猪瘟病毒猪细小病毒猪繁殖与呼吸综合征病毒猪伪狂犬病病毒多重PCR方法的建立以及初步应用

猪瘟病毒、猪细小病毒、猪繁殖与呼吸综合征病毒及猪伪狂犬病病毒均能导致猪繁殖障碍,对养猪生产影响很大。本研究通过设计4对针对这4种病毒的特异引物建立了多重PCR方法,分别对其最佳反应条件、特异性及敏感性进行了测定,结果表明:其敏感性可达到CSFV 10 TCID50,PPV 10TCID50,PRRSV 1 TCID50,PRV 100 TCID50 CSFV 10TCID50,PPV 10TCID50,PRRSV 1 TCID50,PRV 100 TCID50。同时具有较好的特异性,对猪瘟病毒石门株、猪瘟病毒兔化弱毒株、PRV闽南A株、PPV弱毒疫苗株、PRRSV KY 35株及PRRSV B13株6个毒株均能扩增出相应的片段,而BVDV、BDV均未扩增出相应的片段。本方法的建立对于这4种病毒病的早期快速检测具有十分重要的意义。

Establishment and application of the multiplex PCR to the detection of CSF, PP, PRRS and AD viruses

classical swine fever virus(CSFV),porcine parvoviruses(PPV),porcine reproductive and respiratory syndrome virus(PRRSV) and Aujeszky's disease virus(ADV) all can result in reproductive failure of swine,which causes significant economical losses to the swine industry on a world-wide scale.Four primers were designed and the multiplex PCR were developed.Its sensitivity were CSFV 10 TCID50,PPV 10 TCID50,PRRSV 1 TCID50,PRV 100 TCID50 CSFV 10 TCID50,PPV 10 TCID50,PRRSV 1 TCID50,PRV 100 TCID50.The multiplex PCR for CSFV Shimen strain,CSFV lapinized virus strain,PRV Minnan A strain,PPV,PRRSV ky35 strain and B13 strain were done by this method.The DNA fragment were obtained from the six viral strains but not from BVDV and BDV.These results showed that the development of the multiplex PCR is very important for early and fast diagnosis of these diseases in china.