cord-000113-d0eur1hq 2009 The advent of molecular biology and new technological initiatives that combine advances in biology with other disciplines will support the development of techniques capable of high throughput testing with a low turnaround time for rabies diagnosis. The advent of molecular biology and new technological initiatives that combine advances in biology with other disciplines will support the development of techniques capable of high throughput testing with a low turnaround time for rabies diagnosis. Another method for the detection of rabies virus antigen from postmortem samples is a recently developed rapid immunodiagwww.plosntds.org nostic test (RIDT) based on the principles of immunochromatography [13] . Development of RT-LAMP assays for use in diagnosis and surveillance is challenged by the considerable sequence variation observed within the rabies virus genome [44] that can frustrate specific primer design. Currently, high-throughput rabies virus molecular detection methods augment standard diagnostic tests or are in the process of development and refinement for use alone. cord-000283-7s6283y5 2010 We intend to show that these specifically designed Vivo-Morpholinos are effective in countering the viral load in the body, thereby imparting significant protection to the animals that were infected with a lethal dose of JEV. Similarly, E glycoprotein level showed significant increase in JEV-infected and JEV+SC-MO groups when compared to Sham (p,0.01) which were then drastically reduced following 39 and 59 MO treatments (p,0.01) (Figure 3B-D) . CBA performed to check the proinflammatory cytokines levels in the brain homogenates obtained from different treatments showed that levels of MCP-1, IFN-c, TNF-a, and IL-6 were found to be significantly increased in both JEV and JEV+SC-MO groups when compared to Sham infected groups (p,0.01). doi:10.1371/journal.pntd.0000892.g002 Vivo-Morpholino in Japanese Encephalitis www.plosntds.org JEV+SC-MO groups when compared to Sham (p,0.01). Increases were observed in the ROS levels in the brain samples of JEVinfected and JEV+SC-MO groups in comparison to Sham, that were reduced following 39 and 59 MO treatments. cord-000625-cpjlzutk 2012 In order to develop a field applicable technique that offers high detection sensitivity and specificity for the diagnosis of BU, we explored the use of the pocket warmer LAMP (pwLAMP) technique, a DNA amplification method using isothermal conditions (60uC) provided by a disposable pocket warmer [34] . In order to develop a simple and rapid test that can be used to diagnose Buruli ulcer under field conditions, we modified the conventional LAMP assay by using a disposable pocket warmer as a heating device for generating a constant temperature for the test reaction and employed the use of crude sample preparations consisting of boiled and unboiled extracts of the clinical specimen instead of using purified DNA as the diagnostic specimen. Thirty clinical specimens from suspected Buruli ulcer patients were investigated by the modified LAMP (or pocket warmer LAMP) and the conventional LAMP, as well as IS2404 PCR, a reference method for the detection of Mycobacterium ulcerans. cord-000680-vsgd9v1w 2012 We aimed to assess the clinical relevance and discriminatory value of AST or ALT for dengue hemorrhagic fever (DHF) and severe dengue. There was significant overlap in AST and ALT values among patients with dengue with or without warning signs and severe dengue, and between those with DF and DHF. In this study, we aimed to evaluate the clinical relevance of elevated AST and ALT levels and correlate liver aminotransferase levels with dengue severity according to WHO 1997 and 2009 classifications. We performed a subgroup analysis for median maximum AST and ALT values stratified by febrile (days 1-3 of illness), critical (days 4-6), and convalescent (days 7-10) phases as defined by WHO 2009 [1] and compared across dengue severity classification according to WHO 1997 [9] and 2009 [1] . In conclusion, elevated aminotransferase levels were associated with DHF/DSS and severe dengue in our cohort of adult patients with confirmed dengue. cord-001074-qevosik3 2013 C9 was determined to be a potent virus neutralizing antibody and a biosensor antibody binding study demonstrated it recognized residues on intact CHIKV VLPs. Shotgun