中国畜牧兽医2016，Vol.43Issue(9)：2455-2460,6.DOI:10.16431/j.cnki.1671-7236.2016.09.034
猪流行性腹泻病毒与猪伪狂犬病病毒二重 RT-PCR 检测方法的建立及应用
Establishment and Application of Duplex RT-PCR Assay for Detection of Porcine Epidemic Diarrhea Virus and Pseudorabies Virus
摘要
Abstract
In order to establish an assay for detecting porcine epidemic diarrhea virus (PEDV)and pseudorabies virus (PRV),two pairs of primers were designed basing on the M gene of PEDV and gE gene of PRV,respectively.The total RNA of standard PEDV and PRV strains were used as templates to establish the duplex RT-PCR assay.The specificity,sensitivity,repetition and clinic detection of the established assay were tested.The result revealed that the threshold of duplex RT-PCR was 10 TCID50/mL of PEDV and PRV,and no products were amplified from the cell or the nucleic acid of other 7 kinds of pathogenic viral or bacterial microorganism.The detection re-sults for 26 clinical suspicious PEDV or PRV infected pigs were consistent with the results tested by sequencing.This study suggested that the duplex RT-PCR method was highly specific,repeat-able and sensitive,and was suitable for clinic rapid differential diagnosis of PEDV and PRV.关键词
猪流行性腹泻病毒/猪伪狂犬病病毒/二重 RT-PCRKey words
porcine epidemic diarrhea virus/pseudorabies virus/duplex RT-PCR分类
农业科技引用本文复制引用
孙彦琴,马震原,闫若潜,王东方,曹伟伟,郭育培..猪流行性腹泻病毒与猪伪狂犬病病毒二重 RT-PCR 检测方法的建立及应用[J].中国畜牧兽医,2016,43(9):2455-2460,6.基金项目
河南省科技攻关项目(142102310224) （142102310224）
