Establishment and Application of Multiplex Two-step RT-PCR for Detection of Swine Main RNA Virus Pathogens

doi:10.7668/hbnxb.2010.06.008

Abstract

Abstract: To develop a multiplex RT-PCR test for detection of swine main RNA virus pathogens, three sets of primers were designed based on the conserved domains of E2 gene of CSFV, Nsp2 gene of PRRSV and E gene of JEV.The conditions of the PCR test were optimized, such as Mg²⁺and the concentration of the primers, and annea.1ing and extension step were merged into one step as well.The multiplex two—step PCR test amplified product with size of 576, 482 and 375 bp specific for PRRSV, CSFV and JEV respectively.To evaluate the multiplex RT—PCR assay, 50 clinical samples were detected.The data showed that the multiple PCR method being 93%coincidence withthe single PCRs for the presence of CSFV, PRRSV and JEV.The optimized multiplex RT-PCR is both sensitive, specific and rapid, providing a new method for the clinical diagnosis and epidemiological research of the three diseases.

Key words: lassical Swine fever virus, Porcine reproductive and respiratory syndrome virus, Japanese enceph.alitis virus：Multiplex two—step RT-PCR

CLC Number:

S858.28

LU Bin, WANG Yi-cheng, WU Run, YUAN Xiu-fang, XU Li-hua, LI Jun-xing, WANG Zhao-wen. Establishment and Application of Multiplex Two-step RT-PCR for Detection of Swine Main RNA Virus Pathogens[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(6): 38-43. doi: 10.7668/hbnxb.2010.06.008.

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http://www.hbnxb.net/EN/Y2010/V25/I6/38

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