Journal of virological methods最新文献_第5页

Corrigendum to “A high-sensitivity qPCR method for detecting residual vero cell DNA in rabies vaccine production” [J. Virol. Methods 338 (2025) 115217] 一种检测狂犬病疫苗生产中残余vero细胞DNA的高灵敏度qPCR方法的勘误[J]。性研究。方法338(2025)115217]。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-07-19 DOI: 10.1016/j.jviromet.2025.115226

MaríaP. Almario, Jorge Rivera, Catalina Páramo, Valentina Jaramillo, Yanira Chaparro Zulma Rocío Suárez-Moreno

引用次数: 0

A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid colorimetric detection of pepper mild mottle virus (PMMoV) 逆转录环介导等温扩增（RT-LAMP）快速比色检测辣椒轻度斑驳病毒（PMMoV）。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-17 DOI: 10.1016/j.jviromet.2025.115225

Anthony J. Gross , Salvador Lopez Jr. , Alexandra Rogers , Scott Adkins , Mya Breitbart

{"title":"A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid colorimetric detection of pepper mild mottle virus (PMMoV)","authors":"Anthony J. Gross , Salvador Lopez Jr. , Alexandra Rogers , Scott Adkins , Mya Breitbart","doi":"10.1016/j.jviromet.2025.115225","DOIUrl":"10.1016/j.jviromet.2025.115225","url":null,"abstract":"<div><div>Pepper mild mottle virus (<em>Tobamovirus capsici</em>, PMMoV) is a plant virus in the genus <em>Tobamovirus</em> that infects peppers and other members of the family <em>Solanaceae</em>. The virus is transmitted mechanically, poses a significant threat to crops globally, and is one of the most abundant viruses found in human feces and wastewater. Two colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect PMMoV, one targeting the RNA-dependent RNA-polymerase (PMMoV_RdRp) and the other targeting the coat protein (PMMoV_CP). Synthetic gBlock positive controls were used to determine the detection limit of each assay. PMMoV_RdRp detected PMMoV at concentrations greater than or equal to 100 copies/µL, the same sensitivity as the published RT-qPCR assay for this gene. In contrast, the detection limit of the PMMoV_CP RT-LAMP assay was an order of magnitude greater. Both assays were specific to PMMoV and did not amplify plant host tissue or related tobamoviruses. Since these RT-LAMP assays do not require specialized laboratory equipment and yield positive results within 20–30 min, they are advantageous for point-of-use testing. Overall, the RT-LAMP assays described here are sensitive, specific, and more rapid than existing methods for PMMoV detection and quantification and thus have potential widespread applications for agriculture, wastewater treatment assessment, recreational water quality testing, and food safety.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115225"},"PeriodicalIF":2.2,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667942","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

Monkeypox in the 21st century: Insights into its pathogenesis and public health implications 21世纪的猴痘：对其发病机制和公共卫生影响的见解

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-16 DOI: 10.1016/j.jviromet.2025.115223

Duong Thuy Linh , Muhammad Nadir Shabbir

{"title":"Monkeypox in the 21st century: Insights into its pathogenesis and public health implications","authors":"Duong Thuy Linh , Muhammad Nadir Shabbir","doi":"10.1016/j.jviromet.2025.115223","DOIUrl":"10.1016/j.jviromet.2025.115223","url":null,"abstract":"<div><div>Monkeypox, a zoonotic illness caused by the Monkeypox virus (MPXV), is a re-emerging disease that has resurfaced as a prominent global public health issue in the 21st century. The 2022 outbreak, previously limited to Central and West Africa, exhibited extraordinary global dissemination, prompting critical inquiries over the virus's transmission mechanisms and containment strategies. This paper analyzes monkeypox's pathophysiology, epidemiological trends, and public health ramifications, emphasizing diagnostic difficulties, underreporting, and the inequitable access to vaccines and therapies. The discussion encompasses the significance of genomic surveillance, One Health strategies, and the necessity for research on effective antivirals and vaccines. Substantial knowledge deficiencies, encompassing virus mutation hazards and a restricted comprehension of animal reservoirs, impede preventative initiatives. Mitigating future epidemics necessitates addressing these problems through improved diagnoses, equitable vaccination distribution, and enhanced surveillance. This paper examines the present status of Monkeypox research and delineates prospective directions to inform global public health policies.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115223"},"PeriodicalIF":2.2,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144656479","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

