Journal of virological methods最新文献_第10页

AI-driven strategies for enhancing Mpox surveillance and response in Africa 在非洲加强麻疹监测和应对的人工智能驱动战略

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-24 DOI: 10.1016/j.jviromet.2025.115270

David B. Olawade , Chiamaka Norah Ezeagu , Chibuike S. Alisi , Aanuoluwapo Clement David-Olawade , Deborah Motilayo Eniola , Temitope Akingbala , Ojima Z. Wada

{"title":"AI-driven strategies for enhancing Mpox surveillance and response in Africa","authors":"David B. Olawade , Chiamaka Norah Ezeagu , Chibuike S. Alisi , Aanuoluwapo Clement David-Olawade , Deborah Motilayo Eniola , Temitope Akingbala , Ojima Z. Wada","doi":"10.1016/j.jviromet.2025.115270","DOIUrl":"10.1016/j.jviromet.2025.115270","url":null,"abstract":"<div><div>Mpox, a zoonotic viral disease endemic to several African countries, has re-emerged as a significant public health concern, particularly in regions with limited healthcare resources. Current public health strategies in Africa fall short due to fragmented surveillance systems, delayed diagnostic capabilities, and inadequate resource distribution networks that cannot effectively respond to rapidly evolving outbreaks in remote and underserved areas. This narrative review explores the potential of Artificial Intelligence (AI) to enhance the management and control of Mpox in Africa. AI technologies, including machine learning and predictive analytics, can significantly improve early detection, surveillance, contact tracing, case management, public health communication, and resource allocation. AI-driven tools can analyze large datasets to identify outbreak patterns, automate contact tracing through mobile data, optimize treatment plans, and tailor public health messages to specific communities. However, the successful implementation of AI faces challenges, including limited digital infrastructure, data quality issues, ethical concerns, and the need for capacity building. Furthermore, ongoing research is essential to refine AI algorithms and develop culturally sensitive applications. This review emphasizes the need for investment in infrastructure, training, and ethical frameworks to fully integrate AI into public health systems in Africa. By addressing these challenges, AI can play a pivotal role in mitigating the impact of Mpox and enhancing the resilience of healthcare systems against future infectious disease outbreaks. This represents a novel comprehensive synthesis of AI applications specifically for African Mpox control, providing a critical framework for evidence-based implementation strategies in resource-limited settings.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115270"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155245","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 of a fluorescence isothermal recombinase polymerase amplification assay for rapid detection of feline calicivirus 荧光等温重组酶扩增法快速检测猫杯状病毒的建立。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-10-05 DOI: 10.1016/j.jviromet.2025.115275

Yujun Zhu , Shuzhou Huang , Bihong Huang , Yuexiao Lian , Tongyuan Zhang , Feng Cong , Miaoli Wu

{"title":"Development of a fluorescence isothermal recombinase polymerase amplification assay for rapid detection of feline calicivirus","authors":"Yujun Zhu , Shuzhou Huang , Bihong Huang , Yuexiao Lian , Tongyuan Zhang , Feng Cong , Miaoli Wu","doi":"10.1016/j.jviromet.2025.115275","DOIUrl":"10.1016/j.jviromet.2025.115275","url":null,"abstract":"<div><div>Feline calicivirus (FCV) is responsible for a highly contagious disease in domestic cats. FCV may cause multiple symptoms and even death to the infected cats. A simple and cost-effective real-time RPA assay was developed for rapid detection of FCV in clinical samples. In this study, specific primers and probe were designed from the genome of FCV that prevalent in south China. The real-time RPA assay was carried out at 39℃ for 20 min before signal analysis by the fluorescence detector. The specificity and sensitivity were thoroughly validated and the results showed that no cross-reaction with irrelevant pathogens were found during the amplification, indicating the good specificity of the new developed real-time RPA assay. RNA standards were constructed and diluted to evaluate the limit of detection. The results showed that the detection limit of the real-time RPA assay could achieve 100 copies/μl, suggesting the high sensitivity of the assay. Additionally, the real-time RPA assay showed excellent performance in clinical sample detection, when compared with a TaqMan qPCR assay. The detection rate of FCV was 38.5 % (57/148) for real-time RPA assay and it was a little higher than 37.2 % (55/148) of the qPCR assay. Taking all together, the real-time RPA assay had potential application of FCV detection in clinical diagnosis. In conclusion, the new developed real-time RPA assay has provided an alternative strategy for rapid and sensitive detection of FCV in laboratories and animal clinics, especially those with limited facilities.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115275"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145244609","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

