Journal of virological methods最新文献_第4页

Experimental arboviral infection of mosquito larvae: A novel approach for vector competence studies 蚊子幼虫的虫媒病毒实验感染：病媒能力研究的新方法。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-07 DOI: 10.1016/j.jviromet.2024.115061

Christin Körsten, Mandy Schäfer

{"title":"Experimental arboviral infection of mosquito larvae: A novel approach for vector competence studies","authors":"Christin Körsten, Mandy Schäfer","doi":"10.1016/j.jviromet.2024.115061","DOIUrl":"10.1016/j.jviromet.2024.115061","url":null,"abstract":"<div><div>Vector competence studies in mosquitoes present valuable opportunities to explore arboviral transmission and virus-vector interactions. However, oral infection studies in mosquitoes can be challenging. An alternative approach is to infect mosquitoes during their aquatic larval stage, resulting in the emergence of infected adults. To investigate the potential of this method, <em>Culex pipiens</em> biotype <em>molestus</em> larvae were infected with Usutu virus (USUV, <em>Orthoflavivirus usutuense</em>). For this purpose, larvae were exposed to USUV-infected mammalian and mosquito cell cultures for 24 h before being reared to adults. Subsequent analysis via RT-qPCR revealed that the <em>Culex</em> larvae successfully acquired USUV from the infected cells and exhibited high susceptibility to infection. Immediately after emergence, 32.10 % (26/81) of male and 41.03 % (16/39) of female mosquitoes tested positive for USUV RNA. Notably, females that were incubated for 15 days post-emergence demonstrated even higher infection rates, reaching 100.00 % (23/23). In addition, viral RNA and infectious particles were detected in some saliva samples, indicating the potential for transmission. This experimental infection of mosquito larvae thus offers the opportunity to produce infected adult mosquitoes for studies enhancing our understanding of virus-vector interactions, co-infections, and transmission routes. Such research contributes to better public health strategies addressing arboviral diseases.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115061"},"PeriodicalIF":2.2,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142622906","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}

引用次数: 0

Development of a concentration method for simple and sensitive detection of SARS-CoV-2 in saliva using a magnetic nanoparticle kit 利用磁性纳米粒子试剂盒开发唾液中 SARS-CoV-2 简单灵敏检测浓度法。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-01 DOI: 10.1016/j.jviromet.2024.115059

Yasuko Yamazaki , Riku Tanaka , Gladys Castillo , Adrian Miki C. Macalanda , Melbourne R. Talactac , Wataru Yamazaki

{"title":"Development of a concentration method for simple and sensitive detection of SARS-CoV-2 in saliva using a magnetic nanoparticle kit","authors":"Yasuko Yamazaki , Riku Tanaka , Gladys Castillo , Adrian Miki C. Macalanda , Melbourne R. Talactac , Wataru Yamazaki","doi":"10.1016/j.jviromet.2024.115059","DOIUrl":"10.1016/j.jviromet.2024.115059","url":null,"abstract":"<div><h3>Background</h3><div>When diagnosing viral infections in humans and animals, the presence of virus in a sample in trace amounts that are below the analytical sensitivity of the detection system may cause false negative results and inaccurate diagnosis. We previously reported the development of a simple virion concentration technique using 12 ml large-volume samples that can dramatically improve diagnostic sensitivity by increasing analytical sensitivity by 100-fold over conventional methods. The present study was conducted to further improve the simplicity and versatility of this method. We constructed a simple and highly sensitive method for the detection of SARS-CoV-2 in human saliva after concentration using a magnetic nanoparticle conjugated with polyethylene glycol (PEG).</div></div><div><h3>Results</h3><div>Performance of the method was evaluated by comparing a combination of automated nucleic acid extraction and RT-qPCR or triplex RT-LAM detection in a spiked sample of 20 ml saliva collected from healthy humans. The method theoretically achieved 300-fold concentration of spiked SARS-CoV-2 in saliva, enabling 10- to 1000-fold higher analytical sensitivity for detection compared to conventional RNA extraction methods.</div></div><div><h3>Conclusions</h3><div>This newly developed method allows for easy and reliable concentration of the virion in less than 60 min, improving the analytical sensitivity of the SARS-CoV-2 test. Further, the method allows for easy and reliable enrichment of the virus in less than 60 min, improving the analytical sensitivity of the SARS-CoV-2 test. This method is easily used for highly sensitive virus detection from a variety of human oral fluid samples and may also be applied to rapid and labor-saving screening tests by pooling a large number of samples.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115059"},"PeriodicalIF":2.2,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566994","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}

引用次数: 0

Indirect ELISA for analysis of malignant catarrhal fever virus-specific antibodies in a range of species 用于分析各种物种中恶性卡他热病毒特异性抗体的间接酶联免疫吸附试验。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-31 DOI: 10.1016/j.jviromet.2024.115060

George C. Russell, Ann Percival, Dawn M. Grant

引用次数: 0

Effects of chemokine-binding protein in visceral ovine aphthae on immune regulation response 内脏鹅口疮中的趋化因子结合蛋白对免疫调节反应的影响

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-30 DOI: 10.1016/j.jviromet.2024.115058

