牛病毒性腹泻病毒基因2型云南株的分离与鉴定  

Isolation and identification of Yunnan strain of bovine viral diarrhea virus-2

作　　者：普浩宇 亐开兴[2] 杨贵伟 程美玲[1] 黄必志[2] 王蓉 任珂青 尹革芬[1] PU Haoyu;QU Kaixing;YANG Guiwei;CHENG Meiling;HUANG Bizhi;WANG Rong;REN Keqing;YIN Gefen(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;Yunnan Academy of Grassland and Animal Science,Science,Kunming 650212,China)

机构地区：[1]云南农业大学动物医学学院,云南昆明650201 [2]云南省草地动物科学研究院,云南昆明650212

出　　处：《中国兽医学报》2021年第10期1922-1929,共8页Chinese Journal of Veterinary Science

基　　金：云南省高校科技创新团队支持计划资助项目(IRTSTYN);云南省后备人才培养计划基金资助项目(2018HB045);云南省重点研发专项基金资助项目(2019ZG011)。

摘　　要：本研究利用MDBK细胞分离牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV),通过IFA、qRT-PCR和ELISA方法鉴定,从云南分离获得了1株基因2型BVDV,命名为YNJG。采用IFA,按Reed-Muench法计算出YNJG分离株的滴度为10-4.4 TCID50/100μL。根据其5′UTR序列进行基因分型,确定YNJG属于BVDV-2毒株。全基因序列测序结果显示,YNJG毒株全长为12288个核苷酸,编码3898个氨基酸,GenBank登录号为MW168422,与美国BVDV分离株9231核苷酸同源性最高,为95.4%,该毒株不能导致细胞病变,属于非致细胞病变型毒株。毒力分析显示,该毒株5′UTR第216位和275位核苷酸分别为C和T,且1142位氨基酸处无氨基酸的插入,属于低毒力毒株。Bovine viral diarrhea virus(BVDV)is one of the most important pathogens that harms the development of cattle industry.In this study,MDBK cells were used to isolate virus,and the isolated virus was identified by IFA,qRT-PCR and ELISA methods.The results showed that a genotype 2 bovine viral diarrhea virus(BVDV)was isolated from Yunnan Province,it was named as YNJG isolate.Using immunofluorescence assay and Reed-Muench method,the titer for YNJG isolate was calculated as 10^(-4.4)TCID_(50)/100μL.Based on the 5′UTR sequence identity,YNJG isolate was grouped to BVDV-2 type.The whole virus genome was sequenced,it was showed that YNJG isolate was 12286 nucleotides(nt)in length,encoding 3898 amino acids(GenBank accession No.MW168422).The strain shared the highest sequence homology with the US isolate 9231(95.4%),it could not cause cytopathic effect(CPE)in MDBK cells,indicating that it was a noncytopathogenic(NCP)strain.The virulence analysis of the YNJG isolate showed that the nucleotides at positions 216 and 275 of its 5′UTR were C and T,respectively,and there was no extra amino acid insertion at position 1142 of the full amino acid sequence,suggesting it was likely a low-virulence strain.

关 键 词：牛病毒性腹泻病毒基因2型 YNJG分离株 5′UTR 遗传进化分析 

分 类 号：S852.65[农业科学—基础兽医学]