Journal of virological methods最新文献_第4页

The optimisation and application of a novel BaseScope™ RNA-ISH assay for the detection of foot-and-mouth disease virus in carrier African buffalo (Syncerus caffer) from South Africa 新型BaseScope™RNA-ISH检测方法的优化和应用，用于检测南非非洲水牛（Syncerus caffer）携带的口蹄疫病毒

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-08-05 DOI: 10.1016/j.jviromet.2025.115237

Alischa Henning , Lieza Odendaal , Angelika Loots , Melvyn Quan

{"title":"The optimisation and application of a novel BaseScope™ RNA-ISH assay for the detection of foot-and-mouth disease virus in carrier African buffalo (Syncerus caffer) from South Africa","authors":"Alischa Henning , Lieza Odendaal , Angelika Loots , Melvyn Quan","doi":"10.1016/j.jviromet.2025.115237","DOIUrl":"10.1016/j.jviromet.2025.115237","url":null,"abstract":"<div><div>The detection of foot-and-mouth disease is limited to BSL-3 laboratories and its detection in carrier animals require increased test sensitivity. <em>In-situ</em>-hybridisation utilises the propensity of a labelled single-stranded sequence of DNA or RNA to anneal to a complementary target. It can be performed on formalin-fixed tissues and with some of the most recent advances, show an increased test sensitivity. BaseScope™ incorporates an additional signal amplification step, which makes it possible to detect RNA splicing variants, point mutations, small insertions or deletions, and short RNA targets (50–300 nucleotides). This study aimed to adjust and optimise the BaseScope™ assay to detect foot-and-mouth disease virus in a novel, carrier wildlife species, i.e., buffalo. Specific steps were adjusted to attempt to address some of the rigidity involved in the workflow. However, none of the in-house reagents or equipment attempted as an alternative to the original and prescribed workflow was successful. This demonstrates the fastidious nature of this diagnostic modality and the synergistic characteristics of a commercial assay. However, keeping tissues in formalin for up to 7 days and storing cut sections for up to 3 months did not have a negative impact on the results. This further demonstrated the reliability of BaseScope™.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115237"},"PeriodicalIF":1.6,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144781323","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

Limitations and use of the Morpheus-V5 dual reporter virus in assessing interventions that target HIV latency Morpheus-V5双重报告病毒在评估针对HIV潜伏期的干预措施中的局限性和使用

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-08-05 DOI: 10.1016/j.jviromet.2025.115236

Kiho Tanaka , Youry Kim , Jesslyn Ong , Carolin Tumpach , Ajantha Rhodes , Hannah AD King , Anna C. Hearps , Michael Roche , Sharon R. Lewin

{"title":"Limitations and use of the Morpheus-V5 dual reporter virus in assessing interventions that target HIV latency","authors":"Kiho Tanaka , Youry Kim , Jesslyn Ong , Carolin Tumpach , Ajantha Rhodes , Hannah AD King , Anna C. Hearps , Michael Roche , Sharon R. Lewin","doi":"10.1016/j.jviromet.2025.115236","DOIUrl":"10.1016/j.jviromet.2025.115236","url":null,"abstract":"<div><div>HIV can persist indefinitely in latently infected CD4 + T-cells as an integrated provirus with limited or no viral transcription and expression of viral proteins. We further characterised a recently described dual reporter virus, Morpheus-V5, that expresses murine heat-stable antigen and mCherry in productively infected cells (which is HIV LTR dependent) and V5 and Nerve growth factor receptor (NGFR) in latently infected cells (which is HIV LTR independent). We demonstrated successful infection of resting and activated CD4 + T-cells using Morpheus-V5 pseudotyped with either X4, R5 or dual tropic envelope proteins. We also showed that expression of NGFR (a transmembrane protein) enriched for infected cells (that contained HIV DNA) with inducible virus, however uninfected cells also expressed NGFR as a result of NGFR incorporation into virion preparations. Following treatment of CD4 + T-cells infected with Morpheus-V5 with latency reversal agents, we demonstrated an increase in the percentage of cells expressing mCherry and a decrease in the percentage of cells expressing NGFR. In addition, using this model, we showed that latent and productively infected cells had different levels of sensitivity to pro-apoptotic compounds. The Morpheus-V5 dual reporter virus has some limitations and overestimates the number of latently infected cells, but is a useful tool to investigate interventions that disrupt HIV latency.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115236"},"PeriodicalIF":1.6,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144775761","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

