Journal of virological methods最新文献

{"title":"Real time PCR-based evaluation of live attenuated lumpy skin disease virus vaccines for immunogenicity and efficacy","authors":"Mohamed Samy Abousenna,&nbsp;Heba A. Khafagy,&nbsp;Amal Abd El Moneim Mohamed,&nbsp;Sara El Sawy Ahmed,&nbsp;Fady Abd El Mohsen Shasha,&nbsp;Nermeen Gouda Shafik","doi":"10.1016/j.jviromet.2025.115327","DOIUrl":"10.1016/j.jviromet.2025.115327","url":null,"abstract":"<div><div>Lumpy skin disease (LSD) is a transboundary viral disease of cattle caused by lumpy skin disease virus (LSDV), resulting in substantial economic losses. Control of the disease relies primarily on live attenuated vaccines, making accurate potency assessment essential to ensure protective efficacy. Quantitative real-time PCR (qPCR) has emerged as a rapid and sensitive method for viral detection and quantification. In this study, ten commercial batches of live attenuated LSDV vaccines were evaluated using qPCR, and the molecular titers were compared with conventional tissue culture titration and in vivo protection in a calf challenge model. Tissue culture titration was performed on MDBK cells, qPCR quantified viral genome copy numbers, and in vivo challenge testing involved 56 calves. Statistical analyses, including Pearson and Spearman correlations and linear regression, were used to assess the relationships between qPCR-derived titers, tissue culture results, and protection outcomes. Eight of the ten vaccine batches met the WOAH-recommended potency range (3–4 log₁₀ TCID₅₀/mL), while two batches were suboptimal. qPCR-derived titers showed a strong correlation with tissue culture titers (Pearson’s r = −0.994; p &lt; 0.0001) r = −0.994; p &lt; 0.0001) and with protection outcomes following virulent challenge (r = 0.777; p = 0.008), with lower molecular titers corresponding to reduced protection. These results demonstrate that qPCR is a rapid, sensitive, and reliable surrogate for conventional potency testing, offering biosafe and time-efficient evaluation. Integrating molecular quantification into routine vaccine assessment can improve quality control, reduce reliance on in vivo challenge, facilitate harmonized vaccine deployment, and ultimately enhance field protection against LSD.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115327"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145781486","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 of sensitive and specific indirect ELISA tests for the detection of antibodies against bovine leukemia virus in serum and milk samples","authors":"Andrés Addiego ,&nbsp;Federico Carrión ,&nbsp;Natalia Olivero-Deibe ,&nbsp;Martín Fló ,&nbsp;Florencia Rammauro ,&nbsp;Natalia Ibañez ,&nbsp;Caroline da Silva Silveira ,&nbsp;Franklin Riet-Correa ,&nbsp;Lorena Tomé-Poderti ,&nbsp;Otto Pritsch ,&nbsp;Sergio Bianchi","doi":"10.1016/j.jviromet.2025.115335","DOIUrl":"10.1016/j.jviromet.2025.115335","url":null,"abstract":"<div><div>Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL), a disease characterized by lymphoproliferation and B-cell tumors. There is currently no available treatment or vaccine, making it a global animal health challenge and resulting in severe economic losses. The World Organization for Animal Health (WOAH) advocates for serological diagnostic tools and disease control programs to eradicate BLV. One major diagnostic technique recognized by the WOAH is the enzyme-linked immunosorbent assay (ELISA). We developed and optimized three indirect ELISAs for detecting anti-BLV antibodies in serum and milk. These assays utilize viral Env ectodomain and p24 recombinant proteins expressed in <em>Drosophila melanogaster</em> S2 cells and <em>Escherichia coli</em> systems, respectively. We compared their performance with a widely used commercial ELISA kit for completeness, which detects antibodies against BLV gp51 protein. Our immunoassay using recombinant p24 protein (anti-p24-S ELISA) showed comparable strength to the reference method (kappa index: 84.4 %, in 952 samples evaluated). Results for the anti-ectoEnv-S ELISA in serum samples closely matched the reference kit, with higher concordance than the anti-p24-S ELISA (kappa index: 94.9 %, in 1781 samples evaluated). Similarly, our in-house anti-ectoEnv-M ELISA in milk samples exhibited high concordance with both the commercial reference kit and the anti-ectoEnv-S (kappa index between the three tests: 95.6 %, in 479 blood and milk paired samples). Thus, we have developed, optimized, and validated three indirect in-house ELISAs for detecting anti-BLV antibodies in cattle, demonstrating good sensitivity and specificity values, using two different recombinant viral proteins.