用PCR技术鉴定犬细小病毒弱毒疫苗株

Establishment of Polymerase Chain Reaction for Discrimination of Vaccine Strains of Canine Parvovirus

通过分析比较犬细小病毒 (CPV)弱毒疫苗株和强毒株的基因序列 ,设计了 3条引物并组成 2对引物 ,对弱毒疫苗株和强毒株进行了半嵌套式 PCR扩增。第 1轮 PCR扩增的结果为 CPV弱毒疫苗株和强毒株有相同的目的片段(5 85 bp) ;第 2轮 PCR扩增的结果为 CPV强毒株有 375 bp目的片段 ,而 CPV弱毒疫苗株没有 375 bp目的片段。对PCR产物进行电泳和酶切分析 ,结果证明 PCR产物片段大小和酶切位点与设计的产物完全一致。特异性和敏感性测定结果表明 ,该方法是高度特异和敏感的 。

Two pairs of PCR primers were designed according to analyzing and contrasting the sequances of the vaccine strain and virulent strain of CPV,a heminested PCR method was established.Result of the first PCR amplification showed the same amplified products of 585 bp length,after the second PCR amplification,the virulent strain produced the length 375 bp fragment,but the vaccine strain couldn′t .The products of PCR were examined by electrophoresis and restriction enzyme digestion.The result showed the length of the fragment and enzyme sites were as the same as those designed.The PCR assay of CPV was proved to be specific and sensitive.It shows that this method may be used for discriminating the vaccine strain and virulent strain of CPV or monitoring the vaccinated canine.