广东一株牛源流行性出血病病毒的分离鉴定  

Isolation and identification of the epizootic hemorrhagic disease virus from cattle in Guangdong

作　　者：吕敏娜[1] 朱建波 李娟[1] 杨振兴 林栩慧 陈琴苓[1] 张健騑[1] 廖申权[1] 戚南山[1] 吴彩艳[1] 陈志虹[1] 李华春 孙铭飞[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,广东广州510640 [2]云南省畜牧兽医科学院,云南昆明650224

出　　处：《中国预防兽医学报》2017年第1期67-70,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：公益性行业(农业)科研专项(201303035);广东省科技计划项目(2015A020209084);广东省科技计划项目(2015A020209075);广东省科技计划项目(2015A020209073)

摘　　要：为了解广东反刍动物流行性出血病(EHD)的感染情况,本研究对采集的牛血液样品进行EHD病毒(EHDV)的RT-PCR检测,对核酸检测为阳性的样品进行病毒分离鉴定。将阳性样品接种BHK-21细胞连续传3代时出现细胞病变,扩增出了VP7基因,进行BLAST序列比对,结果表明该分离株与国内外EHDV分离株的同源性达98%~100%,TCID_(50)为10^(-6.5)/50μL,不能凝集鸡红细胞,表明该分离株为EHDV,命名为GD-N133。根据VP2基因序列的分析和微量中和试验结果,确定该病毒株为EHDV-1型。To investigate the prevalence of epizoodc hemorrhagic disease virus （EHDV） among ruminants in Guangdong province, the cattle blood samples were collected and detected by RT-PCR, then the positive samples were inoculated into BHK-21 cells for the virus isolation, and a virus was isolated. In addition, VP7 gene fragment was amplified by RT-PCR and sequenced. The alignment of the VP7 sequence showed that the isolate was high similarity （98%-100%） with other EHDV reference strains available in GenBank. The virus titer was 10^-6.5 TCID50 in BHK-21 cells, and unable to agglutinate chicken red blood cells. All the results indicated that the isolate was EHDV, named GD-N133. Furthermore, combined with virus neutralization test and the VP2 gene fragment analysis for genotyping the virus, the genotype of the isolate was identified as EHDV-1.

关 键 词：流行性出血病病毒 牛 血清型 分离 鉴定 

分 类 号：S852.65[农业科学—基础兽医学]