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

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.