PRRSV普通株与变异株鉴别诊断RT-PCR方法的建立与临床应用  

Establishment of differential diagnostic method for normal and mutant PRRSV strains by RT-PCR

作　　者：周丹[1,2] 郭万柱[1,3] 漆信桥[1] 韩国全[4] 徐志文[1] 朱玲[1] 

机构地区：[1]四川农业大学动物生物技术中心,四川雅安625014 [2]中牧股份成都药械厂,四川雅安610014 [3]四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安625014 [4]四川农业大学动物营养研究所,四川雅安625014

出　　处：《中国兽医学报》2012年第3期367-370,377,共5页Chinese Journal of Veterinary Science

基　　金：教育部长江学者和创新团队发展计划资助项目(IR-TO848)

摘　　要：通过对GenBank已公布猪繁殖与呼吸综合征病毒(PRRSV)普通株与变异株Nsp2基因序列进行比对分析,设计合成3条引物,建立一步鉴别诊断普通株与变异株PRRSV的RT-PCR检测方法。结果表明,PCR扩增获得的普通株片段大小为267bp,变异株片段大小为180bp。敏感性试验结果表明该方法具有较高的敏感性,能检测到含量为1.5pg的核酸含量;特异性结果表明该方法在同等条件下检测PRV、CSFV、PCV-2、JEV、CPV、TGEV、SIV均为阴性。对2009-2010年送检的100份临床样品进行检测,结果检出26份变异PRRSV,8份普通PRRSV,阳性率为34%;与ELISA抗体检测试剂盒比较,多检出4份阳性样品。表明建立的鉴别诊断PRRSV的RT-PCR检测方法具有敏感性高、特异性强、重复性好的特点,适用于临床针对PRRSV的检测。Through analyze the Nsp2 gene of PRRSV published in GenBank nucleotide sequence database,3 primers were designed and one step detected RT-PCR method was established.The result showed that,180 bp specifically fragment and 267 bp fragment were amplified from variant PRRSV and classical form PRRSV.The developed method was sensitive to detect the RNA as low as 1.5 pg.The negative results were found on PRV,CSFV,PCV-2,JEV,CPV,TGEV,SIV.One hundred clinical specimens of sick pigs were tested by the method from 2009 to 2010,26 percent of them were variant PRRSV and 8 percent of them were classical form PRRSV.The results showed that the RT-PCR assay is high specificity,sensiticity and reproducibility,and could be used to differentiate common PRRSV.

关 键 词：PRRSV NSP2基因 RT-PCR 

分 类 号：S852.65[农业科学—基础兽医学]