mutagenesis alanine scanning of 98 percent of the residues in the E1 and E2 glycoproteins of CHIKV envelope showed that the epitope bound by C9 included amino-acid 162 in the acid-sensitive region (ASR) of the CHIKV E2 glycoprotein. The neutralizing epitope bound by C9 mapped to the acid-sensitive region (ASR) that is critical for the rearrangement of CHIKV E2 during fusion and viral entry into host cells. Therefore, C9 is a potent neutralizing antibody that can therapeutically protect wild-type neonate mice from a lethal dose of virus at 8 and 18 hours post CHIKV inoculation. This study describes the isolation and characterization of two human monoclonal antibodies, C9 and E8, from CHIKV infected and recovered individuals. cord-001365-6u80p5sj 2014 cord-001484-va0teako 2014 A rapid, simple, and highly efficient molecular based method for identification of agents of black grain eumycetoma is introduced, aiming to improve diagnostic in endemic areas. Rolling circle amplification (RCA) is a powerful diagnostic method based on detection of specific nucleic-acid sequences and enzymatic amplification of circularized oligonucleotide probes under isothermal conditions [10] . The specificity of the 8 RCA probes was tested using strains of black-grain mycetoma causative species listed in table 1. Identification of mycetoma agent with phenotypic features is too limited, and physiological and biochemical techniques are laborious, time-consuming and nonspecific, whereas the currently available molecular methods based on DNA sequencing are specific but extremely expensive. We describe rolling circle amplification method for identification of black grain eumycetoma using species-specific padlock probes. In the present study we developed a simple, fast and highly specific molecular method for the identification of agents of black grain mycetoma. cord-001642-bom9fk1y 2015 We showed that AE-fgl2(-/-) mice exhibited (as compared to AE-WT-animals) (a) a significantly lower parasite load with reduced proliferation activity, (b) an increased T cell proliferative response to ConA, (c) reduced Treg numbers and function, and (d) a persistent capacity of Th1 polarization and DC maturation. multilocularis-infected fgl2 -/mice and control WT littermates were analyzed after 1 and 4 months p.i. with respect to parasite weight and the expression of em14-3-3 as a marker for cellular proliferation activity [24] . Quantitative RT-PCR showed that fgl2 mRNA-levels were significantly increased in CD4 + CD25 + Tregs derived from AE-WT mice, when compared to non-infected controls, while no significant changes were evident with respect to CD4 + Teffs, CD8 + T cells. To further explore the role of VF on Tregs and FGL2 secretion, respectively, spleen cells from AE-WT mice and non-infected WT controls were each cultured in the presence of VF (10 μg/mL). cord-001690-cn21fgug 2015 In the present study, a new vaccination strategy approach based on three recombinant bovine herpesvirus 4 (BoHV-4) vectors, each expressing different MPXV glycoproteins, A29L, M1R and B6R were investigated in terms of protection from a lethal MPXV challenge in STAT1 knockout mice. In vivo efficacy testing of BoHV-4-A-CMV-A29LgD 106 ΔTK, BoHV-4-A-EF1α-M1RgD 106 ΔTK and BoHV-4-A-EF1α-B6RgD 106 ΔTK To test the efficacy of the vectors in vivo, we sought to determine if they could protect mice against a lethal challenge with MPXV. Since the purpose of this study was to determine the capability of BoHV-4-based viral vectors to protect STAT1 (-/-) mice against a lethal MPXV infection, the first concern was the generation of optimized expression cassettes to be integrated into the BAC-BoHV-4-A genome that were able to efficiently express A29L, M1R and B6R antigens. In summary, our findings have demonstrated that BoHV-4 based vectors can be used as vaccines to protect against a lethal MPXV challenge in mice. cord-001812-ov1qssnu 2015 cord-002179-v8lpw4r7 2016 title: Identification of RNA Binding Proteins Associated with Dengue Virus RNA in Infected Cells Reveals Temporally Distinct Host Factor Requirements Previously, we have described a high-throughput mass spectrometry method termed TUX-MS (thiouracil cross-linking mass spectrometry) to identify host factors that interact with viral RNA during a live infection in cell culture [15] . Development of a quantitative