Utilizing multiplex reverse transcription-multiple cross displacement amplification-lateral flow biosensor technology for detecting H1N1, H3N2 and H7N9 influenza A virus subtypes 利用多重逆转录-多重交叉位移扩增-侧流生物传感器技术检测H1N1、H3N2和H7N9甲型流感病毒亚型

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-08-04 DOI: 10.1016/j.jviromet.2025.115235

Pengfei Wang , Linlin Yan , Jing Wang , Shoukui Hu , Fan Zhao

{"title":"Utilizing multiplex reverse transcription-multiple cross displacement amplification-lateral flow biosensor technology for detecting H1N1, H3N2 and H7N9 influenza A virus subtypes","authors":"Pengfei Wang , Linlin Yan , Jing Wang , Shoukui Hu , Fan Zhao","doi":"10.1016/j.jviromet.2025.115235","DOIUrl":"10.1016/j.jviromet.2025.115235","url":null,"abstract":"<div><div>Respiratory infections caused by influenza A viruses (IAVs) pose a global threat annually, leading to significant mortality in severe cases. IAVs are classified into various subtypes, and a combination of multiplex reverse transcription-multiple cross displacement amplification (mRT-MCDA) and lateral flow biosensor (LFB) has been developed to detect three subtypes (H1N1, H3N2, and H7N9) that have been frequently observed in recent years. This technology simultaneously reverse transcribes and amplifies two target genes, hemagglutinin (HA) and neuraminidase (NA), at a constant temperature of 63 °C in just 40 min. The final results can be read using a dual-channel LFB. In practice, the entire process—from sample collection, RNA extraction, and mRT-MCDA amplification to result interpretation—can be completed in less than 1 h. This method is named \"multiplex reverse transcription-multiple cross displacement amplification-lateral flow biosensor\" (IAVs-mRT-MCDA-LFB) technology. The IAVs-mRT-MCDA-LFB technology demonstrated higher sensitivity compared to conventional reverse transcription-polymerase chain reaction (RT-PCR) and showed no cross-reactivity with other genes present in the samples. Therefore, the IAVs-mRT-MCDA-LFB technique developed in this study is a highly valuable molecular diagnostic tool for detecting IAV-subtypes due to its simplicity, time efficiency, cost-effectiveness, and high specificity and sensitivity.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115235"},"PeriodicalIF":1.6,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144771821","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

Characterization of Goatpox virus major intracellular mature virion proteins A27L and P32 expressed by baculovirus system: Superior to Escherichia coli 杆状病毒系统表达羊痘病毒主要细胞内成熟病毒粒子蛋白A27L和P32的特性：优于大肠杆菌

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-07-30 DOI: 10.1016/j.jviromet.2025.115234

Anand Kushwaha, Poulinlu Golmei, Amit Kumar, B. Mondal, Gnanavel Venkatesan

{"title":"Characterization of Goatpox virus major intracellular mature virion proteins A27L and P32 expressed by baculovirus system: Superior to Escherichia coli","authors":"Anand Kushwaha, Poulinlu Golmei, Amit Kumar, B. Mondal, Gnanavel Venkatesan","doi":"10.1016/j.jviromet.2025.115234","DOIUrl":"10.1016/j.jviromet.2025.115234","url":null,"abstract":"<div><div>Baculovirus expression system has become the most widely used eukaryotic system for recombinant protein production due to its ability to perform post-translational modifications and produce proteins with native-like conformation and antigenicity. In this study, two major immunodominant intracellular mature virion genes of Goatpox virus were amplified into three constructs: A27L, full-length P32, and truncated P32 (tr-P32), respectively. The constructs were cloned into the pFastBac HTA vector and expressed in <em>Trichoplusiani</em> (TN5) insect cells using the baculovirus expression system. Recombinant A27L was purified under native conditions, however full-length P32 and tr-P32 required denaturing conditions. tr-P32 showed markedly higher expression and solubility than full-length P32. All recombinant proteins were immunoreactive with hyperimmune GTPV sera, as confirmed by western blotting. Native PAGE analysis of rA27L revealed concentration-dependent oligomerization into dimers, trimers and tetramers. Furthermore, in indirect ELISA, both A27L and P32 exhibited strong reactivity with sera from GTPV and SPPV infected or vaccinated animals at 1:50 dilution and did not show any cross-reactivity with related viruses. These results quantitatively demonstrated that baculovirus-expressed A27L and P32 proteins are superior diagnostic antigens, providing higher sensitivity and specificity for <em>Capripoxviruses</em> serodiagnosis and offering potential as improved candidates for both serological and prophylactic applications.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115234"},"PeriodicalIF":1.6,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144756864","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

