Development of Nano-qPCR assay for the diagnosis of Infectious Bovine Rhinotracheitis Virus

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2025-08-14 DOI:10.1016/j.jviromet.2025.115239

Lin Wang, Peili Hou, Hongmei Wang, Hongbin He

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引用次数: 0

Abstract

IBRV causes severe respiratory and reproductive diseases in cattle, leading to significant economic losses. To improve diagnostic efficiency, this study developed a Nano-qPCR assay targeting the conserved UL50 gene of IBRV. The assay demonstrated high specificity, with no cross-reactivity to other tested bovine pathogens. Sensitivity testing demonstrated that Nano-qPCR could detect IBRV at concentrations as low as 3.5 × 10⁰ copies/μL, whereas conventional qPCR failed to detect it at this level. The assay was validated using clinical samples (nasal/oral swabs, feces, blood, and tissues) from unvaccinated cattle suspected of IBRV infection. Among 68 samples, Nano-qPCR detected six additional positive cases missed by conventional qPCR, which were later confirmed as IBRV-positive by viral isolation in MDBK cells. This confirmed the superior sensitivity of Nano-qPCR. In conclusion, the Nano-qPCR assay offers a rapid, cost-effective, and highly sensitive method for IBRV detection, making it a promising tool for disease surveillance and control in the cattle industry.

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牛传染性鼻气管炎病毒纳米qpcr检测方法的建立。

IBRV在牛中引起严重的呼吸道和生殖疾病，造成重大的经济损失。为了提高诊断效率，本研究开发了一种针对IBRV保守UL50基因的纳米qpcr检测方法。结果表明，该方法具有高特异性，与其他牛病原体无交叉反应性。灵敏度测试表明，纳米qPCR可以在低至3.5 × 10⁰copies/μL的浓度下检测到IBRV，而传统qPCR在此水平下无法检测到。使用疑似IBRV感染的未接种疫苗牛的临床样本（鼻/口拭子、粪便、血液和组织）验证了该测定方法。在68份样本中，Nano-qPCR检测到6例常规qPCR遗漏的阳性病例，随后在MDBK细胞中通过病毒分离证实为ibrv阳性。这证实了纳米qpcr的高灵敏度。总之，纳米qpcr检测提供了一种快速、经济、高灵敏度的IBRV检测方法，使其成为养牛业疾病监测和控制的一种有前景的工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.