Journal of virological methods最新文献

{"title":"Evaluation of the Seegene Allplex™ RV master assay for one-step simultaneous detection of eight respiratory viruses in nasopharyngeal specimens","authors":"Anele Mdunyelwa ,&nbsp;Colette Seema ,&nbsp;Anna Mabaso ,&nbsp;Khamusi Mlambo ,&nbsp;Mandisa Mtsweni ,&nbsp;Mathapelo Maphanga ,&nbsp;Elizabeth Rammutla ,&nbsp;Hugo A. Tempelman ,&nbsp;Chijioke N. Umunnakwe","doi":"10.1016/j.jviromet.2024.115042","DOIUrl":"10.1016/j.jviromet.2024.115042","url":null,"abstract":"<div><h3>Background</h3><div>The Seegene Allplex™ RV Master (RVM) assay is a one-step multiplex real-time reverse transcription polymerase chain reaction (RT-PCR) system for detecting eight viral respiratory pathogens from nasopharyngeal swab, aspirate, and bronchoalveolar lavage specimens. The eight RVM targets are: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Influenza A (Flu A), Influenza B (Flu B), Human respiratory syncytial virus (RSV), adenovirus (AdV), rhinovirus (HRV), parainfluenza virus (PIV), and metapneumovirus (MPV). The assay is based on Seegene’s multiple detection temperature (MuDT) technology and provides cycle threshold (Ct) values for each of its viral targets upon PCR completion.</div></div><div><h3>Objective</h3><div>We aimed to evaluate the diagnostic performance of the RVM assay by calculating sensitivity, specificity, accuracy, Positive Predictive Value (PPV), Negative Predictive Value (NPV), Positive Percent Agreement (PPA), Negative Percent Agreement (NPA), and Overall Percent Agreement (OPA) compared to definite diagnosis and analogous reference assays.</div></div><div><h3>Study design</h3><div>Diagnostic sensitivity, specificity, accuracy, PPV, and NPV were calculated by comparing the results of the RVM assay to that of definite diagnosis assays; while PPA, NPA, and OPA were calculated by comparing results of the RVM assay to that of analogous reference products. Definite diagnosis and reference methods comprised whole genome sequencing and PCR genotyping, the Allplex™ SARS-CoV-2/FluA/FluB/RSV and Respiratory Panels 1, 2, and 3 assays (Seegene), and the Xpert® Xpress SARS-CoV-2/FluA/FluB/RSV Plus assay (Cepheid). Reproducibility of the RVM assay using fully-automated and semi-automated nucleic acid (NA) extraction workflows and as performed by independent operators was also assessed. In total, 249 positive respiratory specimens and at least 50 negative specimens for each target tested were used for this evaluation study.</div></div><div><h3>Results</h3><div>Sensitivity, specificity, accuracy, PPV, NPV, PPA, NPA, and OPA ranged from 95.7 % to 100 % for detecting all eight targets tested on the RVM assay. Reproducibility PPA, NPA, and OPA between automated and semi-automated NA extraction workflows were all &gt;97.9 %, while the reproducibility PPA, NPA and OPA between independent operators were all 100 %.</div></div><div><h3>Conclusion</h3><div>These results demonstrate a high level of sensitivity, specificity, accuracy and diagnostic predictive value of the RVM assay and high agreement with comparable reference assays in identifying all eight of its targets. Taken together, our study underscores the diagnostic utility of the RVM assay in detecting eight viral respiratory pathogens.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115042"},"PeriodicalIF":2.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Validation of an HPLC-CAD method for measuring the lipid content of novel LNP-encapsulated COVID-19 mRNA vaccines","authors":"Huan Yang ,&nbsp;Chengrui Fei ,&nbsp;Sijie Wang ,&nbsp;Xue Shen ,&nbsp;Li Yang ,&nbsp;Hefeng Yang ,&nbsp;Guiding Li","doi":"10.1016/j.jviromet.2024.115040","DOIUrl":"10.1016/j.jviromet.2024.115040","url":null,"abstract":"<div><div>Lipid nanoparticles (LNPs) are frequently employed as mRNA vaccine delivery vehicles. LNPs are made up of four types of lipids: cationic lipid, PEG-lipid conjugate, zwitterionic helper phospholipid, and cholesterol. LNP distribution efficiency is significantly impacted by lipid composition, which also controls LNP stability and bilayer fluidity. The various lipids used in the formulation system have distinct properties and contents. To aid in the development of new drugs and vaccines, we developed and validated an HPLC-CAD method for identifying and determining the amounts of four lipids in Yuxi Watson Biotechnology Co., Ltd.'s LNP-encapsulated COVID-19 mRNA vaccines (OmicronXBB.1.5).