水貂肠炎细小病毒的分离鉴定及其VP2基因的序列分析

本试验从疑似细小病毒感染的病死水貂中分离到1株病毒。经PCR鉴定、细胞培养、蛋白质电泳鉴定最终确定为水貂肠炎细小病毒(MEV),命名为MEV-WFD。对该分离病毒的衣壳蛋白VP2基因进行克隆测序分析,结果表明此病毒VP2基因3处碱基发生点突变,其中一处的突变导致第328处氨基酸残基由疏水性丙氨酸(Ala)变为亲水性苏氨酸(Thr)。将此株病毒VP2基因与GenBank上公布的所有MEV的VP2基因碱基序列进行同源性比较及进化树分析,结果表明该病毒与ZYL-1、MEV/LN/-10和Manzhouli的VP2基因同源率最高,为99.8%;进化树构建结果表明,该病毒与6株已公布的病毒属于同一进化分支。

Isolation,Identification of Mink Enteritis Virus and Sequence Analysis of its VP2 Gene

A virus was isolated from a dead mink which was suspected of being infected with the mink enteritis virus(MEV). It was eventually identified as MEV using PCR assay, cell culture and SDS-PAGE, and named MEV-WFD. The VP2 gene of this parvovirus isolate was cloned into pMD18-T vector, sequenced and analyzed. As a result,the VP2 protein possessed one sbustitution, 328A-T.The result of the nucleotide homologies of VP2 gene between this isolate and all MEV published on GenBank illustrated that MEV-WFD had the the highest homology 99.8% with ZYL-1,MEV/LN/-10 and Manzhouli.VP2 gene phylogenetic anaysis showed that this isolate and other six wild strains had the same ancestor.This research had laid the foundation for the mink enteritis parvovirus molecular epidemiological investigation.