Journal of virological methods最新文献_第5页

Search and prove of efficient inhibitors against papain-like protease from SARS-CoV-2 SARS-CoV-2抗木瓜蛋白酶有效抑制剂的寻找和证明。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-19 DOI: 10.1016/j.jviromet.2025.115329

Wenfa Zhang , Xiao Yuan Chen , Sheng-Xiang Lin

{"title":"Search and prove of efficient inhibitors against papain-like protease from SARS-CoV-2","authors":"Wenfa Zhang , Xiao Yuan Chen , Sheng-Xiang Lin","doi":"10.1016/j.jviromet.2025.115329","DOIUrl":"10.1016/j.jviromet.2025.115329","url":null,"abstract":"<div><div>Papain-like protease (PLpro) is a key protease encoded by SARS-CoV-2, essential for viral polyprotein processing, replicase-transcriptase complex assembly, and interference with host immune responses. New COVID-19 cases continue to emerge, and WHO has identified coronaviruses with similar PLpro structures as potential pandemic threats. Using molecular modeling, docking, and protease activity assays, we identified four potent inhibitors of SARS-CoV-2 /PLpro with IC₅₀ values of 6.96–20.21 µM. Their binding affinities, determined by fluorescence titration, yielded dissociation constants (KD) of 4.18–13.06, supporting their inhibitory activity. In silico toxicity studies suggest that most inhibitors have LD₅₀ values comparable to GRL0617, a known PLpro inhibitor. Structural analysis revealed that all inhibitors interact with key residues Asp<sup>164</sup>, Pro<sup>248</sup>, Tyr<sup>264</sup>, Tyr<sup>268</sup>, and Gln<sup>269</sup> of PLpro, but differ in specific hydrogen bonding and hydrophobic interactions. Notably, compound P636–0664 exhibits the most extensive interactions, forming 2 hydrogen bonds and 13 hydrophobic contacts. These findings provide a structural basis for the optimization of inhibitors targeting the SARS-CoV-2 PLpro, contributing directly to anti-coronavirus drug discovery.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115329"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145804807","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

Targeting Yellow-Fever Virus: Development of a specific aptamer to NS1 protein 靶向黄热病病毒：NS1蛋白特异性适配体的研制

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-17 DOI: 10.1016/j.jviromet.2025.115330

Mariane Izabella Abreu de Melo, Alessandra Nunes Duarte Miranda, Antero Silva Ribeiro de Andrade

{"title":"Targeting Yellow-Fever Virus: Development of a specific aptamer to NS1 protein","authors":"Mariane Izabella Abreu de Melo, Alessandra Nunes Duarte Miranda, Antero Silva Ribeiro de Andrade","doi":"10.1016/j.jviromet.2025.115330","DOIUrl":"10.1016/j.jviromet.2025.115330","url":null,"abstract":"<div><div>Yellow Fever Virus (YFV), a mosquito-borne flavivirus, remains a significant public health concern despite the availability of effective vaccines. Accurate differential diagnosis continues to be challenging due to the high antigenic similarity among flaviviruses such as dengue and Zika, which compromises the specificity of current serological assays. Aptamers have emerged as promising alternatives to antibodies for diagnostic applications because of their high specificity, thermal stability, and ease of synthesis. In this study, we developed a DNA aptamer (Flav5) targeting the nonstructural protein 1 (NS1) of YFV using capillary electrophoresis–based SELEX (CE-SELEX). After three selection rounds, aptamer pools were sequenced and analyzed. Specificity assays demonstrated minimal cross-reactivity of Flav5 with NS1 proteins from dengue virus (serotypes 1–4) and Zika virus. The aptamer exhibited a high binding affinity for YFV-NS1, with a dissociation constant (Kd) of 34.5 ± 8.2 nM, as determined by quantitative PCR. The three-dimensional structure of Flav5 was modeled and docked to the tetrameric YFV-NS1 using the HDOCK server, revealing a stable binding interface within the inter-subunit cavity of NS1, supported by high confidence scores (>0.95). The Flav5 aptamer demonstrates strong potential for incorporation into YFV-specific diagnostic platforms, particularly in regions where multiple clinically relevant flaviviruses co-circulate. Its combination of high affinity, specificity, and favorable molecular docking characteristics, position it as a promising candidate for point-of-care diagnostic tools.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115330"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145794314","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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Evaluation of dried blood spot testing for serological monitoring of epizootic and zoonotic pathogens in domestic pigs 家猪兽疫和人畜共患病原体血清学监测中干血斑点试验的评价。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-24 DOI: 10.1016/j.jviromet.2025.115336

