蓝舌病病毒7型毒株的分离与分子鉴定  

Isolation and Identification of Bluetongue Virus Serotype 7 Field Isolate

作　　者：吕敏娜[1] 杨恒[2] 孙铭飞[1] 李娟[1] 林栩慧 张健騑[1] 廖申权[1] 戚南山[1] 吴彩艳[1] 李华春 陈琴苓[1] 

机构地区：[1]广东省农业科学院动物卫生研究所广东省兽医公共卫生公共实验室广东省畜禽疫病防治研究重点实验室,广州510640 [2]云南省畜牧兽医科学院,昆明650224

出　　处：《畜牧兽医学报》2017年第7期1281-1287,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：公益性行业(农业)科研专项(201303035);广东省科技计划项目(2015A020209075;2015A020209084)

摘　　要：为监测广东省蓝舌病流行状况,2014年从广东省某一奶牛场采集血液样品,检测虫媒病病原,对经RTPCR检测蓝舌病病毒核酸阳性的样品进行病毒分离并鉴定。经鸡胚静脉接种后取肝,研磨后离心取上清转接C6/36细胞和BHK-21细胞,出现CPE,RT-PCR检测蓝舌病病毒的VP7基因并进行序列比对,电镜观察病毒粒子,证实分离到蓝舌病病毒,命名为GD/ST2014。进一步扩增VP2基因,进行系统发育树分析,以及血清中和试验,表明此分离株为血清型7型。这是国内首次报道牛源蓝舌病病毒血清型7型的分离鉴定。In 2014,cattle blood samples were collected from Guangdong province and vector-borne pathogens contained in them were detected in the present study.Samples which were detected as BTV positive by quantitative reverse transcription polymerase chain reaction（qRT-PCR）were isolated and identified.Embryonated chicken eggs were subsequently inoculated and their livers were harvested to passage on C6/36 and BHK-21 cells,respectively.Extensive cytopathic changes were observed.A subsequent qRT-PCR assay target with VP7 gene and SEM observation results further confirmed the presence of BTV,and named as GD/ST2014.In addition,VP2 gene fragment was used for genotyping.And combined with VNT test results,the genotype of the isolate obtained in the present study were identified as BTV-7.This was the first report that bluetongue virus serotype 7 field isolate was isolated from cattle in China.

关 键 词：蓝舌病病毒 分离 鉴定 血清型 序列分析 

分 类 号：S852.659.4[农业科学—基础兽医学]