thiouridine cross-linking mass spectrometry (qTUX-MS) method for identification of proteins associated with the DENV RNA TUX-MS can be used to identify host factors by incorporating 4-thiouridine (4sU), a zero-distance cross-linker, into the viral RNA (vRNA) to enable cross-linking of proteins bound to vRNA during a live infection in cell culture [15] . Since some of hnRNPs are known to modulate cellular gene expression in response to dengue infection [60, 61] , we can not rule out that some of the observed effects on virus titers derive from their roles in regulating host mRNAs. Among the numerous qTUX-MS identified factors of interest, our study is the first to demonstrate the involvement of HMCES (or C3orf37) in viral infection. cord-002248-92pzqj35 2016 Importantly, RVFV MP-12-NSs mutant viruses with an impaired general transcription inhibition function showed a reduced cytotoxicity in cell culture and attenuated virulence in young mice, compared with its parental virus MP-12, highlighting the contribution of NSs-mediated general transcription inhibition towards RVFV virulence. Each carried mutations in NSs that specifically targeted its general transcription inhibition function without affecting its ability to degrade PKR and inhibit IFN-β promoter induction, through its interaction with Sin3-associated protein 30, a part of the repressor complex at the IFN-β promoter. Importantly, RVFV MP-12-NSs mutant viruses with an impaired general transcription inhibition function showed a reduced cytotoxicity in cell culture and attenuated virulence in young mice, Both NSs mutants retained some NSs functions, including degradation of PKR and SAP30 binding, and yet these NSs mutants and wt NSs showed different levels of general transcriptional suppression activities in virus-infected cells. cord-002394-n85ptr5p 2017 Therefore, in this study we investigated the possible occurrence of molecular mimicry between CHIKV E1 and host components using a three pronged strategy: (i) identification of homologous regions between CHIKV proteins and host tissue components using bioinformatics tools, (ii) establishing cross reactivity between serum samples obtained from CHIKV infected patients and peptides exhibiting molecular mimicry and (iii) validating the ability of the cross reactive peptides in inducing joint and muscle pathology in a mouse model. We investigated molecular mimicry in this study by using a combined approach of identifying homologous regions between CHIKV glycoprotein E1 protein and host tissue components using bioinformatics tools, the ability of these designed peptides to cross react with serum samples from CHIKV infected patients and inducing immune mediated joint and muscle pathology in a mouse model. cord-002581-r7mskri0 2017 title: A human inferred germline antibody binds to an immunodominant epitope and neutralizes Zika virus The isolation of neutralizing monoclonal antibodies (nmAbs) against the Zika virus (ZIKV) might lead to novel preventative strategies for infections in at-risk individuals, primarily pregnant women. Here we describe the isolation of 18 plasmablast-derived human mAbs, sorted 12 days post onset of symptoms from a ZIKV-patient in São Paulo, Brazil. Interestingly, one of these mAbs (P1F12) exhibited no nucleotide mutations when compared to its corresponding germline sequences, but still recognized a ZIKV immunodominant epitope and neutralized the virus. Virus capture assay and recombinant E protein ELISA P1F12 binding was determined by both virus capture assay (VCA) and recombinant (r)E ELISAs. The VCA plates were coated overnight with the mouse-anti-Flavivirus monoclonal antibody 4G2 (clone D1-4G2-4-15, EMD Millipore) followed by incubation with viral stocks (ZIKV or DENV). Molecular determinants of human neutralizing antibodies isolated from a patient infected with Zika virus cord-003006-lk2ny1wd 2018 These newly discovered roles are revealing new mechanisms of virus replication and pathogenicity, whilst enhancing our understanding of the broad functions of each ebolavirus viral protein (VP). Lastly, VP35 is a suppressor of RNA silencing, functionally equivalent to the human immunodeficiency virus (HIV-1) Trans-activator of transcription (Tat) protein and important for viral evasion of the innate immune response [34] . The authors propose that this mutation