Ranja Steinhauer , Eric Kübler , Stefan Gaugler , Cornel Fraefel , Julia Lechmann

{"title":"Evaluation of dried blood spot testing for serological monitoring of epizootic and zoonotic pathogens in domestic pigs","authors":"Ranja Steinhauer , Eric Kübler , Stefan Gaugler , Cornel Fraefel , Julia Lechmann","doi":"10.1016/j.jviromet.2025.115336","DOIUrl":"10.1016/j.jviromet.2025.115336","url":null,"abstract":"<div><div>Dried blood spots (DBS) constitute a stable, cost-efficient sampling matrix that can be collected in a minimally invasive manner. Although widely adopted in human medicine, their use in veterinary diagnostics remains limited. This study aimed to establish and validate DBS elution protocols for use in commercial ELISAs to detect antibodies against Hepatitis E virus (HEV), African swine fever virus (ASFV) and Aujeszky’s disease virus (ADV) in domestic pigs. DBS were prepared from EDTA blood, dried serum spots (DSS) from serum. Additional DBS samples were prepared after spiking blood from healthy pigs with antibodies. Various parameters were evaluated to establish the final elution protocols, i.e. number of disks, type and volume of elution buffer, incubation time of the disks in elution buffer, and volume of eluate used for detection. Once the final protocols were in place, for each pathogen 52 DBS were tested in three independent runs. The diagnostic performance was evaluated by comparing the ELISA results of DBS eluates with the corresponding serum or plasma samples. For HEV, only one out of 52 DBS samples qualitatively did not match the plasma result in any of the three runs. For ASFV, all 52 DBS samples matched the qualitative results of the corresponding liquid samples. For ADV, two samples yielded false negative results in all three runs. The results suggest that DBS represent a practical and reliable alternative to liquid blood samples for antibody detection in pigs. Further validation with field samples and large-scale testing is needed.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115336"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145834245","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

Corrigendum to “An alternative model for HEV infection in the HepaRG cell line” [J. Virol. Methods 340 (2026) 115307] “HepaRG细胞系中HEV感染的另一种模型”的更正[J]。性研究。方法340(2026)115307。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-24 DOI: 10.1016/j.jviromet.2025.115334

Stacy Gellenoncourt , Marie Pellerin , Aïlona Marcadet-Hauss , Roxanne Fouillé , Michel Rivoire , Guillaume Passot , Julie Lucifora , David Durantel , Nicole Pavio , Virginie Doceul

引用次数: 0

A rapid immunochromatographic assay for the detection of BK Polyomavirus in urine samples from hematopoietic stem cell transplant recipients 造血干细胞移植受者尿液样本中BK多瘤病毒的快速免疫层析检测。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-05 DOI: 10.1016/j.jviromet.2025.115321

Etienne Brochot , Etienne Paubelle , Ophélie Fourdinier , Baptiste Demey , Aurélien Aubry , Virginie Morel , Antoine Touzé , Sandrine Castelain , François Helle

{"title":"A rapid immunochromatographic assay for the detection of BK Polyomavirus in urine samples from hematopoietic stem cell transplant recipients","authors":"Etienne Brochot , Etienne Paubelle , Ophélie Fourdinier , Baptiste Demey , Aurélien Aubry , Virginie Morel , Antoine Touzé , Sandrine Castelain , François Helle","doi":"10.1016/j.jviromet.2025.115321","DOIUrl":"10.1016/j.jviromet.2025.115321","url":null,"abstract":"<div><div>Hemorrhagic cystitis (HC) following hematopoietic stem cell transplantation (HSCT) is a significant event that can lengthen hospital stays and the need for care. The causes of HC can be multiple, but BK Polyomavirus (BKPyV) is the main protagonist in frequency. Currently, the clinical tool widely used to detect the presence of the virus in urine is PCR-based viral genome testing. We have developed a new rapid test for the antigenic detection of BKPyV in urine. The aim of this diagnostic study was to retrospectively evaluate the performance of this new assay as compared to BKPyV PCR on urine from HSCT patients with suspected HC. 49 samples from 49 different patients were evaluated, 20 of whom presented with HC. Of these 20 samples, 19 (95 %) were positive by the rapid antigen test. Of the 37 BKPyV PCR-positive samples, 31 were above 7 log<sub>10</sub> copies/mL, including the 20 patients with HC. Thus, the overall performance of the rapid test for HC is greatly improved compared with PCR, with specificity rising from 62.1 % to 86.2 % and positive predictive value from 64.5 % to 82.6 %. In conclusion, this new tool could be implemented as a point-of-care test to rapidly confirm or rule out a suspicion of BKPyV-HC after HSCT.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115321"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145701370","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

