxg	2018	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
cord-344581-h7ikjgic	2020	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.
cord-344871-486sk4wc	2016	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.
cord-346320-ysgz6adr	2016	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) .
cord-346574-u28y1ttw	2012	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.
cord-347374-mryazbnq	2020	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.
cord-348161-757c51xw	2007	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.
cord-348660-qnbgywgy	2018	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.
cord-349744-8cg5yj20	2020	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.
cord-351498-bmq6zcb0	2018	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.
cord-351952-lhhjax3s	2020	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) .
cord-353190-7qcoxl81	2012	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.
cord-353614-z4fpy607	2007	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.