牛轮状病毒的分离及其RT-PCR鉴定  

Isolation of Bovine Rotavirus and Identification by Using RT-PCR

作　　者：姜向阳[1] 邓小敏[1] 王晶钰[1] 王学艳[1] 罗艳[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《西北农业学报》2007年第4期191-194,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：陕西省科技攻关项目(2005K01-G20-02)

摘　　要：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 .

关 键 词：轮状病毒 细胞培养 VPl基因 MAl04细胞 

分 类 号：S852.65[农业科学—基础兽医学]