Evaluation of two IgG-scFv bispecific antibodies for neutralizing Omicron variants of SARS-CoV-2 两种IgG-scFv双特异性抗体中和SARS-CoV-2 Omicron变体的评价

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-05 DOI: 10.1016/j.jviromet.2025.115258

Diana Hinojosa-Trujillo , Freddy Dehesa-Canseco , Melissa García-Vega , Verónica Mata-Haro , Mario Solís-Hernández , Mónica Reséndiz-Sandoval , Fanglei Zuo , Harold Marcotte , Jesús Hernández

{"title":"Evaluation of two IgG-scFv bispecific antibodies for neutralizing Omicron variants of SARS-CoV-2","authors":"Diana Hinojosa-Trujillo , Freddy Dehesa-Canseco , Melissa García-Vega , Verónica Mata-Haro , Mario Solís-Hernández , Mónica Reséndiz-Sandoval , Fanglei Zuo , Harold Marcotte , Jesús Hernández","doi":"10.1016/j.jviromet.2025.115258","DOIUrl":"10.1016/j.jviromet.2025.115258","url":null,"abstract":"<div><div>Bispecific antibodies (bsAbs) offer an alternative to monoclonal antibody (mAb) cocktails for addressing the loss of efficacy due to the rapid emergence of SARS-CoV-2 mutants. The structure and specificity of the parental antibodies influence the development of a highly neutralizing bsAb. To design an effective bsAb, the recognition of 44 single-chain fragment variable (scFv) antibodies against variants of SARS-CoV-2 was evaluated, along with an assessment of their ability to competitively bind to the receptor-binding domain (RBD) compared to the most potent neutralizing mAbs. From this analysis, three antibodies − 19n01, 01n21, and 01n27 − were identified for their broad recognition and non-competitive binding behavior. These antibodies were selected as the parental antibodies for the design of two bsAb. The bsAb bis L and bis H were engineered as IgG-scFv constructs, each with the secondary domain oriented differently to introduce new specificities. Both bsAbs retained the neutralizing capabilities of their parental antibodies in live-virus assays, neutralizing the Omicron variants BQ.1.1 and XBB.1.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115258"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145015731","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

Evaluating the effectiveness of a novel environmental decontamination system utilizing low-energy hyper-charged photoelectrons against coronavirus 评估一种利用低能量超电荷光电子对抗冠状病毒的新型环境净化系统的有效性。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-20 DOI: 10.1016/j.jviromet.2025.115269

Madeeha Afzal , Mark D.P. Willcox , Stephan Praet , Murray Mcdonald , Muhammad Yasir