Corrigendum to ‘optimization and proficiency testing of a pseudovirus-based assay for detection of HIV-1 neutralizing antibody in China’[journal of virological methods 185 (2012) 267– 275] “中国基于假病毒的HIV-1中和抗体检测方法的优化和熟练测试”的勘误表[journal of病毒学方法185(2012)267- 275]。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-08 DOI: 10.1016/j.jviromet.2025.115319

Jianhui Nie , Wenbo Wang , Zhiheng Wen , Aijing Song , Kunxue Hong , Shan Lu , Ping Zhong , Jianqing Xu , Wei Kong , Jingyun Li , Hong Shang , Hong Ling , Li Ruan , Youchun Wang

引用次数: 0

Development of a colloidal gold-based immunochromatographic strip assay for rapid detection of Gyrovirus homsa 1 (GyH1) 胶体金免疫层析条带法快速检测回旋病毒1型（GyH1）

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-12-04 DOI: 10.1016/j.jviromet.2025.115320

Qi Yang , Ziqiang Cheng , Tianxing Yan , Guihua Wang

{"title":"Development of a colloidal gold-based immunochromatographic strip assay for rapid detection of Gyrovirus homsa 1 (GyH1)","authors":"Qi Yang , Ziqiang Cheng , Tianxing Yan , Guihua Wang","doi":"10.1016/j.jviromet.2025.115320","DOIUrl":"10.1016/j.jviromet.2025.115320","url":null,"abstract":"<div><div>Gyrovirus homsa 1 (GyH1) is an established causative agent of transmissible viral proventriculitis (TVP) in chickens. This study developed a colloidal gold immunochromatography assay (GICA) for GyH1 detection. Polyclonal and monoclonal antibodies against recombinant VP1 protein were generated. Colloidal gold nanoparticles were synthesized using the trisodium citrate reduction method, yielding uniform particle. Monoclonal antibodies were conjugated to colloidal gold particles, while polyclonal antibodies served as capture reagents immobilized on the membrane. Optimal labeling conditions were determined as pH 8.8 with a monoclonal concentration of 7.2 μg/mL. The assay demonstrated specific recognition of GyH1with a detection limit of 10<sup>3</sup> copies/μL. Batch-to-batch reproducibility was confirmed across all test strip. In summary, the GyH1 GICA exhibits high specificity, sensitivity, and reproducibility, providing an effective and low-cost tool for monitoring GyH1 early-stage infection status and technical support for field detection applications.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115320"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145693123","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

Design and development of a diagnostic method for HPV-16 virus with nanoball DNA based on vertical flow assay 基于垂直流式试验的纳米球DNA诊断HPV-16病毒方法的设计与开发。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-04-01 Epub Date: 2025-11-13 DOI: 10.1016/j.jviromet.2025.115300

Mansoreh Abdolhosseini , Mohammad H. Ghahremani , Moloud Absalan , Somayeh Jalilvand , Farshid Zandsalimi , Mohammad Panji , Mona Arianejad , Gholamreza Tavoosidana

{"title":"Design and development of a diagnostic method for HPV-16 virus with nanoball DNA based on vertical flow assay","authors":"Mansoreh Abdolhosseini , Mohammad H. Ghahremani , Moloud Absalan , Somayeh Jalilvand , Farshid Zandsalimi , Mohammad Panji , Mona Arianejad , Gholamreza Tavoosidana","doi":"10.1016/j.jviromet.2025.115300","DOIUrl":"10.1016/j.jviromet.2025.115300","url":null,"abstract":"<div><div>The Human papillomavirus is one of the deadliest causes of cancer in women. This study aimed to design and develop a diagnostic method based on Rolling Circle Amplification (RCA) and vertical Flow Assay (VFA) for detection of HPV16. DNA nanoball synthesis was optimized using Phi29 DNA polymerase and probes. Nitrocellulose membranes were saturated with saline-sodium citrate buffer (SSC), and different probe concentrations were tested for optimal color production and intensity measurement. The results were checked with Image J software. Membrane blocking and washing buffer formulations were optimized to minimize background noise and increase test accuracy. Real-time PCR was used as a gold standard, confirming 39 HPV16-positive clinical samples. The assay was validated with positive HPV16 and HSV samples, and the sensitivity and specificity of the VFA-RCA method were determined. The limit of detection (LOD) for HPV16 was 0.5 ng, equivalent to 472 DNA copies/ml, with a sensitivity of 94 % and a specificity of 88 %. The RCA-VFA method offers a reliable, sensitive, and specific diagnostic approach for the early detection of HPV16, demonstrating potential to identify cancer at an early stage.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115300"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145530776","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 split GFP approach to assay SARS-CoV-2 spike-dependent cell fusion 分裂GFP法检测SARS-CoV-2刺突依赖性细胞融合