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115335"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145834283","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":"Evaluation of two commercial multiplex RT-PCR tests for broad genotypic detection of viral gastroenteritis","authors":"Chiara Filizzolo ,&nbsp;Mariangela Pizzo ,&nbsp;Marika Munacò ,&nbsp;Giovanni M. Giammanco ,&nbsp;Gianvito Lanave ,&nbsp;Vito Martella ,&nbsp;Floriana Bonura ,&nbsp;Simona De Grazia","doi":"10.1016/j.jviromet.2025.115328","DOIUrl":"10.1016/j.jviromet.2025.115328","url":null,"abstract":"<div><div>Acute gastroenteritis (AGE) remains a global health concern, particularly affecting children under five. While mortality rates have declined in industrialized countries, AGE continues to pose a significant burden on healthcare systems and remains a major threat in developing regions. This study evaluates the diagnostic performance of two commercial molecular assays for enteric viruses, GastroIntestinal (GI) Viral PLUS ELITe MGB® and GI Norovirus PLUS ELITe MGB®, for the detection of a broad panel of viral genotypes associated with AGE. A total of 222 archived stool samples, previously characterized by reference biomolecular methods, were analyzed. These included 50 rotavirus (RVA)-positive, 57 norovirus (NoV)-positive, 39 adenovirus (AdV)-positive, 45 astrovirus (HAstV)-positive samples, and 10 co-infections. Both assays showed excellent agreement with reference methods (κ ≥ 0.95), with sensitivity ranging from 99.4 % to 100 % and specificity from 98.1 % to 100 %, confirming their ability to detect a broad spectrum of circulating viral genotypes. Minor discrepancies were observed in NoV GII.6 strains, misclassified as Genogroup I (<em>GI</em>), likely due to overlapping annealing profiles. These findings support the use of GI Viral PLUS and GI Norovirus PLUS ELITe MGB® assays as reliable tools for rapid and accurate genotypic diagnosis of viral AGE. Their implementation may enhance patient management, reduce inappropriate antimicrobial use, and support timely outbreak containment, especially in healthcare settings.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115328"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145794348","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 of end-point and quantitative polymerase chain reaction assays to detect and discriminate between blueberry scorch virus and blueberry virus S","authors":"D.E.V. Villamor,&nbsp;D. Stainton,&nbsp;I.E. Tzanetakis","doi":"10.1016/j.jviromet.2025.115324","DOIUrl":"10.1016/j.jviromet.2025.115324","url":null,"abstract":"<div><div>Blueberry scorch virus (BlScV) and blueberry virus S (BVS) are closely related carlaviruses with the latter being only recently characterized. Most available PCR assays for BlScV fail to detect some isolates of the virus, as well as BVS. Here, we describe PCR assays designed based on all available virus sequences in public databases that can discriminate BlScV from BVS. Initially, a triplex endpoint PCR assay was developed that yields different amplicon sizes for each virus and incorporates an internal control which assesses the quality of the nucleic acids and the efficiency of the enzymatic reactions. Subsequently, a duplex quantitative PCR was designed employing primers targeting both viruses and discriminatory probes specific to each. Comparison between end-point and qPCR assays revealed near parallel sensitivity. The availability of the two PCR formats reliably improves the detection assays of the viruses in diagnostic facilities, quarantine, and clean plant programs.