中原地区牛肠道病毒分离鉴定及其遗传变异分析  

Isolation,identification and genetic evolution analysis of bovine enterovirus

作　　者：刘颖[1] 黄宗梅[1] 王轩昂 郑兰兰[1] 马世杰[1] 陈红英[1] LIU Ying;HUANG Zongmei;WANG Xuanang;ZHENG Lanlan;MA Shijie;CHEN Hongying(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046

出　　处：《畜牧与兽医》2025年第5期88-92,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划项目(2022YFD1601606);河南省高校科技创新人才支持计划项目(21HASTIT039)。

摘　　要：旨在探究中原地区牛肠道病毒(BEV)的生物学特性及其5′-UTR序列遗传特征。采用RT-PCR方法对2023—2024年从四川、河南和河北地区收集的77份腹泻牛粪便样品进行BEV检测,对阳性样本的BEV 5′-UTR序列进行克隆及遗传变异分析,通过接种Madin-Darby牛肾细胞(MDBK)进行病毒分离与鉴定。结果:77份牛粪便样品中有20份(25.97%)检测为BEV阳性,获得20株BEV 5′-UTR核苷酸序列;20株BEV与13株BEV参考毒株5′-UTR序列同源性为77.2%~94.1%;遗传变异分析显示,获得的20株BEV序列中17株属于BEV-F种,余下3株属于BEV-E种;分离到2株BEV,命名为HB002、HN004-8,均为BEV-F种,病毒滴度均在48 h时达到高峰,分别为10^(5.49)TCID_(50)/μL和10^(5.67)TCID_(50)/μL;电子显微镜观察病毒粒子呈球形,直径20~30 nm。本研究为BEV感染的防控提供了参考。To understand the biological properties and genetic characteristics of bovine enterovirus(BEV)in central China,a total of 77 fecal samples were collected from cattle with diarrhoea in the area in 2023-2024 to detect BEV by RT-PCR.The BEV-positive samples were subjected to the cloning of 5′-UTR sequences and the genetic variation analysis,and was inoculated into Madin-Darby bovine kidney cells(MDBK)for isolation and characterization of BEV.The results showed that 20 of the 77(25.97%)bovine diarrhea samples tested positive for BEV,and the 5′-UTR nucleotide sequences of 20 BEV strains were obtained.The 5′-UTR nucleotide sequence identity of the 20 BEV strains obtained in this study and the 13 BEV reference strains ranged from 77.2%to 94.1%.The phylogenetic analysis showed that 17 of the 20 BEV strains obtained in this study belonged to the BEV-F type,while the other 3 strains belonged to the BEV-E type.Two BEV strains(HB002 and HN004-8)were isolated,both of which were BEV-F type,with viral titers of 10^(5.49)TCID_(50)/μL and 10^(5.67)TCID_(50)/μL,respectively,and reached the peak of viral titer at 48 h.The results of electron microscopy showed that the viral particles were round and 20-30 nm in diameter.This study provided a reference for comprehensive prevention and control of BEV infection.

关 键 词：牛肠道病毒 分离与鉴定 5′-UTR 遗传变异 

分 类 号：S855[农业科学—临床兽医学]