2株牛病毒性腹泻病毒的分离鉴定  

Isolation and Identification of Two Bovine Viral Diarrhea Virus Strains

作　　者：秦义娴 陈晓春[1] 刘丹[1] 孙淼[1] 薛青红[1] 吴华伟[1] 高金源[1] 高月异 QIN Yi-xian;CHEN Xiao-chun;LIU Dan;SUN Miao;XUE Qing-hong;WU Hua-wei;GAO Jin-yuan;GAO Yue-yi(China Institute of Veterinary Drug Control,Beijing,100081,China)

机构地区：[1]中国兽医药品监察所,北京100081

出　　处：《动物医学进展》2023年第1期43-47,共5页Progress In Veterinary Medicine

基　　金：中国兽医药品监察所“兽药行业公益性重点专项”项目(GY202103);国家重点研发计划项目(2022YFD1800600)。

摘　　要：牛病毒性腹泻是目前危害我国养牛业发展的重要传染病之一,在我国多个省、自治区、直辖市均有流行。对2头疑似感染牛病毒性腹泻病毒的牛血清样本进行检测、分离病原,并根据E2基因序列确定分离株的基因亚型。将2份血清样本经RT-PCR检测为阳性后,接种MDBK细胞,通过连续传代培养并进行间接免疫荧光测定,成功分离到2株非致细胞病变型(NCP)BVDV分离毒株,分别命名为HN和XJ-2。对2个毒株的E2基因进行克隆及序列测定,并与NCBI上登录的BVDV参考毒株E2基因序列进行同源性比较和遗传进化分析。结果表明,XJ-2株、HN株与BVDV1b亚型JL^(-1)株处于同一分支,核苷酸同源性分别为99.4%和99.3%,均属于BVDV1b亚型。对新疆、河南两地XJ-2和HN株的分离鉴定,可为我国BVDV的分子流行病学研究和防控提供数据参考。Bovine viral diarrhea is one of the important infectious diseases endangering the development of cattle industry in China,and it is prevalent in many provinces,cities and autonomous regions.In this study,2 bovine serum samples suspected of being infected with bovine viral diarrhea virus were tested and the pathogen was isolated,and the genotype of the isolates was determined according to E2 gene sequence.After the two serum samples were detected as positive by RT-PCR,MDBK cells were inoculated.Through continuous subculture and indirect immunofluorescence assay,two non cytopathic(NCP)BVDV isolates were successfully isolated,named HN and XJ-2 respectively.The E2 genes of two strains were cloned and sequenced,and compared with theE2 gene sequences of BVDV reference strains registered in NCBI.The results showed that XJ-2 strain,HN strain and BVDV1b subtype JL^(-1)strain were in the same branch,and the nucleotide homology was 99.4%and 99.3%respectively.The isolation and identification of XJ-2 and HN strains in Xinjiang and Henan provide data reference for the molecular epidemiological research and prevention and control of BVDV in China.

关 键 词：牛病毒性腹泻病毒 分离鉴定 同源性比较 遗传进化分析 

分 类 号：S852.659.6[农业科学—基础兽医学] S858.23[农业科学—兽医学]