Chimeric snakehead rhabdoviruses and zebrafish as a model system for investigating the role of fish rhabdoviral glycoproteins in in vivo virulence 以斑马鱼嵌合蛇头横纹鱼病毒糖蛋白为模型系统研究鱼类横纹鱼病毒体内毒力的作用

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-07-26 DOI: 10.1016/j.jviromet.2025.115232

Jae Young Kim, Mariem Bessaid, Kyung Min Lee, Ki Hong Kim

{"title":"Chimeric snakehead rhabdoviruses and zebrafish as a model system for investigating the role of fish rhabdoviral glycoproteins in in vivo virulence","authors":"Jae Young Kim, Mariem Bessaid, Kyung Min Lee, Ki Hong Kim","doi":"10.1016/j.jviromet.2025.115232","DOIUrl":"10.1016/j.jviromet.2025.115232","url":null,"abstract":"<div><div>Fish rhabdoviruses pose significant threats to aquaculture due to their pathogenicity and economic impact. This study explores the role of viral glycoprotein (G) and non-virion (NV) genes in determining virulence by generating a panel of chimeric snakehead rhabdoviruses (SHRVs) that express heterologous G or NV genes from pathogenic fish rhabdoviruses. Zebrafish (<em>Danio rerio</em>), which shows susceptibility to multiple rhabdoviruses and low mortality upon SHRV infection, was employed as an <em>in vivo</em> model to evaluate virulence. All chimeric SHRVs replicated efficiently in ZF-4 cells, indicating functional compatibility of the heterologous glycoproteins with the SHRV backbone. However, in the <em>in vivo</em> virulence analysis, G gene substitutions alone, even from highly virulent parental viruses (e.g., VHSV Ia, SVCV), did not significantly enhance mortality, suggesting that the G protein is insufficient by itself to confer high virulence in zebrafish. In contrast, a chimeric SHRV harboring the NV gene from VHSV Ia demonstrated elevated mortality and viral replication <em>in vivo</em> compared to wild-type SHRV, indicating the NV gene's role as a virulence determinant. These findings underscore the complexity of rhabdoviral virulence mechanisms and support the utility of the zebrafish–SHRV system for dissecting gene-specific contributions to pathogenicity in aquatic hosts.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115232"},"PeriodicalIF":1.6,"publicationDate":"2025-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144723737","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

Beyond the limits of conventional ‘endpoint’ ELISA and rescuing the signal with lag k-ELISA 超越了传统的“终点”酶联免疫吸附试验的限制，用滞后k-酶联免疫吸附试验挽救了信号

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-25 DOI: 10.1016/j.jviromet.2025.115231

Ksenia Katsanovskaja , Federica Marchesin , Jacquie Ujetz , Samreen Ijaz , Richard Tedder , Myra McClure

{"title":"Beyond the limits of conventional ‘endpoint’ ELISA and rescuing the signal with lag k-ELISA","authors":"Ksenia Katsanovskaja , Federica Marchesin , Jacquie Ujetz , Samreen Ijaz , Richard Tedder , Myra McClure","doi":"10.1016/j.jviromet.2025.115231","DOIUrl":"10.1016/j.jviromet.2025.115231","url":null,"abstract":"<div><div>Conventional colorimetric e-ELISA assays are widely used for quantifying antibodies in serum but are often labour-intensive, costly, and require serial dilutions for high-titre samples to ensure accuracy. To address these limitations, we propose the lag k-ELISA, a novel method for assessing antibody concentrations against SARS-CoV-2 in blood samples. This technique simplifies workflows while reducing time and labor compared to traditional ELISA. In this study, lag k-ELISA was applied to 169 sera for anti-S1 IgG and 79 samples for anti-S1 IgM assessment. Distinct analytical workflows were developed to process kinetic signal data, allowing for the description of Ab content. Key signal metrics—including time at OD minima, k-rates, and signal kinetics—proved effective in distinguishing samples by dilution factor or unitage. The assay’s performance was systematically evaluated under different operating conditions, such as stirring, readout time, and temperature, which significantly impacted assay outcomes and needed to be optimised to enhance the resolving power of lag k-ELISA. Our findings demonstrate that lag k-ELISA is a robust, efficient complement to traditional ELISA methods, offering reliable results with reduced labour, time, and costs. This technique is particularly advantageous for high-titre samples, streamlining antibody quantification workflows without compromising accuracy.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115231"},"PeriodicalIF":2.2,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144711023","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