Retraction notice to “Monitoring of human cytomegalovirus glycoprotein B genotypes using real-time quantitative PCR in immunocompromised Chinese patients” [J. Virol. Methods, 160 (2009) 74–77] 《利用实时定量PCR技术监测中国免疫功能低下患者巨细胞病毒糖蛋白B基因型》的撤稿通知[J]。性研究。方法，160(2009)74-77]。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2025-07-15 DOI: 10.1016/j.jviromet.2025.115221

J. Fan, X. Zhang, X.M. Chen, H.N. Gao, M.F. Yang, H. Zhao, J.H. Hu, W.H. Ma

引用次数: 0

Establishment of an indirect ELISA for feline coronavirus antibody detection and serotype discrimination 建立猫冠状病毒抗体间接ELISA检测及血清型鉴别方法

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-13 DOI: 10.1016/j.jviromet.2025.115224

Xiaoman Lu , Yanxuan Hu , Xiaoyuan Chen , Yongyi Shen

{"title":"Establishment of an indirect ELISA for feline coronavirus antibody detection and serotype discrimination","authors":"Xiaoman Lu , Yanxuan Hu , Xiaoyuan Chen , Yongyi Shen","doi":"10.1016/j.jviromet.2025.115224","DOIUrl":"10.1016/j.jviromet.2025.115224","url":null,"abstract":"<div><div>Feline coronavirus (FCoV) is a highly contagious pathogen that is endemic to feline populations and is classified into two serotypes, I and II. Current diagnostic techniques are insufficient for distinguishing between these serotypes, which impedes effective surveillance and prevention efforts. In response to this limitation, we have developed an indirect enzyme-linked immunosorbent assay (ELISA) utilizing recombinant nucleocapsid (N) protein for the broad detection of FCoV, alongside receptor-binding domain (RBD) proteins specific to serotypes I (I-RBD) and II (II-RBD) for the purpose of serotype differentiation. The assay underwent systematic optimization, achieving high sensitivity (with detection limits of 1:64,000 dilution for N and I-RBD, and 1:32,000 for II-RBD) and specificity, exhibiting no cross-reactivity with feline calicivirus (FCV), feline herpesvirus (FHV), or feline parvovirus (FPV). The reproducibility of the assay was validated, with intra- and inter-assay coefficients of variation remaining below 10 %. Clinical validation conducted on 123 feline serum samples indicated a seroprevalence of 73.17 %, with the serotype distribution comprising 91.11 % serotype I, 1.11 % serotype II, 2.22 % mixed infections, and 5.56 % cases that could not be typed. This ELISA represents a rapid, cost-effective, and field-deployable method for large-scale FCoV surveillance and serotyping, thereby contributing to enhanced feline health management and epidemiological research.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115224"},"PeriodicalIF":2.2,"publicationDate":"2025-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144632225","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

Development and characterization of a mouse-panda chimeric antibody with neutralizing activity against canine distemper virus 具有犬瘟热病毒中和活性的小鼠-熊猫嵌合抗体的研制与鉴定。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-11 DOI: 10.1016/j.jviromet.2025.115222

Zhengguo Wang , Guishan Ye , Kuijing He , Cong Cai , Anding Zhang , Long Li , Li Han

引用次数: 0

A high-sensitivity qPCR method for detecting residual Vero cell DNA in rabies vaccine production 一种检测狂犬病疫苗生产中残留Vero细胞DNA的高灵敏度qPCR方法

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-10 DOI: 10.1016/j.jviromet.2025.115217

María P. Almario , Jorge Rivera , Catalina Páramo, Valentina Jaramillo, Yanira Chaparrro, Zulma Rocío Suárez-Moreno

