Journal of virological methods最新文献

{"title":"Performance of an envelope glycoprotein-based multiplex immunoassay for Ebola virus antibody detection in a cohort of Ebola virus disease survivors","authors":"McKenna D. Roe ,&nbsp;Grace Hood ,&nbsp;Spencer L. Sterling ,&nbsp;Lianying Yan ,&nbsp;Joseph Akoi Boré ,&nbsp;Tom Tipton ,&nbsp;Craig Thompson ,&nbsp;Miles W. Carroll ,&nbsp;Eric D. Laing","doi":"10.1016/j.jviromet.2024.115057","DOIUrl":"10.1016/j.jviromet.2024.115057","url":null,"abstract":"<div><div>Serological surveillance in animal and human hosts can be a cost-effective strategy for orthoebolavirus detection, but is challenged by accurate estimates of seroprevalence, potential pauci-symptomatic disease presentation, and antigenic cross-reactivity. Here, we describe the use of an envelope glycoprotein (GP)-based multiplex microsphere immunoassay, consisting of nine filovirus GP antigens for the detection of anti-Ebola virus (EBOV) antibodies in a well-characterized cohort of Guinean Ebola virus disease (EVD) survivors and contacts from the 2013 – 2016 West African EVD outbreak. We examined sensitivity and specificity for the detection of anti-EBOV antibodies by GP expressed as recombinant trimeric ectodomains, yielding an assay performance of 95.96 % sensitivity and 98.61 % specificity. Additionally, agreement between the multiplex test and a whole virus ELISA and virus neutralization test showed strong correlations. The results demonstrate that this filovirus multiplex test is a sensitive tool for high-throughput serosurveillance</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115057"},"PeriodicalIF":2.2,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142503031","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":"Preliminary survey of three mosquito-borne viruses using a self-established multiplex RT-qPCR assay in Chinese blood donors","authors":"Xipeng Yan ,&nbsp;Xinwei Wang ,&nbsp;Jinlian Li ,&nbsp;Bin Li ,&nbsp;Baoren He ,&nbsp;Linbin Huang ,&nbsp;Jingheng Liang ,&nbsp;Min Xu ,&nbsp;Limin Chen","doi":"10.1016/j.jviromet.2024.115055","DOIUrl":"10.1016/j.jviromet.2024.115055","url":null,"abstract":"<div><h3>Background</h3><div>Mosquito-borne pathogens pose a significant threat to both human health and blood safety. The primary mosquito-borne viruses that present this threat are Zika virus, Chikungunya virus, and Dengue virus. At present, there are limited efficacious vaccines or therapeutic drugs for the prevention and treatment of these viral infections. Blood donors can remain asymptomatically infected and unfortunately, screening for these three viruses in Chinese blood donors are not mandatory, leaving the residual risk to transfusion recipients uncertain. Objective: To address this, we developed a single-tube multiplex RT-qPCR assay for ZCD detection and was preliminarily employed to screen a total of 10,566 blood donations in Nanning Blood Center in order to assess the prevalence risk of these pathogens in blood donors. Results: None of the blood samples was reactive for ZCD by nucleic acid test (NAT). One out of 173 donations (1/173, 0.58 %) was IgG positive for ZIKV and 14 (14/173, 8.4 %) were IgG positive for DENV. None of these 173 donations was IgG positive for Chikungunya virus. These findings suggest that the prevalence of ZCD infection in blood donors in Nanning is very low although past DENV infection (IgG positive) was relatively common. Conclusion: A single-tube multiplex RT-qPCR assay for simultaneous detection of ZCD viruses was successfully established and applied for screening in blood donors. The residual risk of ZCD infection through transfusion is currently low in Nanning, China. The NAT assay for ZCD will serve as a technical reserve in response to future epidemic or pandemic of mosquito-borne pathogens.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115055"},"PeriodicalIF":2.2,"publicationDate":"2024-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142503032","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":"Adjusting susceptibilities of C57BL/6 mice to orthoflaviviruses for evaluation of antiviral drugs by altering the levels of interferon alpha/beta receptor function","authors":"John D. Morrey,&nbsp;Venkatraman Siddharthan","doi":"10.1016/j.jviromet.2024.115053","DOIUrl":"10.1016/j.jviromet.2024.115053","url":null,"abstract":"<div><div>The purpose of this study was to optimize the infectivity of four different orthoflaviviruses in mice for evaluating antiviral drugs by using wild-type mice with intact interferon responses, type 1 interferon alpha/beta receptor knockout mice, or by injecting wild type C57BL/6 mice with varying doses of anti-type 1 interferon receptor antibody (MAR1-5A3) to optimize the infectivity and lethality. West Nile virus productively infected wild-type C57BL/6 mice to cause lethality, whereas Usutu virus required a complete absence of type 1 interferon receptor function. Deer tick virus (lineage 2 Powassan virus) and Japanese encephalitis viruses required a dampening of type 1 interferon responses by adjusting the doses of MAR1-5A3 antibody injections. Challenge dose-responsive mortality, weight loss, and viral titers of these two viruses were observed if the type 1 interferon responses were dampened with MAR1-5A3. Conversely, without MAR1-5A3 injections, these disease phenotypes were not viral challenge dose-responsive. From these different interferon-responsive models, the appropriate lethality was identified to determine that 7-deaza-2’-C-methyladenosine has high efficacy for West Nile and Usutu viruses, and low efficacy for deer tick and Japanese encephalitis viruses.