</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115040"},"PeriodicalIF":2.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surveillance of human adenoviruses in water environments: Assessing the suitability of a locally developed quenching of unincorporated amplification signal reporters-loop-mediated isothermal amplification assay","authors":"Joy Ann P. Santos ,&nbsp;Elchin Juanico ,&nbsp;Joseth Jermaine Abello ,&nbsp;Jonah L. Bondoc ,&nbsp;Windell L. Rivera","doi":"10.1016/j.jviromet.2024.115041","DOIUrl":"10.1016/j.jviromet.2024.115041","url":null,"abstract":"<div><div>The human adenovirus (HAdV) has shown greater environmental persistence, more water treatment resistance than bacteria, and cause infection even at low concentrations. HAdV causes gastrointestinal illnesses and is abundant in various environmental samples such as groundwater, surface water, recreational water, and drinking water. The detection of these pathogens calls for a more practical and affordable approach. A recent technology based on the quenching of unincorporated amplification signal reporters (QUASR) was adapted for the detection of human adenoviruses. This technology allows for non-inhibitory, single-step DNA detection in a closed tube. The QUASR-(loop-mediated isothermal amplification (LAMP) assay was previously optimized and was tested for its applicability to detect enteric HAdV in two areas where people can unintentionally be exposed to contaminated water. A total of 203 water samples were collected and tested using both real-time PCR and QUASR-LAMP assays. Results showed a higher positivity rate of 78.82 % (160/203) for QUASR-LAMP compared to qPCR with only 58.62 % (119/203). The sensitivity and specificity rates for QUASR-LAMP were calculated at 86.55 % and 32.14 %, respectively, when compared to qPCR. The QUASR-LAMP assay's ability to detect target analytes even at low concentrations can be attributed to its increased diagnostic sensitivity but lower specificity since there were samples that were positive in PCR but negative in the QUASR-LAMP assay. However, this characteristic does not diminish its utility as a valuable tool for the detection of HAdV. In fact, this attribute enhances its advantages in situations with constrained space and instrumentation requirements, making it suitable for rapid surveillance of important viruses. Finally, the capacity of the QUASR-LAMP assay to differentiate between positive and negative samples at a defined endpoint is highly beneficial for laboratory technicians who possess limited molecular biology expertise or experience. The QUASR-LAMP platform has demonstrated its usefulness as a diagnostic tool for surveillance of enteric adenoviruses in water sources.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115041"},"PeriodicalIF":2.2,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of norovirus VLPs of the nine representative genotypes widely distributed in Japan using the silkworm-baculovirus expression vector system","authors":"Yuto Tsurumi ,&nbsp;Keisuke Morimoto ,&nbsp;Akitsu Masuda ,&nbsp;Jae Man Lee ,&nbsp;Hiroaki Mon ,&nbsp;Takahiro Kusakabe","doi":"10.1016/j.jviromet.2024.115038","DOIUrl":"10.1016/j.jviromet.2024.115038","url":null,"abstract":"<div><div>Norovirus (NoV) is one of the major causes of acute viral gastroenteritis in humans. Genetic variation is abundant, and prevalent genotypes vary from year to year and region to region. Since NoVs are difficult to amplify in cultured cells, genome RNA-free virus-like particles (VLPs) that mimic the capsid structure of the virus are promising vaccine candidates for the prevention of NoVs infection, and the development of multivalent VLP vaccines is required to prevent NoV