猪细小病毒和猪源脑心肌炎病毒的二重PCR检测方法的建立

Establishment of a Diplex PCR Assay for Detection of PPV and Porcine EMCV

试验旨在建立能同时检测猪细小病毒(porcine parvovirus,PPV)和猪源脑心肌炎病毒(encephalomyocarditis virus,EM-CV)的二重PCR检测方法。根据GenBank中PPV VP2基因和EMCV 3D基因序列,利用Premier 5.0软件设计出PPV和EMCV的特异性引物。通过优化反应条件,建立了能同时特异性检测PPV和EMCV的二重PCR方法,PPV和EMCV的敏感性检测极限分别达7.62和1.22×10-1pg/μL。通过对临床36份样品的检测结果表明,该二重PCR检测方法敏感、特异,适合于临床检测应用。

Establish a diplex PCR assay was to detect porcine parvovirus（PPV） and porcine encephalo myocarditis virus（EMCV） simultaneously.According to PPV VP2 gene sequences and EMCV 3D gene sequences in GenBank,special primers of PPV and EMCV were designed by Premier 5.0 software.Trough optimizing the reaction conditions,a diplex PCR assay,which could detect PPV and EMCV simultaneously,was established.The sensitivity of PPV and EMCV were 7.62 and 1.22×10-1 pg/μL respectively.Clinical tests of 36 samples showed that the diplex PCR detection assay was sensitive,specific,and suitable for clinical application.