{"title":"A high-sensitivity qPCR method for detecting residual Vero cell DNA in rabies vaccine production","authors":"María P. Almario , Jorge Rivera , Catalina Páramo, Valentina Jaramillo, Yanira Chaparrro, Zulma Rocío Suárez-Moreno","doi":"10.1016/j.jviromet.2025.115217","DOIUrl":"10.1016/j.jviromet.2025.115217","url":null,"abstract":"<div><div>Residual host cell DNA in biological products requires precise quantification to meet regulatory safety standards. This study presents the development and validation of a qPCR assay for detecting and quantifying residual Vero cell DNA in rabies vaccine formulations. A SYBR Green-based quantitative PCR (qPCR) assay targeting the Vero cell-specific alpha-satellite DNA sequence was developed and validated in accordance with regulatory guidelines. The method was evaluated for specificity, sensitivity, linearity, precision, and robustness using a certified genomic DNA standard. The assay exhibited high specificity with no amplification from non-target DNA. The method demonstrated a linear dynamic range from 0.064 to 1000 ng/mL (R² > 0.999) with an amplification efficiency of 96.3 %. Sensitivity analysis confirmed reliable detection well below the regulatory threshold of 10 ng per dose, with a calculated limit of quantification of 0.31 ng/mL. Intra- and inter-assay precision tests showed low variability (CV < 20 %), supporting assay reproducibility. Accuracy was confirmed through recovery experiments (93.33–117.33 %) and concordance with a reference method (kappa = 1). This validated qPCR method offers a sensitive, specific, and reproducible tool for monitoring residual Vero cell DNA in rabies vaccine production, ensuring regulatory compliance through robust quality control.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115217"},"PeriodicalIF":2.2,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144588234","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

Rapid and efficient generation of viral genome knock-in cell lines using the CRISPR-Cas9 system to produce infectious virus 利用CRISPR-Cas9系统快速高效地生成病毒基因组敲入细胞系以产生感染性病毒

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-08 DOI: 10.1016/j.jviromet.2025.115219

Pooja Bhatia , Aas Mohd , Ishika Agrawal , Harshita Katiyar , Amit Goel , Meghali Aich , Debojyoti Chakraborty , Naga Suresh Veerapu

{"title":"Rapid and efficient generation of viral genome knock-in cell lines using the CRISPR-Cas9 system to produce infectious virus","authors":"Pooja Bhatia , Aas Mohd , Ishika Agrawal , Harshita Katiyar , Amit Goel , Meghali Aich , Debojyoti Chakraborty , Naga Suresh Veerapu","doi":"10.1016/j.jviromet.2025.115219","DOIUrl":"10.1016/j.jviromet.2025.115219","url":null,"abstract":"<div><div>Several medically significant viruses are difficult to propagate with conventional laboratory host systems, limiting their availability for detailed characterization, antiviral screening, and functional studies. A range of methods can be used to generate viruses, such as creating sophisticated cell lines, organoid cultures, and the utilization of animal models. Here, we report the generation and characterization of CRISPR-Cas9 edited Huh7 stable cell lines engineered to carry and express overlength HBV genotypes A, B, C and D and full HEV genomes in the AAVS1 site. Viral polymerase inhibitors and IFN-α significantly reduced the production of viral genomes and proteins from the edited cells. The virus released by the edited cells was infectious in permissive cell lines and could be blocked by neutralizing antibodies. This approach can extend to other viruses, like HCV genotype 3, that are hard to culture or to culturable viruses, like Dengue, for vaccine production.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115219"},"PeriodicalIF":2.2,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144605866","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

Development and validation of oligonucleotide sets for real time RT-PCR detection and next generation sequencing, of re-emerged sylvatic Dengue virus 2 strains 重新出现的森林型登革病毒2株实时RT-PCR检测和下一代测序的寡核苷酸集的开发和验证。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-05 DOI: 10.1016/j.jviromet.2025.115218

Ndeye Aminata Dia , Mignane Ndiaye , Diamilatou Balde , Mouhamed Kane , Agathe Shella Efire , Gerald Mboowa , Fatou Thiam , Yahya Dieye , Moussa Dia , Gamou Fall , Ndongo Dia , Amadou Alpha Sall , Ousmane Faye , Oumar Faye , Moussa Moïse Diagne , Manfred Weidmann , Idrissa Dieng

