PCV2-PRV-PRRSV-CSFV多重PCR检测方法的建立  

Establishment of multiplex PCR for simultaneous detection of porcine circovirus type 2,pseudorabies virus,porcine reproductive and respiratory syndrome virus and classical swine fever virus

作　　者：左奕[1] 王建永[1] 袁万哲[1] 孙继国[1] 

机构地区：[1]河北农业大学动物医学院/河北省兽医生物技术工程技术研究中心,河北保定071001

出　　处：《中国兽医科学》2015年第8期771-775,共5页Chinese Veterinary Science

基　　金：2015年度河北省畜牧兽医局科技项目(2015-01-04)

摘　　要：根据GenBank中已发表的猪圆环病毒2型(PCV2)、伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)的有关基因序列,设计4对引物分别用于扩增PCV2的ORF2基因、PRV的gH基因、PRRSV的ORF7基因和CSFV的5′UTR基因的目的片段,建立了同时检测4种病毒的多重PCR。敏感性和特异性试验结果表明,该方法对这4种病毒的最低核酸检出量分别为48.1pg(PCV2)、29.6pg(PRV)、54.2pg(PRRSV)和62.8pg(CSFV)。该方法可同时扩增440bp(PCV2)、359bp(PRV)、321bp(PRRSV)和186bp(CSFV)的特异性片段,而对牛流行性腹泻病毒、猪流行性腹泻病毒和猪细小病毒的检测结果均为阴性。多重PCR与单一PCR的符合率为100%。结果表明,该方法可用于这4种病毒的临床快速检测与流行病学调查。A multiplex PCR was developed using four pairs of primers designed according to the published relative sequences of porcine circovirus type 2（PCV2）, pseudorabies virus（PRV）,porcine reproductive and respiratory syndrome virus（PRRSV） and classical swine fever virus（CSFV）, respectively. Based on the similarity of the viral sequences deposited in the GenBank database, the ORF2 gene of PCV2, the gH gene of PRV, the ORF7 gene of PRRSV and the 5＇UTR gene of CSFV were selected as the diagnostic targets. Specific PCR products of 440 bp for PCV2,359 bp for PRV,321 bp for PRRSV and 186 bp for CSFV could be simultaneously amplified by the multiplex PCR. By comparison,the multiplex PCR and the single PCRs showed 100% coincidence. In conclusion,the multiplex PCR could be used for the detection and differential diagnosis of those four virus.

关 键 词：猪圆环病毒2型 伪狂犬病病毒 猪繁殖与呼吸综合征病毒 猪瘟病毒 多重PCR 

分 类 号：S852.651[农业科学—基础兽医学]