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115324"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145781285","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":"Evaluation of ∑-MM™ molecular medium for the SARS-CoV-2 RNA stability over 90 days under different temperature conditions","authors":"Giasemi C. Eptaminitaki,&nbsp;Alexandros Zafiropoulos,&nbsp;George Sourvinos","doi":"10.1016/j.jviromet.2025.115322","DOIUrl":"10.1016/j.jviromet.2025.115322","url":null,"abstract":"<div><div>The collection and storage of clinical specimens are critical determinants of reliable and accurate diagnostic outcomes. Swabs are commonly utilized for specimen collection aimed at the detection and identification of microorganisms. In this study, the ∑-MM™ Molecular Medium (Medical Wire and Equipment), which rapidly inactivates microorganisms, including bacteria, mycobacteria, and viruses, was evaluated for its capacity to preserve the stability of SARS-CoV-2 for up to 90 days under various temperature conditions. A total of 42 newly collected SARS-CoV-2 positive samples were selected to evaluate the performance of the ∑-MM™. Aliquots were stored under different conditions: at 4°C for 7 days, at room temperature for 7 days, at −80°C for one month, at room temperature for one month, at −80°C for three months, and at room temperature for three months. Samples were subsequently tested for SARS-CoV-2 RNA by Real-Time PCR. No statistically significant differences were observed across all storage conditions, when compared to the baseline measurements. The ∑-MM™ effectively preserves SARS-CoV-2 for up to 90 days at ambient temperatures, without statistically significant changes in CT values compared to baseline. These results validate ∑-MM™ as a reliable transport medium that enables specimen storage and transport under ambient conditions without compromising diagnostic accuracy. Given the demonstrated stability, future studies will explore the performance of ∑-MM™ over extended storage durations.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115322"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145701404","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":"Rapid establishment and validation of a PMAxx-RT-qPCR method for infectious titer detection of freeze-dried live attenuated hepatitis A vaccine (H2 Strain)","authors":"Yongrong Wang ,&nbsp;You Gao ,&nbsp;Wei Wang ,&nbsp;Qin Liu ,&nbsp;Yadong Li ,&nbsp;Haiyuan Zeng ,&nbsp;Lifei Chen ,&nbsp;Chen Cheng ,&nbsp;Fan Wu","doi":"10.1016/j.jviromet.2025.115333","DOIUrl":"10.1016/j.jviromet.2025.115333","url":null,"abstract":"<div><div>The infectious titer of hepatitis A virus (HAV) is a critical quality parameter for evaluating potency of freeze-dried live attenuated hepatitis A (HepA-<span>L</span>-fd) vaccines. In this study, we developed a rapid PMAxx-RT-qPCR for detecting the infectious titer of HepA-<span>L</span>-fd vaccines. This method enables virus titer detection to be completed within a day. Systematic optimization of experimental parameters yielded a validated methodology demonstrating excellent precision (coefficient of variation, CV &lt; 1 %), accuracy (recovery rate 107.6 ± 3.7 %), and a linear range of 6.23–7.73 log₁₀CCID₅₀/mL. Comparing with the measurements from the traditional method, results showed no significant differences among the six batches of vaccine finished products (P = 0.7452). This optimized PMAxx-RT-qPCR assay provides a rapid and reliable analytical means for the infectious titer assay in HepA-<span>L</span>-fd vaccines.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115333"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145834281","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":"Application of digital PCR and CRISPR/Cas13a-based fluorescent assay for accurate and on-site detection of cotton leafroll dwarf virus","authors":"Vijay Sheri ,&nbsp;Pankaj K. Verma ,&nbsp;SaiKrishna Lekkala,&nbsp;Madhusudhana R. Janga","doi":"10.1016/j.jviromet.2025.115332","DOIUrl":"10.1016/j.jviromet.2025.115332","url":null,"abstract":"<div><div>Cotton leafroll dwarf virus (CLRDV) is an emerging viral pathogen posing a significant threat to cotton production