infection in a wide range of genotypes. In this study, we attempted to produce NoV VLPs of the top nine genotypes that have a history of epidemics in Japan using the silkworm-baculovirus expression vector system (silkworm-BEVS), which has a proven track record in the mass production of recombinant proteins. In silkworm pupae infected with recombinant baculoviruses constructed to express VP1s, the major protein that forms VLP, the NoV VP1 protein was expressed in large amounts. Most genotypes of VP1 accumulated in the cytoplasm as soluble proteins, but solubility was reduced for that of two genotypes. VP1s of five genotypes could be purified in large quantities (&gt;0.9 mg per pupa) by a two-step purification process, and gel filtration chromatography analysis confirmed the formation of VLPs. This study demonstrates the utility of silkworm-BEVS in producing NoV VLPs of multiple genotypes and provides the basis for the development of a multivalent vaccine against genetically diverse NoV infections.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115038"},"PeriodicalIF":2.2,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plate centrifugation enhances the efficiency of polyethylenimine-based transfection and lentiviral infection","authors":"Shaozhe Yang ,&nbsp;Qingwei Zhang ,&nbsp;Yuan Zhuang ,&nbsp;Junfeng Li ,&nbsp;Xiuhong Fu","doi":"10.1016/j.jviromet.2024.115039","DOIUrl":"10.1016/j.jviromet.2024.115039","url":null,"abstract":"<div><h3>Purpose</h3><div>To propose an efficient, reproducible, and consistent transgenic technology based on plate centrifugation, which is particularly useful for polyethylenimine (PEI) transfection and lentiviral infection.</div></div><div><h3>Methods</h3><div>We optimized multiple factors that could contribute to transfection efficiency, such as the dosage of the PEI or DNA, the working solution buffer used for diluting the PEI or DNA, the incubation time for the PEI/DNA complexes, and the transfection time.</div></div><div><h3>Results</h3><div>Plate centrifugation led to a 5.46-fold increase in the transfection efficiency of PEI-based transfection while maintaining the cell survival rate. Moreover, the average copy number of viral genes in each genome increased 4.96-fold with plate centrifugation. Plate centrifugation alters the spatial arrangement of the PEI/DNA complexes or lentiviruses, increasing the chances of these complexes or viruses coming into contact with target cells, ultimately resulting in improved transfection or infection efficiency.</div></div><div><h3>Conclusions</h3><div>We present a protocol based on plate centrifugation for transfection or lentiviral infection that is suitable for genetic modification of primary cells or stem cells.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115039"},"PeriodicalIF":2.2,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142365665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A method for producing protease pS273R of the African swine fever virus","authors":"Danil S. Kalinin ,&nbsp;Sergey G. Mayorov ,&nbsp;Marina Yu. Zemskova ,&nbsp;Oleg R. Latypov ,&nbsp;Michael G. Shlyapnikov ,&nbsp;Maria A. Gorshkova ,&nbsp;Eva N. Titova ,&nbsp;Natalia N. Vlasova ,&nbsp;Alexey V. Lipkin ,&nbsp;Alexey N. Fedorov ,&nbsp;Igor E. Granovsky","doi":"10.1016/j.jviromet.2024.115037","DOIUrl":"10.1016/j.jviromet.2024.115037","url":null,"abstract":"<div><div>The pS273R protease of the African swine fever virus (ASFV) is responsible for the processing of the viral polyproteins pp220 and pp62, precursors of the internal capsid of the virus. The protease is essential for a productive viral infection and is an attractive target for antiviral therapy. This work presents a method for the production of pS273R in <em>E. coli</em> cells by fusing the protease with the SlyD chaperone. The chimeric protein pS273R protease, during expression, is formed in a soluble form possessing enzymatic activity. Subsequently, pS273R separates from SlyD through autocatalytic cleavage at the TEV protease site <em>in vivo</em>. This work devised a straightforward protocol for chromatographic purification, resulting in the production of a highly purified viral protease. Additionally, we suggest using a fluorescence method to assess the activity of pS273R. This method is predicated on a shift in the chimeric protein thioredoxin-EGFP's electrophoretic mobility following its protease cleavage. It was shown that thioredoxin-EGFP substrate is effectively and selectively cleaved by the pS273R protease, even in complex protein mixtures such as mammalian cell lysates.