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《Chinese Journal of Animal Health Inspection》 2007-08
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One-Step Real-Time Quantitative Reverse Transcriptase PCR (qRT-PCR) for the Detection of Peste des Petits Ruminants Virus

Jingyue Bao, Lin Li, Zhiliang Wang, Jinming Li, Chunju Liu (National Diagnostic Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao 266032, China)  
In this study, a rapid and specific TaqMan-based, one-step real-time quantitative reverse transcriptase PCR (qRT-PCR) is developed for the detection of peste des petits ruminants virus (PPRV). Primers and probe were designed based on the nucleocapsid protein gene sequence. The specificity of the assay was assessed by detecting PPRV isolates of different lineage. The analytical sensitivity of the real-time qRT-PCR assay was achieved through the construction of in-house PPRV cRNA standard for generation of a standard curve. The real-time qRT-PCR assay is capable to detect all PPR viruses and none of the other morbilliviruses. The sensitivity of the assay was found to be 810 RNA copies per reaction. And the assay has a linear range from 8.1×102 to 8.1×108 copies as well as excellent intra-assay and interassay reproducibilities.
【Fund】: 受农业部重大外来动物疫病诊断方法研究及防治技术推广储备项目;; 农业部948项目
【CateGory Index】: S855
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