牛冠状病毒的分离鉴定

Isolation and Identification of Bovine Coronavirus

将腹泻犊牛粪样上清接种于HCT-8细胞,出现CPE后收集病毒。提取病毒RNA后,巢氏反转录PCR扩增出冠状病毒N基因全长1437bp和N基因内部355bp片断,基因测序结果与Mebus株冠状病毒的同源性在98.3%,结果表明从腹泻的新生犊牛粪样中分离出的病毒确系牛冠状病毒。

The fecal suspension of a diarrheic calf was inoculated into HCT-8 cells, harvesting the virus when the CPE appeared. Virus RNA was extracted, and the Nested RT-PCR was performed to amplify the entire N genes with the length of 1437bp and a 355bp fragment of N gene in coronavirus. The homology comparison of the N genes sequence with Mebus strain was 98.3%. The result showed that a bovine coronavirus strain was isolated from diarrheic feces of a newborn calf.