in the United States. Early and accurate detection is critical for effective disease surveillance and management. Although traditional reverse transcription PCR (RT-PCR) is commonly employed for CLRDV diagnosis, it suffers from limitations in sensitivity, quantification accuracy, and involves labor-intensive workflows. In this study, we evaluated two advanced molecular diagnostic approaches for detecting CLRDV, digital PCR (dPCR) and CRISPR/Cas13a-based fluorescent assay. Symptomatic cotton leaf samples from Lubbock and Brownfield, Texas, were screened and confirmed positive by RT-PCR. Digital PCR analysis enabled absolute quantification of viral load, revealing significantly higher titers in Brownfield (F2) samples and offered improved sensitivity over RT-PCR, particularly in samples with low viral loads. However, dPCR is resource-intensive and requires specialized instrumentation. To address the need for rapid, field-deployable diagnostics, we developed a CRISPR/Cas13a-based assay targeting the conserved ORF3, ORF2, and ORF3a regions of the CLRDV genome. Adapted from the SHERLOCK platform, this fluorescence-based assay uses collateral cleavage activity of Cas13a to enable highly specific visual detection. While the assay successfully enabled direct detection from crude leaf extracts without RNA purification, the sensitivity analysis was conducted using purified, <em>in vitro</em> transcribed RNA. Fluorescence signals were reliably observed with as few as 50 RNA copies, defining the assay’s practical limit of detection. While dPCR is optimal for quantitative laboratory analysis, the CRISPR/Cas13a-based assay offers a rapid, sensitive, and cost-effective tool for field-level detection. Together, these complementary tools enhance CLRDV surveillance and management in cotton.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115332"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145804761","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 of a rapid, real-time RT-RAA assay for sensitive detection of bovine enterovirus in fecal samples","authors":"Cuilan Wu ,&nbsp;Xixian Liu ,&nbsp;Lan Jia ,&nbsp;Zhijie Mo ,&nbsp;Yeheng Gao ,&nbsp;Shuhong Zhong ,&nbsp;Shiwen Feng ,&nbsp;Zhongwei Chen ,&nbsp;Xian Li ,&nbsp;Xindong Wang ,&nbsp;Xiongbiao Xuan ,&nbsp;Huili He ,&nbsp;Shuai Hu ,&nbsp;Changting Li ,&nbsp;Zuzhang Wei ,&nbsp;Yongping Xie ,&nbsp;Hao Peng ,&nbsp;Yingyi Wei ,&nbsp;Jun Li","doi":"10.1016/j.jviromet.2025.115331","DOIUrl":"10.1016/j.jviromet.2025.115331","url":null,"abstract":"<div><div>Bovine enterovirus (BEV) is a significant pathogen affecting bovine intestinal health, posing challenges to cattle production and public health. Existing detection methods, such as RT-PCR, RT-qPCR, and LAMP, are time-consuming and dependent on specialized laboratory facilities, highlighting the need for rapid and field-deployable diagnostic tools. In this study, we developed a real-time reverse transcription recombinase-aided amplification (RT-RAA) assay designed for BEV detection, capable of delivering results within 20 min at 39℃. The real-time RT-RAA assay showed no cross-reactivity to other eight pathogens, including BCoV, BVDV, BKV, BRV, BAstV, STEC, FMDV, and PEV. The sensitivity assay revealed that the real-time RT-RAA method could reliably detect a minimum plasmid concentration of 5.50 × 10 ¹ copies/reaction. Clinical validation conducted with 780 field samples revealed performance comparable to those of conventional RT-PCR and RT-qPCR methods, achieving a positivity rate of 23.59 %. The RT-RAA method operates at low temperatures, requires minimal instrumentation, and reduces technical demands, making it ideal for resource-limited settings. Its portability, cost-effectiveness, and robustness position it as a transformative tool for BEV surveillance, enhancing outbreak control, livestock health management, and food safety.