猪伪狂犬病病毒和猪圆环病毒2型多重PCR检测方法的建立与初步应用

The development and preliminary application of multiplex PCR for detection of PRV and PCV-2

为了建立可同时检测猪伪狂犬病病毒(PRV)和猪圆环病毒2型(PCV-2)的多重PCR检测方法,本研究根据GenBank中登录的PRV gE基因和PCV-2 ORF2基因的核苷酸序列,分别设计了2对特异性引物PRV-S/PRV-A和PCV-2-S/PCV-2-A,产物分别为270 bp和189 bp。检测结果显示,该方法具有较高特异性,检测PRV和PCV-2的DNA最低量均为1×10^(-4)μg/μL。对136份临床上疑似为PRV和PCV-2感染病料样品进行检测的结果表明,本研究建立的多重PCR检测方法可用于临床上PRV和PCV-2引起的单纯或混合感染的传染病病原学诊断。

To establish the assay for detections of pseudorabies virus（PRV）and Porcine circovirus type 2（PCV-2）,a multiplex RT-PCR was developed with two pair specific primers of PRV-S/PRV-A and PCV-2-S/PCV-2-A designed for detecting PRV and PCV-2 based on the gE gene and ORF2 gene sequences in GenBank respectively.The results showed that the assay was able to detect PRV and PCV-2 simultaneously by amplifying DNA fragments of 270 bp and 189 bp,respectively.The sensitivity of the assay was 1×10-（-4） μg/μL of DNA.Our result of the 136 tested clinical sample suggest that the Multiplex PCR detection methods can be used to detect the,PRV and PCV-2.