{"title":"Development and validation of oligonucleotide sets for real time RT-PCR detection and next generation sequencing, of re-emerged sylvatic Dengue virus 2 strains","authors":"Ndeye Aminata Dia , Mignane Ndiaye , Diamilatou Balde , Mouhamed Kane , Agathe Shella Efire , Gerald Mboowa , Fatou Thiam , Yahya Dieye , Moussa Dia , Gamou Fall , Ndongo Dia , Amadou Alpha Sall , Ousmane Faye , Oumar Faye , Moussa Moïse Diagne , Manfred Weidmann , Idrissa Dieng","doi":"10.1016/j.jviromet.2025.115218","DOIUrl":"10.1016/j.jviromet.2025.115218","url":null,"abstract":"<div><div>Dengue virus (DENV) is one of the most prevalent arboviral threats worldwide. The virus is associated with a high health and economic burden mainly in tropical and subtropical regions. Available molecular tools however fail to correctly serotype and sequence sylvatic DENV-2 (DENV-2/GVI) which in known to circulate in forests in West Africa and Malaysia. The recent emergence of human case linked to this virus variant in Southern Senegal raises concerns about the correct detection and characterization of the virus for public health purposes. Here we developed and validate new sets of oligonucleotides for DENV-2 (DENV-2/GVI) detection, and next generation sequencing. Validations were carried out using epidemic DENV and sylvatic DENV-2 strains from the biobank of the WHO collaborating Center for Arboviruses and Haemorrhagic fevers. The presented approaches showed good performance to specifically detect sylvatic DENV-2 in both singleplex and multiplex PCR with other DENV serotypes respectively with a limit of detection of 68.85 and 133.21 RNA copies/reaction at 0.95 probability in a probit analysis. Additionally, tilling PCR primers were developed which yield a better genome coverage ranging from 93.9 % to 95.1 % for all processed DENV-2/GVI strains both on Illumina and Nanopore platforms and outperform previous schemes to efficiently amplified DENV-2/GVI strains. In summary the developed oligonucleotides will contribute to improving DENV surveillance and genomic epidemiology in endemic areas.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115218"},"PeriodicalIF":2.2,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144584273","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

Detection of cross-reactivity of antibodies to the N proteins of feline morbillivirus and canine distemper virus in Japanese cat plasma samples 日本猫血浆中猫麻疹病毒和犬瘟热病毒N蛋白抗体的交叉反应性检测

IF 2.2 4区医学

Journal of virological methods Pub Date : 2025-07-01 DOI: 10.1016/j.jviromet.2025.115216

Shwe Thiri Maung Maung Khin , Sheikhi Mohammad Jafar , Minglin Ju , Fumi Murakoshi , Tetsuya Furuya

{"title":"Detection of cross-reactivity of antibodies to the N proteins of feline morbillivirus and canine distemper virus in Japanese cat plasma samples","authors":"Shwe Thiri Maung Maung Khin , Sheikhi Mohammad Jafar , Minglin Ju , Fumi Murakoshi , Tetsuya Furuya","doi":"10.1016/j.jviromet.2025.115216","DOIUrl":"10.1016/j.jviromet.2025.115216","url":null,"abstract":"<div><div>Feline morbillivirus (FeMV) is a globally emerging virus that has been linked to chronic kidney disease (CKD) in infected cats. Immunological assays, such as enzyme-linked immunosorbent assay (ELISA), are important for studying the virus and monitoring its prevalence. A study using rabbit antiserum demonstrated antigenic cross-reactivity between nucleocapsid (N) proteins of FeMV and canine distemper virus (CDV), suggesting not only the risk of false-positive anti-FeMV antibody detection in ELISAs but also potentially false-positive FeMV antigen detection in Western blotting. To examine whether such cross-reactivity occurs in tests using cat plasma samples, we developed ELISAs using affinity-purified recombinant N proteins of FeMV and CDV with expression in <em>Escherichia coli</em> and tested 100 cat plasma samples collected from veterinary clinics in Japan. Twenty samples were found to be positive for anti-FeMV antibodies, while 6 were positive for anti-CDV antibodies. All these latter 6 samples were double-positive for anti-FeMV antibodies. Western blotting with the purified proteins confirmed the specificity of these antibodies to their target viral antigens. A reverse transcription-quantitative PCR assay with a detection limit of 100 copies failed to detect CDV genomic RNA in these 6 double-positive samples. These results strongly suggest the cross-reactivity between anti-FeMV N protein antibodies in cat plasma samples and the CDV N protein. This antigenic cross-reactivity should be considered in future studies using immunological methods employing FeMV or CDV N proteins, or antibodies targeting them.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"338 ","pages":"Article 115216"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144549253","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