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-02-01 Epub Date: 2025-11-01 DOI: 10.1016/j.jviromet.2025.115296

M. Jane Morwitzer, Ying Yi Zheng, Heather Friberg, Jeffrey R. Currier

{"title":"A split GFP approach to assay SARS-CoV-2 spike-dependent cell fusion","authors":"M. Jane Morwitzer, Ying Yi Zheng, Heather Friberg, Jeffrey R. Currier","doi":"10.1016/j.jviromet.2025.115296","DOIUrl":"10.1016/j.jviromet.2025.115296","url":null,"abstract":"<div><div>The SARS-CoV-2 spike (S) protein plays a central role in viral entry through receptor binding and membrane fusion, making it a key target for therapeutic interventions. While existing assays for studying spike-mediated fusion can be complex and lack real-time monitoring, we present a split GFP-based cell fusion assay that provides a straightforward and adaptable platform for evaluating fusion dynamics. This assay utilizes a split GFP system in non-adherent 293-F cells to detect fusion events, allowing for reliable assessment of spike-targeting monoclonal antibodies and fusion inhibitors. Our study demonstrates the effectiveness of this approach in evaluating the inhibitory potential of therapeutics against multiple SARS-CoV-2 variants. The results highlight the assay’s ability to distinguish variant-specific fusion characteristics and inhibitor responses, particularly in the context of Omicron’s altered entry pathways. By enabling the quantitative, real-time assessment of spike-mediated fusion, this platform provides a valuable tool for therapeutic screening, supporting future efforts to develop antiviral strategies against SARS-CoV-2 and other emerging coronaviruses.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"340 ","pages":"Article 115296"},"PeriodicalIF":1.6,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145431527","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

Optimization of rabies virus production in neuroblastoma cells using quality by design principles for vaccine and diagnostic applications 基于疫苗和诊断应用设计原则的神经母细胞瘤细胞狂犬病病毒生产质量优化

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-02-01 Epub Date: 2025-10-16 DOI: 10.1016/j.jviromet.2025.115289

Nayara Ugeda , Orlando Garcia Ribeiro , Elaine Raniero Fernandes , Fernanda Guedes , Sandriana Dos Ramos Silva , Iana Suly Santos Katz

{"title":"Optimization of rabies virus production in neuroblastoma cells using quality by design principles for vaccine and diagnostic applications","authors":"Nayara Ugeda , Orlando Garcia Ribeiro , Elaine Raniero Fernandes , Fernanda Guedes , Sandriana Dos Ramos Silva , Iana Suly Santos Katz","doi":"10.1016/j.jviromet.2025.115289","DOIUrl":"10.1016/j.jviromet.2025.115289","url":null,"abstract":"<div><div>Efficient production of high-titer rabies virus (RABV) stocks is critical for vaccine development, diagnostic assay design, and virological research. This study employed a Quality by Design (QbD) framework to optimize RABV propagation in Neuro-2a (N2a) cells, targeting infectious titers ≥ 8.0 log₁₀ FFU/mL and cell viability ≥ 85 %. Three genetically distinct RABV strains were evaluated: the laboratory-adapted CVS-11 strain for vaccine production and two field isolates, MN-18 and V-M, relevant to diagnostics and research. A 2 ³ full factorial Design of Experiments (DOE) was implemented to assess the impact of multiplicity of infection (MOI), incubation temperature, and culture medium composition on critical quality attributes (CQAs). Distinct strain-specific responses were observed: CVS-11 and V-M were significantly influenced by MOI (p < 0.05), whereas MN-18 demonstrated a more stable replication profile. Enriched medium significantly enhanced viral titers and cell viability across all strains (p < 0.001). Monte Carlo simulations (n = 10,000) delineated an optimal operational space—MOI 0.1–1.0, temperature 34–35°C, enriched medium—ensuring process robustness with 95 % confidence. This QbD-guided strategy provides a scalable, reproducible platform for generating high-quality RABV stocks, facilitating the manufacture of CVS-11-based vaccines and supporting the diagnostic use of field isolates as biologically standardized reference reagents for diagnostics.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"340 ","pages":"Article 115289"},"PeriodicalIF":1.6,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145318434","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