Journal of virological methods最新文献

{"title":"Corrigendum to “Generation of infectious clone of bovine adenovirus type I expressing a visible marker gene” [J. Virol. Methods 261 (2018) 139–146]","authors":"Jingjing Ren , Lu Zhang , Peng Cheng , Fan Zhang , Zehui Liu , Suresh K. Tikoo , Rui Chen , Enqi Du","doi":"10.1016/j.jviromet.2024.115088","DOIUrl":"10.1016/j.jviromet.2024.115088","url":null,"abstract":"","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115088"},"PeriodicalIF":2.2,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142864807","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":"Effect of time and temperature on the stability of HPV and cellular nucleic acid using simulated dry self-samples","authors":"Linzi Connor ,&nbsp;Anna Davey ,&nbsp;Janathan Danial ,&nbsp;Sharon Moncur ,&nbsp;Hana Elasifer ,&nbsp;Catriona Graham ,&nbsp;Kate Cuschieri","doi":"10.1016/j.jviromet.2024.115101","DOIUrl":"10.1016/j.jviromet.2024.115101","url":null,"abstract":"<div><h3>Background</h3><div>Self-sampling is now a key component within HPV-based cervical screening programmes to engage individuals and enhance participation. As self-sampling is relatively new, information on the influence of pre-analytical parameters such as transit-temperature and time between sampling and testing on HPV test results requires detailed investigation.</div></div><div><h3>Methods</h3><div>FLOQSwabs® and Evalyn Brushes® were used to assess HPV and cellular stability over a 30-week period (0w,4w,12w,30w) at 4 °C, ambient, and 37 °C.</div><div>Vaginal self-samples were simulated by inoculating the devices with an HPV16-positive cell-line suspension. Devices were tested using two DNA-based (Anyplex™ II HPV28, Papilloplex® HR-HPV), one mRNA-based (APTIMA HR-HPV,) and one in-house beta-globin qPCR assay.</div></div><div><h3>Results</h3><div>No loss of qualitative HPV detection was observed after 12-weeks storage at ambient or 4°C irrespective of device or assay. For DNA-based assays, no loss of qualitative HPV detection was observed over time (30w) irrespective of temperature/device. Loss of qualitative mRNA signal was observed when devices were stored at 37°C for 12-weeks or longer; however, no loss of detection was observed at 30-weeks when either device was stored at 4°C.</div></div><div><h3>Conclusion</h3><div>HPV nucleic acid is stable on proxies of self-taken samples, however, the duration of stability was affected by the device and storage conditions. Such differences should be considered when optimising self-sampling exercises.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115101"},"PeriodicalIF":2.2,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142864808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and implementation of a novel method for detecting hepatitis C virus resistance-associated substitutions to NS3, NS5A, and NS5B inhibitors in Linzhou, China","authors":"Cui Zhang ,&nbsp;Yugang Nie ,&nbsp;Jian Li ,&nbsp;Xiaoyu Ji ,&nbsp;Mengjie Han ,&nbsp;Rong Qin ,&nbsp;Yuqiu Liu ,&nbsp;Wenge Xing ,&nbsp;Maofeng Qiu ,&nbsp;Ning Li ,&nbsp;Zhongfu Liu","doi":"10.1016/j.jviromet.2024.115102","DOIUrl":"10.1016/j.jviromet.2024.115102","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis C virus (HCV) resistance-associated substitutions (RASs) have a significant impact on the treatment of HCV with direct-acting antivirals (DAAs). However, limited research has been conducted, and no standardized methods for detecting RASs in mainland China.</div></div><div><h3>Objectives</h3><div>To develop and apply a novel method for detecting HCV RASs in HCV RNA-positive patients in Linzhou, China.</div></div><div><h3>Study design</h3><div>In total, 103 HCV RNA-positive serum specimens and epidemiological questionnaires were collected. A PCR method for detecting HCV RASs encompassing the NS3 to NS5B region was developed.</div></div><div><h3>Results</h3><div>Demographic analysis revealed a predominance of females (66/103, 64.1 %), with an average age of 70 years. Genotype 1b (GT1b) (17/103, 16.5 %) and GT2a (86/103, 83.5 %) were identified. The prevalence of RASs was higher (17/17, 100 %) in GT1b than in GT2a (7/86, 8 %). In GT1b, a higher frequency of RASs was observed in the NS5B region (17/17, 100 %) than in the NS3 (14/17, 82 %) and NS5A (10/17, 59 %) regions. C316N was the most prevalent, followed by S122G (71 %) and R30Q (35 %).</div></div><div><h3>Conclusions</h3><div>We introduced an innovative approach for the detection of HCV RASs and provided a wealth of information on HCV RASs in Linzhou, China. The findings support the cautious selection of treatment regimens, potentially improving patient outcomes.