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Establishment and Application of a Multiplex RT-PCR Assay for Differential Detection of Classical,Highly Pathogenic and Vaccine Strains of North American Genotype PRRSV
Authors:SHI Kai-chuang  XU Xin-ting  HU Jie  SU Yan-qiong  LU Wen-jun  CHEN Han-zhong
Institution:1. Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China; 2. College of Animal Science and Technology, Guangxi University, Nanning 530005, China
Abstract:To establish a rapid method for differential detection of classical, highly pathogenic and TJM-F92 vaccine strains of North American genotype porcine reproductive and respiratory syndrome virus (PRRSV), a multiple RT-PCR assay was established. In this assay, two pairs of primers were designed according to the genomic sequences of classical, highly pathogenic and TJM-F92 vaccine strains of PRRSV. The assay could only detect PRRSV, but not detect CSFV, FMDV, PRV and PCV2. The detection limit of the method was as little as 1.13×103 copies/μL of templates. The established assay was successfully used to detect 349 clinical samples and 119 samples were positive for PRRSV, of which 5 samples were positive for classical PRRSV (C-PRRSV), 107 samples for highly pathogenic PRRSV (HP-PRRSV) and 7 samples for TJM-F92 vaccine strain (V-PRRSV), while 7 samples were positive for HP-PRRSV and V-PRRSV. The results indicated that the established multiple RT-PCR assay could be used for differential detection and epidemiological investigation of PRRSV.
Keywords:porcine reproductive and respiratory syndrome virus (PRRSV)  North American genotype  multiplex RT-PCR  wild-type strain  vaccine strain  
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