PEDV、TGEV和PRoV多重RT-PCR检测方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2016.10.002

Abstract

Abstract:

In this study,a multiplex RT-PCR assay was established to differentially detect porcine epidemic diarrhea virus (PEDV),porcine transmissible gastroenteritis virus (TGEV) and porcine rotavirus (PRoV) after optimization of the reaction conditions.Three pairs of primers PEDV-N,TGEV-M and PRoV-VP6 were designed for specifically amplifying PEDV N gene,TGEV M gene and PRoV VP6 gene,respectively.The assay could specifically amplify PEDV,TGEV and PRoV,but not classical swine fever virus (CSFV),porcine foot and mouth disease virus (FMDV),pseudorabies virus (PRV),porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2).The detection limits of PEDV,TGEV and PRoV standard recombinant plasmids were 1.41×10³,1.41×10² and 1.41×10³ copies/μL,respectively.The repeated reaction under the same conditions obtained uniform results.The assay was used to detect a total number of 190 clinical samples,of which 42 (22.11%) samples were positive for PEDV,58 (30.53%) samples for TGEV and 34 (17.89%) samples for PRoV,and there were mixed infection among these viruses.The results indicated that this multiplex RT-PCR assay had the advantages of sensitivity,specificity and repeatability and provided a useful tool for differential detection and epidemiological investigation of PEDV,TGEV and PRoV.

Key words: porcine epidemic diarrhea virus; transmissible gastroenteritis virus; porcine rotavirus; multiplex RT-PCR; detection method

CLC Number:

LONG Fei-xiang, SHI Kai-chuang, ZHANG Zhen, YIN Yan-wen, CHEN Han-zhong, MO Sheng-lan. Establishment and Application of a Multiplex RT-PCR Assay for Detection of PEDV,TGEV and PRoV[J]. , 2016, 43(10): 2518-2526.

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Establishment and Application of a Multiplex RT-PCR Assay for Detection of PEDV,TGEV and PRoV

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