is likely a result of EBOV adaptation to the human host, as several viral variants have been seen to increase human cell infectivity while decreasing virus entry in nonhuman primates [83] [84] [85] . In addition, many ebolavirus proteins are seen to interact with immune cells, causing cell activation and/or cell death and facilitating both viral replication and spread (e.g., by recruiting monocytes to infected cells or by increasing vascular leakage) as well as enabling immune evasion (e.g., antibody neutralisation by sGP and by cleaved GP), roles that were previously solely attributed to VP24 and VP35. cord-003243-u744apzw 2018 cord-003389-0yh5k6jk 2018 cord-003482-f1uvohf0 2019 Quantitative probe-based reverse transcription PCR (qRT-PCR) was performed on seruminoculated Vero cell supernatants, serum, brain, lung, liver, spleen, kidney, urinary bladder, prostate and testes from bats from both studies. Brain and testicular tissues stained with both goat polyclonal goat anti-Iba1 (green) and monoclonal 4G-2 flavivirus E specific antibodies (red) showed co-localization (yellow) of ZIKV antigen in cytoplasm of activated microglial cells with their characteristic morphology in the cerebral cortex of infected bats 10 dpi in the time course study and 28 day dpi in the pilot study (Fig 9) . Two bat infection experiments were conducted in this investigation; 1) a pilot study to determine susceptibility of Jamaican fruit bats to ZIKV infection, and 2) a time course study to better understand pathophysiology and chronology of events pertaining to the dynamics of viremia, viral tropism, replication and shedding of the virus in a New World bat species. cord-004247-lagv3tp7 2020 This mAb was protective in an in vitro, antigen-dependent, cellular cytotoxicity assay with rat macrophages or eosinophils and also in vivo during the early phase of infection Second, beyond the cell-surface proteins, schistosomes also express a large number of glycans as part of their glycoprotein and glycolipid repertoire, and an antibody response against those glycans is mounted by the infected host [80] . In addition to antibodies that directly target and inhibit the fungal pathogen, mAbs can be directed to checkpoints that control the host immune response. In addition to highlighting the potential of mAbs as therapeutics, these studies have demonstrated the diversity of inhibitory actions that mAbs can perform on cryptococcal cells, which can include opsonization and increased phagocytosis, inhibition of fungal growth, capsular polysaccharide release and biofilm formation, antibody-mediated target cleavage, and augmentation of the host response [104] [105] [106] [107] . cord-007383-5yb3dxse 2020 title: Vaccination with single plasmid DNA encoding IL-12 and antigens of severe fever with thrombocytopenia syndrome virus elicits complete protection in IFNAR knockout mice Immunization of NS antigen with Freund''s adjuvant in C57BL/6 mice, which are naturally resistant to SFTSV but partially mimic human infections [12] , failed to enhance viral clearance, although it induced high titer of anti-NS antibodies and significantly elevated IFN-γ levels in sera upon viral challenge [9] . Vaccination of pSFTSV-IL12 provided complete protection of IFNAR KO mice upon lethal SFTSV challenge, whereas immunization with pSFTSV elicits only partial protection, indicating that antigen-specific cellular immune responses enhanced by co-expression of IL-12 could play a significant role in protection against lethal SFTSV infection. Since we observed significant elevation of T cell responses specific to the viral antigens in IFNAR KO mice immunized with pSFTSV-IL12 DNA vaccine, we tested whether it could provide protective immunity against lethal SFTSV infection. cord-012911-d6ct94d9 2020 In August 2014, the Government of Sierra Leone repurposed an existing national, toll-free telephone line (1-1-7 system) for communities to report all deaths and suspected Ebola patients as part of the epidemic response [13] . To inform strategies for improving routine surveillance of deaths, in April 2017, we then conducted a cross-sectional, telephone-based survey with individuals aged 18 years and above who reported a death to the 1-1-7 system after the end of enhanced Ebola surveillance in Sierra Leone. The monthly aggregated data of deaths reported to the 1-1-7 