Fu Guowen , Li Rong , Zhang Ruixue, Zeng Qin, Yuan Jiarui, Liu Yabo, Fan Yueyuan

{"title":"Effects of chemokine-binding protein in visceral ovine aphthae on immune regulation response","authors":"Fu Guowen , Li Rong , Zhang Ruixue, Zeng Qin, Yuan Jiarui, Liu Yabo, Fan Yueyuan","doi":"10.1016/j.jviromet.2024.115058","DOIUrl":"10.1016/j.jviromet.2024.115058","url":null,"abstract":"<div><div>ORFV of the family poxvirus，which produces a pustular dermatitis both in humans and animals.,Previous studies have found an fatal case caused by the infection of ORFV in the viscera. However, the mechanisms of ORFV how to infect the viscera remain unknown. Our sequencing results revealed that the CBP of the visceral infection strain lacked a 24-base pair segment at position 217 comparison to the oral infection strain. Subsequently, we successfully packaged the recombinant adenoviruses pAd-CBP-K and pAd-CBP-N in HEK-293A cells and carried it to infect lymphocytes. RT-PCR analysis showed that the CBP protein was expressed in lymphocytes, and pAd-CBP-N group exhibited a significantly higher CBP expression level compared to the pAd-CBP-K group. At 4, 8, and 12 hours post-infection, both pAd-CBP-K and pAd-CBP-N were found to downregulate the expression of MIP-1 and CCL-5 in the supernatant of lymphocytes. However, the expression of IL-2, IL-6, IL-8, IL-12, INF-γ, and TNF-α showed a significant up-regulation. Furthermore, the inflammatory factors relative expression levels of IL-2, IL-6, IL-8, IL-8, IL-12, IFN-γ and TNF-α were significantly up-regulated in the both group. Interestingly, a significant increase in the expression of IL-6, IL-8 and TNF-α were detected in the pAd-CBP-N group at both 8 and 12 hours compared to pAd-CBP-N. Taken together, these findings showed that CBP can regulate the expression of free chemokines and activate the expression of inflammatory factors, and provide a basis for follow-up research.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115058"},"PeriodicalIF":2.2,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564387","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}

引用次数: 0

Performance of an envelope glycoprotein-based multiplex immunoassay for Ebola virus antibody detection in a cohort of Ebola virus disease survivors 基于包膜糖蛋白的多重免疫测定在一组埃博拉病毒病幸存者中检测埃博拉病毒抗体的性能。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-24 DOI: 10.1016/j.jviromet.2024.115057

McKenna D. Roe , Grace Hood , Spencer L. Sterling , Lianying Yan , Joseph Akoi Boré , Tom Tipton , Craig Thompson , Miles W. Carroll , Eric D. Laing

{"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 , Grace Hood , Spencer L. Sterling , Lianying Yan , Joseph Akoi Boré , Tom Tipton , Craig Thompson , Miles W. Carroll , 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}

引用次数: 0

Preliminary survey of three mosquito-borne viruses using a self-established multiplex RT-qPCR assay in Chinese blood donors 利用自建的多重 RT-qPCR 分析法对中国献血者中的三种蚊媒病毒进行初步调查。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-20 DOI: 10.1016/j.jviromet.2024.115055

Xipeng Yan , Xinwei Wang , Jinlian Li , Bin Li , Baoren He , Linbin Huang , Jingheng Liang , Min Xu , Limin Chen

{"title":"Preliminary survey of three mosquito-borne viruses using a self-established multiplex RT-qPCR assay in Chinese blood donors","authors":"Xipeng Yan , Xinwei Wang , Jinlian Li , Bin Li , Baoren He , Linbin Huang , Jingheng Liang , Min Xu , 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}

引用次数: 0

Adjusting susceptibilities of C57BL/6 mice to orthoflaviviruses for evaluation of antiviral drugs by altering the levels of interferon alpha/beta receptor function 通过改变干扰素α/β受体功能水平，调整C57BL/6小鼠对正黄病毒的敏感性，以评估抗病毒药物。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-18 DOI: 10.1016/j.jviromet.2024.115053

John D. Morrey, Venkatraman Siddharthan

{"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, 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}

引用次数: 0

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 一步式 TaqMan® RT-qPCR 检测黄水盘诱捕器中甜菜瘤蚜感染的多角体病毒，为病毒黄化病的早期风险评估提供了新的可能性。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-16 DOI: 10.1016/j.jviromet.2024.115052

Simon Borgolte , Wulf Menzel , Mark Varrelmann

{"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 , Wulf Menzel , 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}

引用次数: 0

Analysis of the antigenic and immunogenic properties of the native rabies virus glycoprotein purified by Lens culinaris lectin affinity chromatography 用 Lens culinaris 凝集素亲和层析法纯化的原生狂犬病病毒糖蛋白的抗原性和免疫原性分析。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-14 DOI: 10.1016/j.jviromet.2024.115044

Cíntia Pinto da Silva, Talita Gonçalves Aires de Queiroz, Keila Iamamoto Nogi, Iana Suly Santos Katz, Fernanda Guedes, Elaine Raniero Fernandes, Karina Ribeiro Silva, Sandriana Ramos Silva

{"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, Talita Gonçalves Aires de Queiroz, Keila Iamamoto Nogi, Iana Suly Santos Katz, Fernanda Guedes, Elaine Raniero Fernandes, Karina Ribeiro Silva, 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 (>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< 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}

引用次数: 0

Infectivity titers and aggregation states of intracellular and extracellular nervous necrosis virus in cell lines with cytolytic and persistent infections 细胞溶解性和持续性感染细胞系中细胞内和细胞外神经坏死病毒的感染滴度和聚集状态。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-10-10 DOI: 10.1016/j.jviromet.2024.115043

Han Sol Lee , Toyohiko Nishizawa

{"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 , 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 (TCID<sub>50</sub>)/cell of 10<sup>4</sup> to 10<sup>4.5</sup>, 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}

引用次数: 0