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115331"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145804779","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":"Evaluation of Tasso T20 to collect dried blood for the quantification of HIV-1 RNA","authors":"Sara W.F. Geffert ,&nbsp;Soya S. Sam ,&nbsp;Karen Chandran ,&nbsp;Curt G. Beckwith ,&nbsp;Rami Kantor ,&nbsp;Angela M. Caliendo","doi":"10.1016/j.jviromet.2025.115326","DOIUrl":"10.1016/j.jviromet.2025.115326","url":null,"abstract":"<div><div>Tasso T20 (T20) is a finger stick-free, self-sampling device capable of at-home blood sample collection. We optimized HIV-1 RNA elution from T20, evaluated its analytical performance, compared its performance to Whatman dried blood spots (DBS), and assessed HIV-1 RNA stability. We used mock viral samples to optimize HIV-1 RNA elution using viral concentrations 10,000 and 3000 copies/mL. Limit of detection (LOD) and limit of quantification (LOQ) were determined, and linearity and reproducibility were verified. Analytical agreement between T20 and DBS was assessed using eight concentrations, from 100,000 to 1000 copies/mL. T20 HIV-1 RNA stability was assessed over time. All samples were tested using the m2000 RealTi<em>m</em>e HIV-1 DBS assay. The optimized elution conditions selected were 30 min of sonication followed by 30 min of incubation at 55 °C. The 95 % LOD and LOQ were determined to be 3.56 and 3.70 log₁₀ copies/mL, respectively. Good linearity was observed with a strong correlation (R²=0.98) and excellent reproducibility. Analytical agreement analysis demonstrated a mean difference of −0.36 log₁₀ copies/mL (SD ± 0.21 log₁₀ copies/mL). ANOVA indicated all samples remained stable (p value=0.33, F=1.22). The Tasso T20 is capable of quantifying HIV-1 viral loads, with results comparable to DBS.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115326"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145743229","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":"Alternatives to central nervous system samples for enhanced rabies surveillance: Exploring natural infections of Lyssavirus rabies in striped skunks (Mephitis mephitis) in Northern Colorado","authors":"Clara C.P. Mankowski ,&nbsp;Matthew W. Hopken ,&nbsp;Terry R. Spraker ,&nbsp;Amy T. Gilbert","doi":"10.1016/j.jviromet.2025.115325","DOIUrl":"10.1016/j.jviromet.2025.115325","url":null,"abstract":"<div><div>Enhanced rabies surveillance (ERS) of wildlife populations complements passive public health surveillance through active field-based sampling. The combined data augment rabies management decision-making through tracking virus distribution and movement, rapidly identifying potential outbreaks, and informing containment and elimination strategies at regional and landscape scales. Laboratory diagnostic methods are critical to ERS and several gold standard methods are recommended for detection of <em>Lyssavirus rabies</em> (RV); each validated using central nervous system (CNS) tissue which requires necropsy and cold storage, posing challenges and risks in remote field settings. Alternative sample types to CNS tissue may improve ERS efficiency and field personnel safety, provided they offer robust diagnostic specificity and sensitivity. We evaluated multiple sample types opportunistically collected postmortem from naturally infected striped skunks (<em>Mephitis mephitis</em>) and used real-time reverse transcriptase PCR (rtRT-PCR) to evaluate diagnostic sensitivity and specificity of each sample type compared to CNS tissue. The detection rate of RV RNA was 97 % (95 % CI 81–100) for oral swabs and 96 % (95 % CI 80–100) for eye swabs; when swab types were combined, the detection rate was 100 % (95 % CI 85–100) and comparable to detection from CNS. Other sample types demonstrated compromised diagnostic sensitivities (33–76 %). All sample types were 100 % specific for RV diagnosis. The combination of eye and oral swabs as ERS samples may expand sampling opportunities that improve efficiency, mitigate sample collection and storage challenges, and decrease RV exposure risk among field staff while enhancing the information provided to estimate impacts of RV management on target wildlife populations.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"341 ","pages":"Article 115325"},"PeriodicalIF":1.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145724124","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}