我国首次牛结节性皮肤病病毒的分离鉴定  

Isolation and identification of lumpy skin disease virus from the first outbreak in China

作　　者：张敏敏[1] 孙亚杰 刘文兴[1] 刘任强[1] 王喜军[1] 步志高[1] ZHANG Min-min;SUN Ya-jie;LIU Wen-xing;LIU Ren-qiang;WANG Xi-jun;BU Zhi-gao(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China;College of Veterinary Medicine,Inner Mongolia Agricultural University Hohhot 01001&China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150069 [2]内蒙古农业大学兽医学院,内蒙古呼和浩特010018

出　　处：《中国预防兽医学报》2020年第10期1058-1061,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：“十三五”国家重点研发计划项目(2018YFC1200601);国家青年基金项目(31602074)。

摘　　要：牛结节性皮肤病(LSD)是由羊痘病毒属牛结节性皮肤病病毒(LSDV)引起的牛的烈性传染病。2019年8月,我国首次在新疆伊犁地区发现疑似LSD疫情。本研究采集伊犁疫情临床症状典型病牛病变皮肤组织,接种羊睾丸(LT)细胞,对出现典型细胞病变的培养物进行了系列鉴定。透射电镜观察到典型痘病毒形态的砖型粒子;山羊痘高免血清间接免疫荧光检测培养病变细胞显示阳性反应;LSDV基因组特异引物PCR检测结果为阳性;二代测序(De novo)全基因组分析显示与LSDV/Russia/Saratov/2017(MH646674.1)同源性最高(99.42%);系统进化分析显示与LSDV/Russia/Saratov/2017株相似,介于疫苗株和野毒株分支之间;病毒生长曲线测定显示感染LT细胞的生长特性与已报道的山羊痘病毒相似。上述结果表明,本实验分离得到引起我国首次LSD疫情的病毒株,命名为LSDV/China/Xinjiang/2019株(Xinjiang/2019),本研究为牛结节性皮肤病的防治奠定了基础。Lumpy skin disease(LSD)is a highly contagious cattle disease caused by lumpy skin disease virus(LSDV),belonging to the genus Capripoxvirus.In August 2019,an outbreak of suspected LSD occurred in the Yili area of Xinjiang autonomous region.In this study,skin samples were collected from the cattle with LSD typical signs for virus isolation.Cytopathic effects(CPE)were observed after inoculation of lamb testis(LT)cells.Subsequently,the isolated virus was identified by different methods and the following results were obtained:Brick-shaped viral particles can be visualized by electron microscopy using negative staining.Both indirect immunofluorescence assay and PCR detection were positive for LSDV.In addition,next-generation sequencing(De Novo)analysis revealed that the isolate,designated as LSDV/China/Xinjiang/2019(Xinjiang/2019),shared the highest homology with LSDV/Russia/Saratov/2017(MH646674.1)(99.42%).Phylogenetic analysis showed that the LSDV/China/Xinjiang/2019 clusters between the LSDV vaccine strain and the field strain LSDV Russia/Saratov/2017.Besides,the growth kinetics of the LSDV in LT cells was similar to that of GTPV.Taken together,we isolated the LSDV Xinjiang/2019 strain that was responsible in China,which is a basic work for the prevention and control of LSD.

关 键 词：牛结节性皮肤病病毒 分离鉴定 进化分析 

分 类 号：S857.5[农业科学—临床兽医学]