牛轮状病毒的分离及其RT-PCR鉴定

Isolation of Bovine Rotavirus and Identification by Using RT-PCR

采用MA104细胞从疑似轮状病毒腹泻犊牛粪样中进行轮状病毒分离,盲传4代后接种细胞出现明显细胞病变(CPE),对分离病毒进行核酸聚丙烯酰胺凝胶电泳,电泳条带为4∶2∶3∶2型,具备典型A群轮状病毒特征。依据NCBI等登录的牛轮状病毒的VP1基因序列设计一对引物,对其VP1基因进行克隆及序列分析,结果分离毒与国外牛轮状病毒分离株RF的核苷酸同源性为98.9%,与UK的核苷酸同源性为91.5%,证明分离毒株是一株牛轮状病毒。

One strain of bovine rotavirus was isolated from excrement of calf by using MA104 cell, and produced obvious cytopathic effects after fostering 4 times. RNA of isolated virus was analysed by PAGE and was type of 4 ： 2 ： 3 ： 2 , which belonged to rotavirus A type. A pair of primers were designed to clone VP1 gene according to sequences of bovine rotavirus in NCBI. The results showed that nucleotides sequence of VP1 shared 98.9% with RF isolated strain , 91.5% with UK .