犬冠状病毒Ⅱ毒株的分离鉴定及其基因的遗传进化分析  

Isolation and Identification of Canine CoronavirusⅡStrain and Genetic Evolution

作　　者：刘可欣 谭斌[1] 张傲 王倩颖 杨森 张淑琴[1] LIU Kexin;TAN Bin;ZHANG Ao;WANG Qianying;YANG Sen;ZHANG Shuqin(Institute of Special Animals and Plants,CAAS,Changchun 130122,China)

机构地区：[1]中国农业科学院特产研究所,长春130112

出　　处：《中国动物传染病学报》2024年第4期104-110,共7页Chinese Journal of Animal Infectious Diseases

基　　金：吉林省重点研发项目(20210202044NC)。

摘　　要：自长春市某宠物医院临床症状为肠胃炎的幼犬腹泻粪便中分离到1株病毒,该分离毒株在CRFK细胞上出现明显的细胞病变,经RT-PCR、序列测定和直接免疫荧光鉴定可知该分离毒株为犬冠状病毒株(Canine coronavirus,CCV),命名为CCV-CC01。利用PCR技术扩增CCV-CC01 S基因序列全长4273 bp,ORF-3序列1071 bp,N基因序列全长1202 bp。对分离毒株与其他冠状病毒毒株进行同源性比对并绘制系统进化树,结果显示该毒株与美国分离的CCV S378株亲缘关系最近,处于同一分支,与SARS-CoV-2亲缘关系较远,该毒株的出现可能与国际贸易相关。该毒株的分离鉴定为犬冠状病毒病的诊断、防治及后续疫苗研发奠定了基础。The fecal sample was collected for virus isolation from a diarrhea pup in a pet hospital in Changchun City.The isolated virus developed obvious cytopathic changes on CRFK cells.Then the virus isolate was identified to be Canine coronavirus(CCV)by RT-PCR,sequencing and direct immunofluorescence and designated as CCV-CC01.Subsequently,the S gene of the isolate CCV-CC01 was sequenced with the full-length of 4273 bp,ORF-31071 bp,N gene 1202 bp.The homology analysis of the S gene of this isolate and other coronavirus strains found that it was most closely related to the CCV S378 strain from the United States as both were in the same branch and far from CB/05,indicating its emergence might be related to international trade.The isolation and identification of the CCV-CC01 strain laid the foundation for the diagnosis,prevention and vaccine development of canine coronavirus disease.

关 键 词：CCV 分离鉴定 直接免疫荧光法 遗传进化分析 

分 类 号：S852.65[农业科学—基础兽医学]