牛病毒性腹泻病毒基因2型毒株的分离与鉴定

ISOLATION AND IDENTIFICATION OF BOVINE VIRAL DIARRHEA VIRUS-2 FROM THE ESTABLISED MDCK CELL LINE

本研究采用特异性RT-PCR从某实验室送检MDBK细胞中分离到1株牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV),命名为BVDV-C1。通过免疫荧光、电镜观察和5'UTR序列测定及分子进化分析,结果表明其在MDBK细胞中无细胞病变产生,应用牛病毒性腹泻病毒2型特异性单克隆抗体检测,在感染细胞胞浆内呈现特异性荧光信号,而牛病毒性腹泻病毒1型特异性单克隆抗体未检测到荧光。电镜观察病毒粒子呈圆形,有囊膜,直径约为50 nm。5'UTR序列分析表明该毒株属于BVDV-2b基因亚型。BVDV-C1株的分离对进一步开展疫苗研发、流行病学调查以及致病机理等方面的研究具有重要意义。

Bovine viral diarrhea virus 2 was isolated from the suspected contaminative MDBK cells. The virus isolate replicated in MDBK cells but did not induce cytopathogenic effect after five passages. Eventually, the virus isolate was identified in specific indirect immunofluorescent assay, RT-PCR, electron microscopy and was designated as BVDV-C1. The MDBK cells infected with BVDV-C1 showed strong fluorescent signal with BVDV-2 antibody. DNA fragment was amplified in RT-PCR from 5＇-UTR of the BVDV-C 1. The homology comparison and phylogenetic analysis showed that it belonged to BVDV-2b. Under electron microscopy, virus particles were near round and enveloped with size of 50 nm in diameter. The isolation of BVDV-C1 provided a basis for further research on vaccination, epidemiology and pathogenecity of BVDV in China.