system showed a sharp decline after the 2014-2015 Ebola epidemic ended in Sierra Leone compared to the post-outbreak enhanced mortality surveillance period. In the telephone survey we identified motivations related to death reporting that have practical implications for improving routine mortality surveillance in a post-Ebola-outbreak setting. cord-256303-bpa571ys 2020 cord-260412-yjr83ef6 2020 cord-260693-8mfuwx8l 2020 However, a similar approach should also be adopted for the control of arboviral diseases of global importance, including dengue, Zika, chikungunya, and yellow fever, as recommended by the Pan-American Health Organization (PAHO) in their interim guidance on control of Aedes aegypti mosquitos during the COVID-19 pandemic [2] . The combined impact of both COVID-19 and epidemics of dengue or other vector-borne diseases (VBDs) could have potentially devastating consequences [6] . • Continue the implementation of the WHO''s global vector control response 2017-2030 (GVCR) strategy and regional policies for vector control [7, 8] , with respect to inter-and intrasectoral collaboration, engagement and mobilisation of communities, and scaling up of vector control if required, according to the implementation plan of vector control activities, while adapting activities as necessary to prevent further spread of COVID-19, in particular vector surveillance, which may need to be scaled down [9, 10] . It is vital that the COVID-19 response does not increase VBD threats in these communities by derailing global vector control efforts. cord-263044-o8aosx2q 2015 cord-268329-apl6n6jl 2020 The coronavirus disease 2019 (COVID-19)-caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a betacoronavirus (betaCoV)-emerged for the first time as an outbreak of pneumonia in Wuhan, China, and it is now spreading to several countries around the world [1] . Some studies of SARS-CoV-2 infection have reported the presence of a cytokine storm syndrome and a subgroup of patients who progressed to severe forms of the disease, expressing a pro-inflammatory profile in plasma with IL-2, IL-7, TNF-α, and others as significant complications, such as occurs in T1R [10, 11] . In both reactions, we warn of the possible effect that COVID-19 infection may have on the number of cases of these immunological events because the presence of infection is an important risk factor for triggering leprosy reactions [8] . Another disturbing factor, which may contribute to the susceptibility of those affected by leprosy reactions, are the treatments implemented during these events that interfere with the inflammatory response of these patients. cord-269004-hj03n13h 2010 cord-270481-rrpqz0uy 2020 cord-272250-asuxx1ln 2011 cord-276271-3nz3169p 2009 cruzi OligoC-TesT: A Simplified and Standardized Polymerase Chain Reaction Format for Diagnosis of Chagas Disease The specificity and sensitivity of the assay were evaluated on blood samples from 60 Chagas non-endemic and 48 endemic control persons and on biological samples from 33 patients, 7 reservoir animals, and 14 vectors collected in Chile. We assessed the diagnostic accuracy of the assay on a broad spectrum of biological samples from Chagas non-endemic and endemic control persons, infected children, adults, reservoir animals and vectors. DNA from 25 different Trypanosoma rangeli isolates (Table 1) and from Leishmania donovani, Trypanosoma brucei gambiense, Mycobacterium tuberculosis, Schistosoma mansoni and Plasmodium falciparum was obtained from other research groups. cruzi OligoC-TesT, a simple and standardized dipstick format for detection of PCR amplified T. cruzi OligoC-TesT, a simple and standardized dipstick format for detection of PCR amplified T. cord-276850-tnlyk0wz 2015 cord-280030-neqycg6v 2014 cord-281456-3dlsbr7c 2014 cord-285772-4xt4anq5 2020 cord-286255-ded5t1ai 2017 Blood and oro-nasopharyngeal specimens were tested by RT-PCR and immunodiagnostic methods for infection with dengue viruses (DENV) 1–4, chikungunya virus (CHIKV), influenza A and B viruses (FLU A/B), 12 other respiratory viruses (ORV), enterovirus, Leptospira spp., and Burkholderia pseudomallei. Clinical predictors of laboratory-positive dengue compared to all other AFI etiologies were determined by age and day post-illness onset (DPO) at presentation. By enrolling febrile patients at clinical presentation, we