{"title":"Evaluating the effectiveness of a novel environmental decontamination system utilizing low-energy hyper-charged photoelectrons against coronavirus","authors":"Madeeha Afzal , Mark D.P. Willcox , Stephan Praet , Murray Mcdonald , Muhammad Yasir","doi":"10.1016/j.jviromet.2025.115269","DOIUrl":"10.1016/j.jviromet.2025.115269","url":null,"abstract":"<div><div>The COVID-19 pandemic had profound economic and social effects across the globe. The present study evaluated the virus attenuation efficacy of an environmental decontamination system named photon-mediated electron emitter (PMEE) on aerosolized and surface-associated coronavirus. The intensity of hyper-charged photoelectrons emitted by the PMEE were measured over distances of 1–5 m using a photon-detection mapping device. The antiviral efficacy of the PMEE was tested against mouse hepatitis virus (MHV-1) ATCC/VR261. For aerosolised studies, the MHV-1 was aerosolized using an electronic diffuser in an enclosed booth. Virus particles were exposed to PMEE for 10 and 15 min. For surface studies, viruses were dried on steel and laminate surfaces and then exposed to the PMEE from distances of 1 and 5-meters. The antiviral potential of the PMEE was evaluated by culturing MHV-1 in A9 ATCC/CCL 1.4 cells using a plaque assay. PMEE emission strength ranged from 1.44 to 1.86 V inside the booth and 0.83–1.86 V outside. The average size of the generated aerosol particles was 3.0 ± 0.3 µm. After 10 min exposure, the virucidal effects against particles of 2.1 µm, 1.1 µm, and 0.65 µm pore sizes were 74.5 ± 11.1 %, 79 ± 4.9 %, and 96 ± 1.4 % respectively. On surfaces, a 1-minute exposure at 1 m resulted in a 60 ± 0.5 % reduction on steel and 43 ± 2.7 % on laminate. The PMEE-based system effectively reduced the infectivity of MHV-1 both in aerosols and on surfaces, demonstrating strong potential for environmental decontamination applications.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115269"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124454","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

Cryopreservation of chicken and duck tracheal rings and precision-cut lung slices: A promising tool for the rapid characterization of avian influenza viruses 鸡鸭气管环和精确切割肺片的低温保存：一种有前途的禽流感病毒快速表征工具

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-02 DOI: 10.1016/j.jviromet.2025.115257

Alessandra Napolitan , Elisa Mazzacan , Niccolò Fonti , Sofia Tomasoni , Erica Melchiotti , Claudia Zanardello , Lucrezia Vianello , Sami Ramzi , Valentina Panzarin , Marika Crimaudo , Ranieri Verin , Francesco Bonfante , Eva Mazzetto

{"title":"Cryopreservation of chicken and duck tracheal rings and precision-cut lung slices: A promising tool for the rapid characterization of avian influenza viruses","authors":"Alessandra Napolitan , Elisa Mazzacan , Niccolò Fonti , Sofia Tomasoni , Erica Melchiotti , Claudia Zanardello , Lucrezia Vianello , Sami Ramzi , Valentina Panzarin , Marika Crimaudo , Ranieri Verin , Francesco Bonfante , Eva Mazzetto","doi":"10.1016/j.jviromet.2025.115257","DOIUrl":"10.1016/j.jviromet.2025.115257","url":null,"abstract":"<div><div>Since its emergence in 1996, highly pathogenic avian influenza (HPAI) viruses of the A/Goose/Guangdong/1/96 lineage have diversified into multiple clades, culminating in the 2020–2021 global panzootic caused by H5N1 viruses of the clade 2.3.4.4b. Further reassortment events have significantly diversified the phenotypes of these viruses, underscoring the need for continuous monitoring and strain characterization to better adjust control measures and mitigate the impact of the disease in wild birds and poultry. Standardized, ready-to-use <em>ex vivo</em> tissue platforms for rapid phenotyping of avian influenza viruses (AIVs) offer a valid alternative to <em>in vivo</em> models that are financially, ethically and logistically demanding. We optimized explant production and cryopreservation protocols for chicken and duck tracheal organ cultures (cTOCs and dTOCs) and precision-cut lung slices (cPCLS and dPCLS), assessing post-thaw viability, histological integrity, and susceptibility to AIV infection. Trehalose supplementation of cryopreservation solutions based on dimethyl sulfoxide (DMSO) and fetal bovine serum (FBS) significantly improved tissue viability. Although cryopreserved tissues were less viable than the fresh explants, viral replication was similar and only a modest reduction in susceptibility to infection was observed. Finally, we used duck and chicken TOCs to assess the ability of cryopreserved explants to discriminate viruses based on their divergent fitness and host preference. These findings underscore the potential of cryopreserved TOCs and PCLS as additional tools for the phenotypic characterisation of emerging AIVs.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115257"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144997278","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

Potential of recombinant avian adeno-associated virus as a viral vector for CRISPR/Cas9 delivery to avian cells 重组禽腺相关病毒作为CRISPR/Cas9载体的潜力

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-09 DOI: 10.1016/j.jviromet.2025.115263