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115037"},"PeriodicalIF":2.2,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142328095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a quantitative PCR assay for detection of Sulawesi tortoise adenovirus","authors":"Zachary C. Ready ,&nbsp;Laura Adamovicz ,&nbsp;Maris Daleo ,&nbsp;Amber Simmons ,&nbsp;Matthew C. Allender","doi":"10.1016/j.jviromet.2024.115033","DOIUrl":"10.1016/j.jviromet.2024.115033","url":null,"abstract":"<div><div>In 2007, a mortality event involving over 100 Sulawesi tortoises (<em>Indotestudo forsteni</em>), two Impressed tortoises (<em>Manouria impress</em>) and a critically endangered Burmese star tortoise (<em>Geochelone platynota</em>) was attributed to Sulawesi tortoise adenovirus (STADV; genus <em>Siadenovirus</em>). We developed a TaqMan quantitative PCR assay targeting the DNA polymerase gene of STADV for use in clinical diagnosis and epidemiologic surveillance. This assay failed to amplify five closely-related chelonian adenoviruses, indicating high analytical specificity. The assay performed with high efficiency (slope = −3.337; R² = 0.999) and high inter- and intra-assay repeatability (coefficient of variation &lt;1.36 % at all standard curve dilutions). Dynamic range included 1.00 × 10⁷ to 1.00 × 10¹ target copies per reaction and limit of detection was 10¹ target copies per reaction, though 10⁰ target copies per reaction were intermittently detected. This qPCR assay provides a valuable diagnostic tool for characterization of STADV epidemiology, including potential identification of the North American reservoir host.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115033"},"PeriodicalIF":2.2,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142308002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reducing amplification cycles to improve the coverage of influenza A virus genome sequencing in heterosubtypic co-infection","authors":"Yijie Zhang ,&nbsp;Wenhua Kong ,&nbsp;Yixuan Wu ,&nbsp;Zhi Chen ,&nbsp;Xiang Zhao ,&nbsp;Manqing Liu","doi":"10.1016/j.jviromet.2024.115036","DOIUrl":"10.1016/j.jviromet.2024.115036","url":null,"abstract":"<div><div>This study delineates the enhancement of a Reverse Transcription Polymerase Chain Reaction (RT-PCR) method for the amplification of the complete genome of the influenza A virus during heterosubtypic co-infection, relying on the amplification of intact gene segments. The precision of the method was assessed using all amplicons, which underwent both capillary electrophoresis and DNA sequencing. Five samples featuring co-infection of Influenza A viruses with H1N1 and H3N2 subtypes were evaluated. The improved strategy successfully amplified all eight segments of H3N2 strains in four samples, and the entire genome of H1N1 strains in three samples.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115036"},"PeriodicalIF":2.2,"publicationDate":"2024-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142290212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Performance of an in-house multiplex PCR assay for HIV-1 drug resistance testing – A cheaper alternative","authors":"Tumelo L. Fortuin ,&nbsp;Paballo Nkone ,&nbsp;Allison Glass ,&nbsp;Raquel Viana ,&nbsp;Keitumetse Moeng ,&nbsp;Shayne Loubser ,&nbsp;Caroline T. Tiemessen ,&nbsp;Simnikiwe H. Mayaphi","doi":"10.1016/j.jviromet.2024.115034","DOIUrl":"10.1016/j.jviromet.2024.115034","url":null,"abstract":"<div><h3>Background</h3><div>Currently, most HIV drug resistance PCR assays amplify the protease-reverse transcriptase (PR-RT) fragment separately from the integrase (IN) fragment. The aim of this study was to develop a multiplex PCR assay that simultaneously amplifies PR-RT and IN fragments for HIV-1 drug-resistance testing.