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用于小反刍兽疫病毒特异性检测的实时 RT-PCR 分析方法。

A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus.

机构信息

Institute for Animal Health, Ash Road, Pirbright, Woking, Surrey GU240NF, UK.

出版信息

J Virol Methods. 2011 Feb;171(2):401-4. doi: 10.1016/j.jviromet.2010.11.022. Epub 2010 Nov 30.

Abstract

Peste des petits ruminants virus (PPRV) causes a devastating disease of small ruminants present across much of Africa and Asia. Recent surveillance activities and phylogenetic analyses have suggested that the virus is an emerging problem as it is now being detected in areas previously free of the disease. As such, the virus not only is threatening small ruminant production and agricultural stability in the developing world, but also poses an economic threat to livestock in the European Union (EU) through introduction from European Turkey and North Africa. This report describes the development of a high throughput, rapid, real time RT-PCR method for the sensitive and specific detection of PPRV using robotic RNA extraction. This assay targets the nucleocapsid (N) gene of PPRV and has been shown to detect all four genetic lineages of PPRV in tissues, ocular and nasal swabs and blood samples collected in the field. The lowest detection limit achieved was approximately 10 genome copies/reaction, making this assay an ideal tool for the sensitive and rapid detection of PPRV in diagnostic laboratories.

摘要

小反刍兽疫病毒(PPRV)会导致小反刍动物的毁灭性疾病,目前在非洲和亚洲的大部分地区都有出现。最近的监测活动和系统发育分析表明,由于该病毒现在在以前无病的地区被检测到,因此它是一个正在出现的问题。因此,该病毒不仅对发展中国家的小反刍动物生产和农业稳定构成威胁,而且还通过从欧洲土耳其和北非的传入,对欧盟(EU)的牲畜构成经济威胁。本报告描述了一种使用机器人 RNA 提取的高通量、快速、实时 RT-PCR 方法,用于敏感和特异性检测 PPRV。该检测方法针对 PPRV 的核衣壳(N)基因,已被证明可检测组织、眼部和鼻腔拭子以及现场采集的血液样本中的所有四种遗传谱系的 PPRV。实现的最低检测限约为 10 个基因组拷贝/反应,这使得该检测方法成为诊断实验室中敏感和快速检测 PPRV 的理想工具。

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