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115102"},"PeriodicalIF":2.2,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142854072","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":"Corrigendum to “Development and validation of a VP7-specific EIA for determining the potency and stability of inactivated rotavirus vaccine” [J. Virol. Method 332 (2025) 115079]","authors":"Sung-Sil Moon ,&nbsp;Houping Wang ,&nbsp;Kimberly Brown ,&nbsp;Yuhuan Wang ,&nbsp;Theresa K. Bessey ,&nbsp;Harry B. Greenberg ,&nbsp;Baoming Jiang","doi":"10.1016/j.jviromet.2024.115100","DOIUrl":"10.1016/j.jviromet.2024.115100","url":null,"abstract":"","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115100"},"PeriodicalIF":2.2,"publicationDate":"2024-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142829179","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":"Expression and characterization of canine distemper virus hemagglutinin protein in suspension mammalian cells","authors":"Jiaxi Cai ,&nbsp;Zishu Li ,&nbsp;Yu Wang ,&nbsp;Shuren Fang ,&nbsp;Xiaohan Fang ,&nbsp;Xianghong Xue","doi":"10.1016/j.jviromet.2024.115098","DOIUrl":"10.1016/j.jviromet.2024.115098","url":null,"abstract":"<div><div>Hundreds of millions of the domestic dogs worldwide are routinely inoculated with the modified live vaccines for canine distemper virus (CDV) every year. However, the corresponding serological diagnostic and detections are always lacking, thus, there is an urgent demand to establish its unique diagnostic technologies to produce high-quality antigenic biomolecules. In the present study, the ectodomain (et) of CDV hemagglutinin (H) protein was firstly expressed in a soluble and secreted forms by an Expi293F transient transfection system based on its antigenic secondary structure analysis. The yields of CDV H(et) protein was 2.6 g/L with purity over 95 % in supernatant of Expi293F cells. The CDV H(et) protein elicited comparative antibodies levels to the CDV virions in rabbit by ELISA and neutralization test. The purified polyclonal antibodies of immunized with CDV H(et) protein recognized both wild-type and vaccine CDV strains. More importantly, the purified polyclonal antibodies of CDV H(et) protein revealed significantly higher viral neutralizing activity than those from CDV-3 virions, which highlighted that the critical role of CDV H protein to elicit viral specific and protective neutralizing antibodies. Taken together, the CDV H(et) protein produced in mammalian expression systems was high-quality and good immunogenicity, and would be with great potential to serve as a serological diagnostic antigen or a novel CDV subunit vaccine in future.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115098"},"PeriodicalIF":2.2,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813688","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":"Detection of SARS-CoV-2- specific antibodies in domestic cats using different ELISA tests","authors":"Keyla P. Lopez ,&nbsp;Konner R. Cool ,&nbsp;Dashzeveg Bold ,&nbsp;Natasha N. Gaudreault ,&nbsp;Bailey A. Roberts ,&nbsp;Emma Maag ,&nbsp;Juergen A. Richt ,&nbsp;Roman M. Pogranichniy","doi":"10.1016/j.jviromet.2024.115099","DOIUrl":"10.1016/j.jviromet.2024.115099","url":null,"abstract":"<div><div>The emergence of SARS-CoV-2 raised concerns about the potential for interspecies transmission, particularly among domestic animals. We evaluated the seroprevalence of SARS-CoV-2 antibodies in domestic cats from various sites in North America. A total of 216 serum samples collected between December 2019 and February 2022, were analyzed using four different enzyme-linked immunosorbent assays (ELISAs), including a commercial surrogate virus neutralization test (sVNT), a commercial double antigen test (dN ELISA), and two <em>in-house</em> developed indirect ELISAS based on receptor-binding domain (RBD iELISA) and the nucleocapsid (N iELISA) proteins, respectively. Seropositive samples in the commercial ELISAs were subject to virus neutralization test (cVNT) employing the Wuhan-like USA-WA1/2020 SARS-CoV-2 isolate. Our findings revealed that, 6 % (12/216) of the cat serum samples tested positive by the sVNT, while 4 % (9/216) tested positive for the dN-ELISA. Interestingly, the N iELISA showed a higher seroprevalence, with 31 % of the samples testing positive, possibly due to cross-reactive antibodies against the N protein of other coronavirus commonly found in cats. There was a high concordance between sVNT, cVNT, and RBD iELISA. Among positive sVNT cat serum samples, 75 % (9/12) exhibited neutralizing titers with all samples also being positive by RBD iELISA. Notably, the RBD iELISA and sVNT demonstrated high sensitivity and specificity in detecting SARS-CoV-2 antibodies (100 and 79 %; 100 and 90 %, respectively). In conclusion, our study provides important insights into the seroprevalence of SARS-CoV-2 antibodies in domestic cats, highlighting the potential for interspecies transmission and the need for continued monitoring of SARS-CoV-2 in animal populations.