Takumi Terada , Sodai Fujii , Nanako Yamanishi , Ryota Kajihara , Tenkai Watanabe , Ryo Ezaki , Hiroyuki Horiuchi , Mei Matsuzaki

{"title":"Potential of recombinant avian adeno-associated virus as a viral vector for CRISPR/Cas9 delivery to avian cells","authors":"Takumi Terada , Sodai Fujii , Nanako Yamanishi , Ryota Kajihara , Tenkai Watanabe , Ryo Ezaki , Hiroyuki Horiuchi , Mei Matsuzaki","doi":"10.1016/j.jviromet.2025.115263","DOIUrl":"10.1016/j.jviromet.2025.115263","url":null,"abstract":"<div><div>While genome editing has been established in chickens, where cultured primordial germ cell (PGC) systems are available, the implementation of genome editing remains a major challenge in many other birds due to the lack of robust PGC culture methods. Therefore, the development of reliable and efficient tools can significantly accelerate precision genome modification in avian species. Here, we evaluated the applicability of recombinant avian adeno-associated virus (rA3V) as a delivery vector for a CRISPR/Cas9 construct in avian cells using <em>Staphylococcus aureus</em>-derived Cas9 (SaCas9) and single-guide RNA (sgRNA). Infection with rA3V particles carrying an EGFP expression cassette (rA3V-EGFP) successfully induced EGFP expression in chicken fibroblasts (DF-1) cells, with approximately 80 % EGFP-positive cells at the maximum multiplicity of infection (MOI = 10,000). In plasmid-based transfection experiments, sgRNAs targeting the chicken <em>tyrosinase</em> locus and SaCas9 exhibited DNA cleavage activity in DF-1 cells. Furthermore, infection with rA3V particles encoding these CRISPR components successfully introduced indel mutations into the <em>tyrosinase</em> gene in DF-1 cells, with a calculated indel frequency of approximately 5.4 % at MOI = 40,000 without drug selection. Although EGFP expression was observed in quail fibrosarcoma cells, the percentage of EGFP-positive cells was much lower than that in DF-1 cells. In addition, <em>in vivo</em> infection with rA3V-EGFP of the chicken blastoderm failed to induce EGFP expression in germline cells, even at the highest applicable viral dose. In summary, rA3V can be used as a genome-editing vector in birds, although further investigation of its infectivity and tropism is necessary to expand its applicability to diverse avian species.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115263"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145040574","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

A dual-target real-time PCR for proactive detection of Mpox variants 主动检测m痘变异的双目标实时PCR。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-09-05 DOI: 10.1016/j.jviromet.2025.115260

Tracy D. Lee , Alan O’Dwyer , Michael Chan , Branco Cheung , Frankie Tsang , John R. Tyson , Kathleen L. Kolehmainen , Natalie A. Prystajecky , Agatha N. Jassem

{"title":"A dual-target real-time PCR for proactive detection of Mpox variants","authors":"Tracy D. Lee , Alan O’Dwyer , Michael Chan , Branco Cheung , Frankie Tsang , John R. Tyson , Kathleen L. Kolehmainen , Natalie A. Prystajecky , Agatha N. Jassem","doi":"10.1016/j.jviromet.2025.115260","DOIUrl":"10.1016/j.jviromet.2025.115260","url":null,"abstract":"<div><div>In 2022, cases of Monkeypox virus (MPXV) in California contained a mutation in the TNF receptor gene (GR2G) that rendered the virus undetectable using a widely adopted public health diagnostic qPCR assay. This underscored the need for a dual-target PCR approach and prompted validation of a second target by the BCCDC Public Health Laboratory. In addition to the GR2G target validated in the original qPCR assay (and duplexed with the endogenous target human β-globin (HBG)), GP113 (OPG128) was identified and validated using both clinical samples and MPXV DNA controls. Mutations in GR2G and GP113 (found in the emerging clade Ib) were also addressed along with the updated target. The new triplex assay (GR2G/GP113/HBG) had 100 % inclusivity and 100 % accuracy with all clinical samples tested and did not cross-react with herpes simplex virus-1 or −2, varicella zoster virus, or enterovirus. It showed < 5 % coefficient of variance between replicates and had a limit-of-detection of 10 copies/μL for GR2G and GP113. The use of two targets presents redundancy against further mutations in MPXV and is recommended for use with all viral qPCR assays moving forward.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115260"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145015721","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 analytical validation of a quantitative PCR assay for the detection of Magellanic penguin herpesvirus 1 麦哲伦企鹅疱疹病毒1型定量PCR检测方法的建立及分析验证