Targeted enrichment of Elephant Endotheliotropic Herpesvirus for complete genome sequencing in elephants 大象嗜内皮疱疹病毒的靶向富集用于大象全基因组测序

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-24 DOI: 10.1016/j.jviromet.2025.115230

Kirtika Sharma , K.G. Sai Balaji , Gaurav Kumar Sharma , Abhijit M. Pawde , Sonalika Mahajan , Ravikant Agrawal , Pracheta Janmeda , Karikalan Mathesh

{"title":"Targeted enrichment of Elephant Endotheliotropic Herpesvirus for complete genome sequencing in elephants","authors":"Kirtika Sharma , K.G. Sai Balaji , Gaurav Kumar Sharma , Abhijit M. Pawde , Sonalika Mahajan , Ravikant Agrawal , Pracheta Janmeda , Karikalan Mathesh","doi":"10.1016/j.jviromet.2025.115230","DOIUrl":"10.1016/j.jviromet.2025.115230","url":null,"abstract":"<div><div>Elephant Endotheliotropic Herpesvirus (EEHV) poses a critical threat to young Asian (<em>Elephas maximus</em>) and African elephants (<em>Loxodonta africana</em>), with high mortality rates due to acute haemorrhagic symptoms from vascular endothelial damage. EEHV remains dormant in adult elephants, reactivating under stress or immune suppression. There are multiple genotypes of EEHV have been reported based on genetic heterogeneity. The diversity among EEHV subtypes, such as EEHV1A and EEHV1B, influences disease severity and host susceptibility. EEHV genomes are large and encode over 115 open reading frames, contributing to viral replication and immune evasion. Hence, generation of complete genome is essential to determine the genotypes of EEHV. However, the inability to culture EEHV <em>in</em>-<em>vitro</em> limits the study of its life cycle and pathogenesis, necessitating molecular techniques for direct analysis. Additionally, variable viral load in the infected tissues limits the success of DNA sequencing using direct sequencing. Hence, in this study several methods of viral or DNA enrichment were considered, and ultracentrifugation method was adopted to concentrate viral copies for improved genomic analysis. Ultracentrifugation demonstrates effectiveness in isolating EEHV, significantly enriching viral DNA for detection and characterization. This technique facilitates advancements in diagnostics, therapeutic development, and vaccine research, addressing the challenges of low viral loads and host DNA contamination in clinical samples.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115230"},"PeriodicalIF":2.2,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144711024","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

Genomic and molecular epidemiological characterization of a novel piscine poxvirus in the red seabream, Pagrus major 红鲷一种新型鱼痘病毒的基因组和分子流行病学特征

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-23 DOI: 10.1016/j.jviromet.2025.115229

Naritoyo Ishibashi , Yuri Akase , Shuntaro Watanabe , Hiroshi Yokoyama , Tohru Mekata

{"title":"Genomic and molecular epidemiological characterization of a novel piscine poxvirus in the red seabream, Pagrus major","authors":"Naritoyo Ishibashi , Yuri Akase , Shuntaro Watanabe , Hiroshi Yokoyama , Tohru Mekata","doi":"10.1016/j.jviromet.2025.115229","DOIUrl":"10.1016/j.jviromet.2025.115229","url":null,"abstract":"<div><div>Since 2020, unexplained mortality has been recorded in a red seabream hatchery. Affected juveniles exhibit skin darkening and sleep-like symptoms, which occasionally cause mass mortality. In this study, we detected a novel poxvirus genome in diseased fish via next-generation sequencing. Transcriptome analysis of moribund individuals revealed multiple contigs with sequence homology to the salmon gill poxvirus, suggesting the presence of a related viral agent. Subsequent metagenomic analysis led to the assembly of a 308-kbp viral genome of a novel piscine poxvirus, designated Japanese seabream poxvirus (JSPV), containing 338 predicted open reading frames. DNA polymerase of JSPV shared 62 and 44 % amino acid identity with those of the salmon gill poxvirus and carp edema virus, respectively. Comparative genomic analysis of piscine poxviruses revealed that genome synteny was highly conserved in the central region. Subsequently, a PCR method was developed and <em>I7L</em>, which encodes the virion core cysteine protease, was successfully detected in all JSPV genogroups. Based on the partial sequence of <em>I7L</em>, JSPV was classified into two major genogroups: I and II. Genogroup I was further subdivided into genogroups Ia and Ib.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115229"},"PeriodicalIF":2.2,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144702814","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