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115099"},"PeriodicalIF":2.2,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of an all-in-one rabies virus Bat-Clade genomic sequencing and host identification protocol","authors":"Fernanda Marques de Souza Godinho ,&nbsp;Aline Campos ,&nbsp;Rosana Huff ,&nbsp;Amanda Pellenz Ruivo ,&nbsp;Thales Bermann ,&nbsp;Milena Bauerman ,&nbsp;Franciellen Machado dos Santos ,&nbsp;Taina Machado Selayaran ,&nbsp;Artur Beineke Correa ,&nbsp;Raissa Nunes dos Santos ,&nbsp;Paulo Michel Roehe ,&nbsp;Gabriel da Luz Wallau ,&nbsp;Richard Steiner Salvato","doi":"10.1016/j.jviromet.2024.115097","DOIUrl":"10.1016/j.jviromet.2024.115097","url":null,"abstract":"<div><div>Rabies virus (RABV), remains a significant public health concern, with bat-maintained lineages accounting for all currently documented cases in Brazil. Despite the availability of pharmacological prophylaxis for humans and animals, the high genetic diversity of RABV in diverse natural bat hosts and continued circulation in multiple animals pose challenges for effective surveillance. Here, we developed and validated a novel, rapidly deployable amplicon-based sequencing approach for RABV genomic surveillance. This \"all-in-one\" protocol integrates whole RABV genome sequencing with host species identification through COI gene amplification and sequencing, addressing the challenges posed by RABV's high genetic diversity and complex transmission dynamics. We assessed the protocol's effectiveness by sequencing 25 near-complete RABV genomes from host species across four distinct families (Bovidae, Equidae, Felidae, and Microchiroptera) obtained from the Rabies Control and Surveillance Program from the Rio Grande do Sul State, Southern Brazil. The method achieved an average genome coverage of 91.4 % at a minimum 5x read depth, with a mean depth coverage of 816x across sequenced genomes. The results demonstrated significant Bat-Clade sublineage diversity, which was classified using the MADDOG RABV lineage system. The protocol successfully identified three bat species (<em>Tadarida brasiliensis</em>, <em>Desmodus rotundus</em>, and <em>Myotis nigricans</em>) among the samples, highlighting its capability for precise host identification. This study presents a powerful tool for high-resolution evaluation of RABV genomic features and host identification, enabling more targeted animal and human health interventions. This new approach has the potential to enhance RABV surveillance capabilities, contributing to more effective rabies control strategies within a One Health framework.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115097"},"PeriodicalIF":2.2,"publicationDate":"2024-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142801501","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":"Comparing methods to detect cellular proteins on the surface of HIV-1 virions","authors":"Deepa Chaphekar ,&nbsp;Claire Fernandes ,&nbsp;Arvin T. Persaud ,&nbsp;Christina Guzzo","doi":"10.1016/j.jviromet.2024.115096","DOIUrl":"10.1016/j.jviromet.2024.115096","url":null,"abstract":"<div><div>The surface of HIV-1 is embedded with numerous host-derived proteins. Characterizing these proteins can enhance knowledge of virus biology and potentially identify novel therapeutic targets. As many of these proteins are present in low abundance on virion surfaces, their identification can be hindered by inherent variables in the methods employed to detect them, including their varying assay sensitivities, sample processing, quantitative capacity, and experimental reproducibility. Here, we have compared the quantification of virion-incorporated proteins using conventional virus immunocapture assays and western blotting, alongside an emerging technique called flow virometry (FV). Using four different pseudovirus models that each express a human protein of interest (CD14, CD38, CD59 and CD162), we compared four experimental techniques for their ability to reliably quantify the incorporation of those four proteins onto virion surfaces. Our results shed light on the advantages and caveats of each technique for detecting virion-incorporated proteins and highlight the breadth in quantification for each technique under different experimental conditions. Protein detection with (FV) provided distinct advantages as it enabled highly reproducible quantifications, had the lowest sample requirements and reagent costs, and minimal hands-on experimental time. We additionally highlight some important considerations in experimental design when studying virion-incorporated proteins, such as the effect of different antibody clones, assay incubation times, and contributions of extracellular vesicles. Most importantly, our data illustrate the importance of using a combination of orthogonal approaches to detect virus-associated proteins, to enable reliable and reproducible quantification that accounts for individual assay biases.