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-08-28 DOI: 10.1016/j.jviromet.2025.115255

Maris J. Daleo , Laura A. Adamovicz , Karisa N. Tang , Matthew C. Allender

{"title":"Development and analytical validation of a quantitative PCR assay for the detection of Magellanic penguin herpesvirus 1","authors":"Maris J. Daleo , Laura A. Adamovicz , Karisa N. Tang , Matthew C. Allender","doi":"10.1016/j.jviromet.2025.115255","DOIUrl":"10.1016/j.jviromet.2025.115255","url":null,"abstract":"<div><div>Herpesviruses are associated with disease in several aquatic bird species, including penguins. Magellanic penguin herpesvirus 1 (MagHV1) was initially detected in 58.3 % of oiled Magellanic penguins (<em>Spheniscus magellanicus</em>) in South America presenting with respiratory distress characterized by a combination of necrohemorrhagic tracheitis, fibrinous air sacculitis, pneumonia, and death. Additional exploration is needed to understand how herpesviruses affect penguin health; however, there is currently a lack of rapid, sensitive, and specific methods for detecting and quantifying herpesvirus infections in this taxon. To address this problem, we developed a real-time quantitative PCR (qPCR) assay for the detection of MagHV1 in penguins. Using a commercial program, TaqMan-MGB primer-probes targeting the DNA polymerase gene were designed <em>in silico</em>. Inter- and intra-assay variability, dynamic range, limit of detection, and analytical specificity were assessed to validate the assay per MIQE guidelines. The resulting assay was highly specific for MagHV1, failing to amplify fifteen closely related avian herpesviruses. It performed with high efficiency (slope =-3.336, R<sup>2</sup> = 0.999, efficiency 99.40 %) and low inter- and intra-assay variability (coefficient of variation <u><</u> 1.67 % at all dilutions). Reaction efficiency was not impacted by the presence of penguin DNA from known-negative tracheal swabs. This qPCR assay has a linear range of detection from 10<sup>7</sup> to 10<sup>1</sup> viral copies per reaction and provides a valuable tool in the surveillance and characterization of MagHV1 epidemiology in penguins. This assay can further be used to detect asymptomatic birds and as an effective tool to monitor infectious individuals.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115255"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144933144","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

Evaluation of the Applied Biosystems™ TaqPath™ Seq HIV-1 Genotyping Kit for HIV-1 drug resistance testing from dried blood spot specimens 应用Biosystems™TaqPath™Seq HIV-1基因分型试剂盒对干血斑点标本进行HIV-1耐药检测的评价

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-10-08 DOI: 10.1016/j.jviromet.2025.115277

Obiageli Okafor , Jane Cameron , Andrea Garcia , Charmaine Hinahon , Carmen Salvador-Palomeque , Mitchell Starr , Philip H. Cunningham