identified unbiased predictors of laboratory-positive dengue as compared to other common causes of AFI. Among 6,349 participants who presented early (<3 DPO) in the clinical course, leukopenia, thrombocytopenia, headache, eye pain, nausea, and dizziness were significant positive predictors of laboratory-positive dengue as compared to all other AFI cases across all age groups ( Table 5 ). Of note, 6% of participants !65 years old had dengue as a cause of AFI, a finding comparable to a Puerto Rico study in which 5% of 17,666 laboratory-positive dengue cases detected by surveillance were !65 years old [36] . cord-287582-ya81rc2n 2016 We identified important epidemiological and environmental issues including the increase of local transmission despite control efforts, population growth, difficulty locating larval sites, and increased human mobility from neighboring endemic countries. A robust assessment of the economic burden helps to i) identify information gaps, research needs, and refinements to the national statistical reporting systems; ii) argue that policies on dengue control and prevention should be given a high priority on the public health policy agenda; and iii) provide a baseline measure to determine the cost effectiveness of dengue policies and programs [104] . The countries and states presented in this review have in common i) a competent mosquito vector, ii) introduction of DENV by travelers, iii) increased local transmission of dengue coinciding with population growth and increased mobility, iv) recent budget cuts impacting public health services, and v) a largely nonimmune population. cord-288202-r3r2bc7v 2013 cord-292157-hrm69640 2020 Typhimurium infection and antibiotic treatment failure, we assessed the potential of two consecutive doses of vitamin A in alleviating infection in male and female mice on a VAD or control diet. We found that subtherapeutic antibiotic treatment synergized with vitamin A treatment in infected VAD male mice, significantly decreasing systemic bacterial levels, mitigating weight loss and improving survival. Typhimurium systemic bacterial levels (CFU) were assessed for male (n = 5) and female (n = 5) mice on a standard diet 5 days post-infection for the following treatment groups: 0 mg/ml, 0.01 mg/ml, 0.05 mg/ml, and 0.10 mg/ml enrofloxacin delivered in the drinking water. Typhimurium D23580 at day 4 post-infection were assessed for male and female mice on either control or VAD diets with the following treatment groups: mocktreated, vitamin A only, enrofloxacin (0.05 mg/ml) only, and vitamin A and enrofloxacin cotreatment (Fig 5A) . cord-294798-ji3p0l4j 2018 We have previously reported isolation of ZIKV [8] , DENV [9] , Human coronavirus NL63 [10] , and Enterovirus D68 [11] from children in this school cohort; however, the cases/outbreaks in these prior publications did not include CHIKV, or cases within the May-August, 2014, time frame of the current study. Viral genomic RNA that was extracted from CHIKV, DENV1-4, and ZIKV strains that were obtained from the Biodefense and Emerging Infections Research Resource Repository (BEI Resources, Manassas, VA) were used as positive control materials for rtRT-PCR. The six ZIKV sequences obtained in June 2014 from the CHIKV co-infected patients were highly similar (99.9%) to each other and also cluster within a well-supported monophyletic clade, which, interestingly, includes a divergent strain isolated in 2016 from Guadaloupe (Figs 3B and S3B) . cord-296226-ugeupo3u 2020 Aedes-borne diseases, in particular, including dengue, chikungunya, yellow fever, and Zika, are increasing at an alarming rate due to urbanisation, population movement, weak vector control programmes, and climate change. The environmental management put in place to implement this high standard of public cleanliness has greatly benefited Singapore''s efforts to tackle VBDs. Underscoring the view that Aedes-borne diseases are environmental diseases, dengue control in Singapore is led by the National Environment Agency (NEA), a statutory board of the Ministry of the Environment and Water Resources (MEWR). In view of the importance of infrastructure maintenance and design, environmental sanitation, people''s behaviours, and use of technologies on dengue prevention, the NEA collaborates closely with other government ministries (e.g., Health, National