中国畜牧兽医2016，Vol.43Issue(10)：2518-2526,9.DOI:10.16431/j.cnki.1671-7236.2016.10.002
PEDV、TGEV和PRoV多重RT-PCR检测方法的建立及应用
Establishment and Application of a Multiplex RT-PCR Assay for Detection of PEDV,TGEV and PRoV
摘要
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 VP 6 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),pseud-orabies virus (PRV),porcine parvovirus (PPV)and porcine circovirus type 2 (PCV2).The detec-tion limits of PEDV,TGEV and PRoV standard recombinant plasmids were 1.41×10 3 ,1.41×10 2 and 1.41 ×10 3 copies/μL,respectively.The repeated reaction under the same conditions obtained uniform results.The assay was used to detect a total number of 1 90 clinical samples,of which 42 (22.1 1%)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.关键词
猪流行性腹泻病毒/猪传染性胃肠炎病毒/猪轮状病毒/多重RT-PCR/检测方法Key words
porcine epidemic diarrhea virus/transmissible gastroenteritis virus/porcine rotavirus/multiplex RT-PCR/detection method分类
生物科学引用本文复制引用
龙飞翔,施开创,张珍,尹彦文,陈汉忠,莫胜兰..PEDV、TGEV和PRoV多重RT-PCR检测方法的建立及应用[J].中国畜牧兽医,2016,43(10):2518-2526,9.基金项目
广西水产畜牧科技项目(桂渔牧科201528017) （桂渔牧科201528017）