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115096"},"PeriodicalIF":2.2,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel linear B cell epitope of the porcine circovirus type 3 capsid protein identified by phage display technology","authors":"Shu-qing Yang ,&nbsp;Ke Yang ,&nbsp;Xin-ran Li ,&nbsp;Yi Zheng ,&nbsp;San-jie Cao ,&nbsp;Qi-gui Yan ,&nbsp;Xiao-bo Huang ,&nbsp;Yi-ping Wen ,&nbsp;Qin Zhao ,&nbsp;Sen-yan Du ,&nbsp;Yi- fei Lang ,&nbsp;Shan Zhao ,&nbsp;Chun Li ,&nbsp;Rui Wu","doi":"10.1016/j.jviromet.2024.115080","DOIUrl":"10.1016/j.jviromet.2024.115080","url":null,"abstract":"<div><div>Porcine circovirus type 3 (PCV3) is endemic in swine worldwide and causes reproductive disorders, dermatitis and nephrotic syndrome, and multi-organ inflammation. PCV3 capsid protein (Cap) can self-assemble into virus-like particles (VLPs), and is an ideal candidate for vaccines and diagnostic reagents.In this study, the recombinant PCV3 Cap protein was successfully expressed in <em>E. coli</em> by deleting the nuclear localization sequence (NLS). The PCV3 VLPs were observed by transmission electron microscopy, and its immunogenicity was evaluated in six-week-old female BALB/c mice. A monoclonal antibody was named mAb 2D6, and demonstrated strong reactivity and specificity to PCV3 Cap. The purified mAb 2D6 was further used for bio-panning to select phage expressing specific epitopes from phage-displayed 7 mer-peptide library. A novel linear B-cell epitope, recognized by mAb 2D6, was identified at the amino acid region 47–53 of Cap. The phage peptide sequences were analyzed using multiple sequence alignment and evaluated by peptide ELISA. These results provide insights for developing diagnostic tools and potential vaccines for PCV3.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115080"},"PeriodicalIF":2.2,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142786102","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":"Purification and characterization of kyasanur forest disease virus EDIII domain of major envelope glycoprotein","authors":"Manjima Das ,&nbsp;Vivek Kumar ,&nbsp;Rishav Madhukalya ,&nbsp;Rohit Gupta ,&nbsp;Vidushi Agarwal ,&nbsp;Shweta Choudhary ,&nbsp;Mandar Bhutkar ,&nbsp;Shailly Tomar ,&nbsp;Dilip Kumar ,&nbsp;Rajesh Kumar","doi":"10.1016/j.jviromet.2024.115089","DOIUrl":"10.1016/j.jviromet.2024.115089","url":null,"abstract":"<div><div>Current research efforts are underway to create novel approaches for the efficient diagnosis, monitoring, and mitigation of Kyasanur Forest Disease Virus (KFDV) infections. Flavivirus subunit-based vaccines based on envelope glycoprotein EDIII are now in preclinical and clinical research stages. Efficient purification and isolation methods for surface immunogenic viral antigens, including the recombinant envelope immunoglobulin-like domain III (rEDIII) protein, are crucial for the production and manufacturing of promising vaccine candidates that have been extensively assessed in previous literature. Here, we describe a method for high-yield expression, purification, and refolding of a KFDV rEDIII protein from a bacterial expression system. The KFDV rEDIII protein is extracted from the inclusion bodies in urea denaturing buffer followed by nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. The purified, denatured KFDV rEDIII protein was subsequently refolded using a step-wise gradient urea dilution via the dialysis method. The circular dichroism and Fourier transform infrared spectroscopy analysis confirms that the refolded KFDV rEDIII maintains the native secondary conformation majorly containing β-strands. Our study provides valuable insights into the design and expression strategies of rEDIII as a novel subunit vaccine candidate against KFDV.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"333 ","pages":"Article 115089"},"PeriodicalIF":2.2,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142786117","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}