{"title":"Evaluation of the Applied Biosystems™ TaqPath™ Seq HIV-1 Genotyping Kit for HIV-1 drug resistance testing from dried blood spot specimens","authors":"Obiageli Okafor , Jane Cameron , Andrea Garcia , Charmaine Hinahon , Carmen Salvador-Palomeque , Mitchell Starr , Philip H. Cunningham","doi":"10.1016/j.jviromet.2025.115277","DOIUrl":"10.1016/j.jviromet.2025.115277","url":null,"abstract":"<div><h3>Objectives</h3><div>Monitoring HIV drug resistance is crucial for HIV treatment success. Logistical challenges restrict the use of liquid venous blood specimens for HIV drug resistance (HIVDR) testing, but dried blood spots (DBS) offer a more accessible alternative. This study evaluates the performance of TaqPath™ Seq HIV-1 Genotyping Kit (TaqPath kit) for detecting resistance-associated mutations in the protease (PR), reverse transcriptase (RT), and integrase (INI) regions of HIV-1 pol gene using DBS specimens.</div></div><div><h3>Methods</h3><div>A total of 79 DBS samples collected from ART-naive and ART-experienced individuals with viral loads ranging from 2830 to 9,095,161 copies/mL and HIV-1 subtypes A, B, C, F, and CRF01_AE were genotyped with TaqPath kit. Performance was assessed using the WHO HIVDR assay validation criteria.</div></div><div><h3>Results</h3><div>The assay met the amplification sensitivity criterion, achieving 95.7 % for PR/RT and 92.7 % for INI in high viral load samples. However, lower viral load samples between 2000 and 5000 copies/mL demonstrated reduced sensitivity of 50 % and 30 % for the PR/RT and INI regions respectively. Intra- and inter-assay agreement exceeded 99 % and mutation detection showed strong correlation with the reference assay, identifying over 95 % of reference mutations.</div></div><div><h3>Conclusions</h3><div>The TaqPath kit met the WHO acceptance criteria for HIVDR assay validation, except in the INI region at low viral loads, where amplification sensitivity was below the required threshold. Dried blood spot specimens are compatible with TaqPath kit for baseline and treatment-failure HIVDR genotyping, offering a reliable alternative to plasma in resource-constrained and remote settings.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115277"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145266663","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

Comparative effects of three SARS-CoV-2 inactivation methods on cytokine detection using LEGENDplex™ bead-based immunoassays 三种SARS-CoV-2灭活方法对LEGENDplex™免疫检测细胞因子的比较效果

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-01-01 Epub Date: 2025-08-24 DOI: 10.1016/j.jviromet.2025.115244

Yifan Chen , Ting Zhang , Jie Zhang , Xixuan Dong , Lixiang Xue , Zhongnan Yin

{"title":"Comparative effects of three SARS-CoV-2 inactivation methods on cytokine detection using LEGENDplex™ bead-based immunoassays","authors":"Yifan Chen , Ting Zhang , Jie Zhang , Xixuan Dong , Lixiang Xue , Zhongnan Yin","doi":"10.1016/j.jviromet.2025.115244","DOIUrl":"10.1016/j.jviromet.2025.115244","url":null,"abstract":"<div><div>In virology-related studies, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it is routine to inactivate body fluid samples carrying the virus to reduce the spread of the virus and guarantee the safety of biobankers and researchers. However, inactivation treatments may affect the molecular structure of proteins in biological samples, and it is necessary to select an inactivation method that has the least impact on the target molecule associated with protein detection techniques. Cytometric Bead Array (CBA), a novel and powerful technology, allows the simultaneous quantification of up to 10–30 different soluble proteins from one sample, with a particular focus on various cytokines and chemokines in human body fluids. But only a few studies have investigated the effect of inactivation methods on relevant assays. Therefore, this study aims to investigate various viral inactivation methods and evaluate their impact on LEGENDplex™ bead-based immunoassays. By detecting eight plasma samples and eight ascites samples, we assessed the impacts of heat denaturation, γ-irradiation, and paraformaldehyde (PFA) inactivation methods on certain protein components in plasma and ascites by LEGENDplex™ bead-based immunoassays. The results showed that heat treatment and γ-irradiation treatment had little effect on LEGENDplex™ bead-based immunoassays in both blood and ascites, while PFA treatment resulted in changes in multiple cytokines and chemokines. Among twenty-six cytokines and chemokines, perforin, I-TAC (CXCL11), Eotaxin (CCL11), and MIP-3α (CCL20) were more vulnerable to heat denaturation, γ-irradiation, and PFA treatments. To sum up, our findings provide evidence to inform the selection of optimal inactivation methods for reliable cytokine profiling in SARS-CoV-2-infected samples.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"339 ","pages":"Article 115244"},"PeriodicalIF":1.6,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144895297","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}

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