</div></div><div><h3>Methods</h3><div>The in-house multiplex PCR assay was evaluated on extracted total nucleic acids obtained from the National Health Laboratory Service (NHLS) and Lancet laboratories. Sanger sequencing was performed on amplicons, and HIV-1 drug-resistance mutations (DRMs) were assessed using HIV Stanford drug resistance database.</div></div><div><h3>Results</h3><div>This study tested 59 patient samples with known HIV-1 viral load and DRM results; 41 from Lancet and 18 from NHLS. In-house multiplex PCR assay detected one or both fragments in most samples but had higher sensitivity for detection of IN fragment (93.2 %) compared to PR-RT fragment (83.1 %). There was 100 % concordance between Lancet assay versus in-house assay sequence data for IN DRMs, but lower concordance with PR-RT (87.0 %). The in-house multiplex PCR assay’s precision and reproducibility analysis showed ≥99.9 % sequence similarity and yielded similar DRM results for both PR-RT and IN fragments.</div></div><div><h3>Conclusions</h3><div>The in-house multiplex PCR assay demonstrated satisfactory performance and higher sensitivity for IN fragment amplification. This could be a cost-effective method for HIV-1 drug resistance testing as both PR-RT and IN fragments are successfully amplified in one reaction in most samples.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115034"},"PeriodicalIF":2.2,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0166093424001587/pdfft?md5=1d8773b905f46d2f79c1de4d12dac17f&pid=1-s2.0-S0166093424001587-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142290211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid detection of bat coronaviruses from fecal samples using loop-mediated isothermal amplification assay in the field","authors":"Undarmaa Tsengel ,&nbsp;Tzong-Yuan Wu ,&nbsp;Yi-Ning Chen","doi":"10.1016/j.jviromet.2024.115035","DOIUrl":"10.1016/j.jviromet.2024.115035","url":null,"abstract":"<div><p>The global impact of the COVID-19 pandemic has emphasized the critical need for effective viral diagnostics. Although polymerase chain reaction (PCR) is a well-established nucleotide amplification technique, its limitations, such as the need for expensive equipment and skilled technicians, have led to the exploration of alternative methods, including loop-mediated isothermal amplification (LAMP). Bats, as a crucial natural reservoir of coronaviruses (CoVs), particularly <em>Scotophilus</em> bat coronavirus 512 (<em>Scotophilus</em> bat-CoV 512) prevalent among Taiwan’s bat population, are the focus of this study. We aimed to detect <em>Scotophilus</em> bat-CoV 512 from bats in field conditions using loop-mediated isothermal amplification (LAMP) assay for on-site detection. Therefore, our study delves into the specificity of the LAMP reaction, emphasizing the careful design of primers to prevent false positive results. A cross reactivity and primer specificity test involving seven different microorganisms, including closely related bat CoVs and two bacterial species typically found in feces, revealed that the LAMP assay uniquely detected <em>Scotophilus</em> bat-CoV 512. The developed colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was optimized for the primers targeting nucleocapsid (N) gene, and the sensitivity test revealed a detection limit of 2.4 × 10³ copies/µL. Our findings indicate the potential of the RT-LAMP assay for on-site detection in the field and subsequent laboratory analysis for comprehensive sampling and further research on bat CoV isolation. The surveillance and monitoring of bat CoVs contribute substantially to mitigating human threats, particularly concerning the emergence of new pandemic variants.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115035"},"PeriodicalIF":2.2,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142271062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}