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115053"},"PeriodicalIF":2.2,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468816","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":"One-step TaqMan® RT-qPCR detection of sugar beet-infecting poleroviruses in Myzus persicae from yellow water pan traps opens up new possibilities for early risk assessment of virus yellows disease","authors":"Simon Borgolte ,&nbsp;Wulf Menzel ,&nbsp;Mark Varrelmann","doi":"10.1016/j.jviromet.2024.115052","DOIUrl":"10.1016/j.jviromet.2024.115052","url":null,"abstract":"<div><div>Virus yellows disease (VY) is a major threat to sugar beet production in Europe. Beet chlorosis virus (BChV) and beet mild yellowing virus (BMYV) are of particular economic importance and are both persistently transmitted by the aphid vector <em>Myzus persicae</em>. As part of integrated pest management strategies, <em>M. persicae</em> influx into sugar beet fields is recorded weekly using yellow water pan traps. To date, only ELISA and RT-PCR assays have been described for BChV and BMYV detection in individual aphids. In this study, we describe for the first time two one-step TaqMan® RT-qPCR assays designed for the specific detection of BChV and BMYV in <em>M. persicae</em> after 7d incubation in water pan trap medium. Both viruses were reproducibly detected in individual aphids. After 7d incubation in trap medium, both viruses were reproducibly detected in individual aphids, as well as in one viruliferous aphid in a pool of 99 non-viruliferous aphids. Significant correlations can be shown between different mixing ratios of viruliferous to non-viruliferous aphids and Ct values of total RNA templates, allowing the percentage of viruliferous aphids in yellow water pan traps to be estimated using a standard curve. The described methodology provides a high sensitivity combined with a high sample throughput and can be used, after evaluation in the field, for practical monitoring, risk modelling and development of decision support systems for VY.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115052"},"PeriodicalIF":2.2,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468818","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":"Analysis of the antigenic and immunogenic properties of the native rabies virus glycoprotein purified by Lens culinaris lectin affinity chromatography","authors":"Cíntia Pinto da Silva,&nbsp;Talita Gonçalves Aires de Queiroz,&nbsp;Keila Iamamoto Nogi,&nbsp;Iana Suly Santos Katz,&nbsp;Fernanda Guedes,&nbsp;Elaine Raniero Fernandes,&nbsp;Karina Ribeiro Silva,&nbsp;Sandriana Ramos Silva","doi":"10.1016/j.jviromet.2024.115044","DOIUrl":"10.1016/j.jviromet.2024.115044","url":null,"abstract":"<div><div>Rabies virus glycoprotein (RABV-G) is responsible for the recognition of specific cell surface receptors and induces the production of neutralizing antibodies (VNA). Since RABV-G is a glycoprotein, this work aimed to evaluate <em>Lens culinaris</em> (LCA) chromatography as a simple and effective purification method. The purity and identification of the protein obtained were analyzed by SDS-PAGE, ELISA and lectin-binding assay. The antigenic properties of the purified RABV-G were evaluated by direct ELISA using human serum samples from individuals who had received rabies pre-exposure vaccination. For the immunogenicity study, Swiss Webster mice were immunized with purified RABV-G and the specific antibodies were measured by direct ELISA and RFFIT. As results, it was observed that the purified protein reveled a molecular mass of 55 kDa and the presence of carbohydrate; additionally, it was recognized by anti-rabies virus glycoprotein monoclonal antibody. Purified RABV-G induced high VNA titers (&gt;50.0 IU/ml) <em>in vivo</em>, as detected by RFFIT, as well as RABV-G specific IgG1 (0.8 mean OD±SD) and IgG2a (0.3 mean OD±SD) antibodies, with a predominance of IgG1 (p&lt; 0.001). In addition, it was observed that RABV-G was efficient in selectively detecting anti- RABV-G IgG in the sera of vaccinated individuals compared to the negative control. Therefore, LCA chromatography was efficient in preserving the native properties of RABV-G that are essential in inducing an adequate humoral immune response. In addition, the purified RABV-G presented analytical potential as an ELISA reagent.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115044"},"PeriodicalIF":2.2,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468817","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":"Infectivity titers and aggregation states of intracellular and extracellular nervous necrosis virus in cell lines with cytolytic and persistent infections","authors":"Han Sol Lee ,&nbsp;Toyohiko Nishizawa","doi":"10.1016/j.jviromet.2024.115043","DOIUrl":"10.1016/j.jviromet.2024.115043","url":null,"abstract":"<div><div>Nervous necrosis virus (NNV) in the genus <em>Betanodavirus</em> (<em>Nodaviridae</em>) is highly lethal to a wide range of fish species. Although striped snakehead (SSN-1) cell lines have been widely used for culturing NNV, cell lines persistently infected (PI) with NNV have only recently been established. This study investigated the infectivity titers of intracellular and extracellular NNV and the associated aggregation states. The intracellular NNV infectious doses were higher than those of extracellular NNV, irrespective of the cell lines. In SSN-1 cells, the intracellular-to-extracellular-NNV ratio was approximately 50–60-fold on days 1 and 2 after NNV infection, although it decreased following the onset of the cytopathic effect (CPE), reaching 3.5-fold on day 4. In the PI-cell lines, both the intracellular and extracellular NNV were in a nearly monomeric state. While the extracellular NNV were in a monomeric state in the SSN-1 cells, more than 92 % of the intracellular virus were in an aggregated state. When the NNV accumulated intracellularly at a median tissue culture infectious dose (TCID₅₀)/cell of 10⁴ to 10^4.5, SSN-1 cells appeared to exhibit CPE and eventually died. We believe that the aggregates of intracellularly accumulated NNV particles may be related to the cellular CPE onset and/or cell death.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"330 ","pages":"Article 115043"},"PeriodicalIF":2.2,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406579","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":"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}