Validation of a molecular multiplex assay for the simultaneous detection and differentiation of bluetongue virus and epizootic haemorrhagic disease virus in biological samples

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-11-13 DOI:10.1016/j.jviromet.2024.115064

Ottavio Portanti, Eugenia Ciarrocchi, Roberta Irelli, Andrea Palombieri, Romolo Salini, Irene Melegari, Maura Pisciella, Simone Pulsoni, Daria Di Sabatino, Massimo Spedicato, Giovanni Savini, Alessio Lorusso

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Abstract

Bluetongue virus (BTV) and epizootic haemorrhagic disease virus (EHDV) are Culicoides-transmitted viruses, circulating in multiple serotypes, that cause two relevant WOAH-listed diseases of ruminants. Following its first identification in Tunisia in 2021, a novel EHDV strain belonging to serotype 8 has been detected in cattle showing BTV-like symptoms in Italy and Spain in 2022, and soon after in Portugal and France. These are European regions with recurrent circulations of different BTV serotypes. Hence, in this study we describe the validation of a TaqMan RT-qPCR panBTV/panEHDV assay, based on well-established primers and probes sets, able to simultaneously detect and distinguish between BTV and EHDV. The implemented assay, characterized by high sensitivity, specificity and good reproducibility, can be successfully applied for the rapid and affordable diagnosis needed in the current epidemiological situation and represents a powerful tool to be employed in surveillance and control strategies with a significant reduction of costs.

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验证同时检测和区分生物样本中蓝舌病病毒和流行性出血病病毒的分子多重分析法。

蓝舌病病毒（BTV）和附红细胞体出血病病毒（EHDV）是由蜱传播的病毒，以多种血清型流行，可引起两种被列入世界反刍动物卫生组织（WOAH）的相关疾病。继 2021 年在突尼斯首次发现后，2022 年又在意大利和西班牙安达卢西亚的牛群中检测到属于血清型 8 的新型 EHDV 株，这些牛群表现出类似 BTV 的症状，随后不久又在葡萄牙和法国检测到该病毒。这些欧洲地区反复出现不同血清型的 BTV。因此，在本研究中，我们介绍了基于成熟引物和探针组的 TaqMan RT-qPCR 泛 BTV/ 泛 EHDV 检测方法的验证情况，该方法能够同时检测和区分 BTV 和 EHDV。该检测方法灵敏度高、特异性强、重现性好，可成功应用于当前流行病学形势下所需的快速、经济的诊断，并可作为监测和控制策略的有力工具，显著降低成本。

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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.