一种鉴别猪瘟C-株和其他毒株的分子生物学方法

根据已发表的猪瘟病毒(CSFV)核苷酸序列,筛选出C-株与其他毒株的核苷酸序列差异标记CTTTTTTCTTTT,并设计包含此标记序列的CSFV特异性PCR引物对及nested-PCR引物对。从待检猪脾脏或淋巴组织中提取总RNA,用AMV逆转录酶进行逆转录后,进行PCR和nested-PCR。将扩增片段纯化回收并克隆到pMD18-T载体进行测序。对测序结果进行分析,含有此标记序列的即为C-株,不含此标记序列的则为其他毒株。

A molecular biology method to differentiate C-strain from other strains of CSFV

A difference marker sequence CTTTTTTCTTTT was selected from C-strain and other strains of CSFV,and specific PCR primer pair and nested-PCR primer pair were designed at the basis of previously published sequences of CSFV.Total RNA was extracted from spleens or lymph nodes of suspicious ill pigs.Target fragments were amplified utilizing RT-PCR and nested-PCR techniques.After purification and recovery,the target fragments were cloned into pMD18-T vectors and further sequenced.According to sequence analysis,sequence containing the marker sequence belongs to C-strain and others belongs to other strains of CSFV.