Development, Education, Finance), town councils (responsible for management and maintenance of the common property of public housing estates, including vector control), community associations, research and academic institutions, and the private sector (Fig 2) . cord-303647-c4umbcvn 2014 cord-305890-mdwjrfzp 2010 cord-306952-cpltrsa7 2020 cord-308343-crjjhpl1 2018 cord-309587-xc4jaw31 2010 cord-310870-w8wu8vno 2017 cord-312223-qgwzgazd 2013 cord-322943-lvdl7puw 2010 cord-327799-ngzvdd8c 2020 cord-332473-ec8lu2a3 2017 cord-340569-f1odmjcs 2020 cord-340939-ikomc19t 2019 cord-345315-y3bdjnhg 2020 We performed a comparative study to determine the feasibility of the early detection of the COVID-19 outbreak in China based on influenza surveillance data and the internet-based Baidu search index to evaluate the timelines of the alert signals compared with the traditional case reporting and response systems. The findings from this study suggest that monitoring abnormal surges of ILI and identifying peaks of online searches of key terms can provide early signals of novel disease outbreaks. In this study, we performed a comparative study to discuss the early warning capability, timelines, and validity of alert signals for the first wave of the COVID-19 outbreak in China based on the surveillance data of influenza-like illness (ILI) and the Baidu Search Index (BSI) compared with the traditional case reporting system. cord-345494-8lcdx719 2015 title: Development of Recombinase Polymerase Amplification Assays for Detection of Orientia tsutsugamushi or Rickettsia typhi Recombinase polymerase amplification (RPA) assays using a lateral flow test (RPA-nfo) and real-time fluorescent detection (RPA-exo) were developed targeting the 47-kDa gene of O. Almost all antigen/pathogen detection assays for rickettsial diseases are based on polymerase chain reaction (PCR [7, 8] ), quantitative real-time PCR (qPCR) or nested PCR targeting different genes, including 56 kDa [9] , 47kDa [10] , groEL [11] of O. The DNA was used as template for the 17-RPA assay to evaluate its performance and qPCR was used to quantitate the copy number of the 17 kDa gene. honei and R japonica were also evaluated to show that they were not detectable by 17-RPA-nfo unless 10 4 or more copies of DNA were present, same results were obtained using 17-RPA-exo. cord-350533-fp1ctpax 2020 cord-352178-irjhmxsg 2013 Fifteen distinct chimeric viruses were constructed to evaluate both structural and non-structural regions of the genome and infection patterns were determined through artificial infectious feeds in An. gambiae with each of these chimeras. When ONNV non-structural protein 3 (nsP3) replaced nsP3 from CHIKV virus in one of the chimeric viruses, infection rates in An. gambiae went from 0% to 63.5%. Our study analyzed both structural and non-structural regions of the ONNV genome using chimeric viruses and artificially infected Anopheles gambiae mosquitoes. When ONNV non-structural protein 3 (nsP3) replaced nsP3 from chikungunya virus in one of the chimeric viruses, infection rates in An. gambiae went from 0% to 63.5%. Six additional non-structural chimeric viruses were also constructed using a novel type II restriction enzyme cloning strategy to examine the broader genome with respect to ONNV''s unique vector specificity for An. gambiae mosquitoes (Figure 2) . cord-352771-s0hfsxzb 2016 Based on our previous homology model structure for Gc from Andes hantavirus (ANDV), here we predicted and generated recombinant DIII and stem peptides to test whether these fragments inhibit hantavirus membrane fusion and cell entry. The Gc fragments interfered in ANDV cell entry by preventing membrane hemifusion and pore formation, retaining Gc in a non-resistant homotrimer stage, as described for DIII and stem peptide inhibitors of class II fusion proteins. Once the predicted DIII and stem fragments were synthesized, purified, and characterized, we measured their inhibitory activity against ANDV during virus cell entry via the native, endosomal infection route. In addition to the DIII of ANDV, peptides derived from the stem region of ANDV also cross-inhibited the PUUV fusion activity, which further corroborates the presence of conserved residues among hantavirus Gc proteins that are involved in the likely binding of this peptide. cord-355469-ojop6i4k 2017