Establishment and application of a SYBR Green-based absolute real-time quantitative PCR assay for chilli yellow ringspot virus 辣椒黄环斑病毒SYBR绿基绝对实时定量PCR方法的建立与应用

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-23 DOI: 10.1016/j.jviromet.2025.115227

Yu Li , Kuo Wu , Xia Du, Yongdui Chen, Jie Zhang, Zhongkai Zhang

{"title":"Establishment and application of a SYBR Green-based absolute real-time quantitative PCR assay for chilli yellow ringspot virus","authors":"Yu Li , Kuo Wu , Xia Du, Yongdui Chen, Jie Zhang, Zhongkai Zhang","doi":"10.1016/j.jviromet.2025.115227","DOIUrl":"10.1016/j.jviromet.2025.115227","url":null,"abstract":"<div><div>Chilli yellow ringspot virus (CYRSV), a new emerging pathogen, was first detected in Yunnan Province, China. This virus is a novel agent that causes severe disease in vegetable crops and has a tendency to spread widely. CYRSV infection manifests as yellowing, ringspot and necrosis of the leaves and fruits of tomato plants. Consequently, the development of a sensitive, rapid, and accurate detection system for virus identification and epidemiological surveillance is imperative. In this study, we developed a quantitative PCR (qPCR) assay using SYBR Green for the detection of CYRSV accumulation. To achieve sensitive and rapid detection of CYRSV in tomato, specific primers were designed based on the coding sequence of the CYRSV nucleocapsid protein (N) gene. These primers demonstrated excellent specificity, sensitivity, and reproducibility. After inoculation with CYRSV in a greenhouse, the amount of virus accumulated in tomato leaves was detected. A total of 62 tomato fruit samples were collected from Yuanmou city, Yunnan Province, China, between May and December 2024. Of these, 75.81 % (47/62) tested positive for CYRSV via this qPCR method. These results indicated that the established SYBR Green-based qPCR assay could accurately detect CYRSV in tomato leaves and fruits. In conclusion, our study provides a valuable diagnostic tool for CYRSV detection and epidemiological investigation.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115227"},"PeriodicalIF":2.2,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144711022","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

Detecting rubella virus-specific antibodies by cytopathic effect and immunoenzymatic microneutralization tests 细胞病变效应和免疫酶微中和试验检测风疹病毒特异性抗体

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-21 DOI: 10.1016/j.jviromet.2025.115228

Iris Medits-Weiss, Heidemarie Holzmann, Lukas Weseslindtner, Karin Stiasny

{"title":"Detecting rubella virus-specific antibodies by cytopathic effect and immunoenzymatic microneutralization tests","authors":"Iris Medits-Weiss, Heidemarie Holzmann, Lukas Weseslindtner, Karin Stiasny","doi":"10.1016/j.jviromet.2025.115228","DOIUrl":"10.1016/j.jviromet.2025.115228","url":null,"abstract":"<div><h3>Background</h3><div>Rubella in early pregnancy can cause severe congenital malformations, but can be prevented by vaccination. Immunity to rubella is usually assessed by the detection of rubella virus-specific antibodies. Neutralization tests (NTs) are of highest diagnostic significance, because they measure the effectiveness of antibodies to block viral infection.</div></div><div><h3>Objectives</h3><div>The ability of rubella virus to cause a cytopathic effect (CPE) is limited. Therefore, most NT formats are based on immunoenzymatic techniques detecting viral proteins in infected cells. As Vero cells have been described to be susceptible to rubella virus infection with a more pronounced CPE, we aimed to develop an NT with these cells.</div></div><div><h3>Methods</h3><div>We compared rubella NTs with different infectivity readout methods (immunoenzymatic vs. CPE) using 42 human polyclonal samples with a known IgG ELISA concentration (IU/ml).</div></div><div><h3>Results</h3><div>We found a strong positive correlation between rubella virus-specific ELISA IgG units and neutralization titers determined with the two different NTs as well as between the two NT formats.</div></div><div><h3>Conclusion</h3><div>NT titer determination using CPE as a readout for infectivity is a reliable and promising method for rubella serology.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115228"},"PeriodicalIF":2.2,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144696607","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