1株感染洛氏鱥呼肠孤病毒的分离与培养  

Isolation and culture of one reovirus strain in Phoxinus lagowskii Dybowskii

作　　者：巴翠玉 张培军[2] 赵福广[1] 张林波[1] 焦雪[1] 李月红[1] 

机构地区：[1]吉林农业大学,吉林长春130118 [2]吉林省卫生监测检验中心,吉林长春130000

出　　处：《中国兽医学报》2018年第1期64-68,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972191);农业部948资助项目(2014Z34);吉林省留学人员创新创业资助项目(201523)

摘　　要：利用细胞培养技术从养殖发病洛氏鲅中分离出1株病毒，经人工感染试验、电镜观察、核酸基因组SDS-PAGE电泳、RT—PCR检测确认为呼肠孤病毒。人工感染发病的症状与自然发病症状相同；病毒感染可使鲤鱼上皮瘤细胞（EPC）产生典型细胞病变效应，收集细胞上清，负染后电镜下可观察到大小均一，直径约70nm，近球形，无囊膜结构的病毒粒子，初步判断为呼肠孤病毒；病毒基因组SDS-PAGE结果表明，病毒基因组由11个分段的RNA核酸片段组成，为典型的水生呼肠孤病毒核酸带型。病毒基因纽$10节段RT-PCR结果表明，有特异性条带，其与GenBank中登录的呼肠孤病毒S10节段（JN206665．1）序列同源性为91％，该分离株应为呼肠孤病毒，本研究首次在洛氏缄鱼内分离到呼肠孤病毒。A virus strain was isolated from the onset of breeding Rockwell in Phoxinus lagowskii Dybowskii by cell culture technology. After artificial infection in fish,electron microscopy observation,genomic SDS-PAGE, RT-PCR, the virus strain was identified as reovirus. The same symptoms as the natural was reproduced by artificial infection. Virus infection in carp epithelial tumor cells （EPC） could produce typical cytopathic effect, supernatant was collected to make negative staining for electron microscope,a diameter of about 70 nm, subglobose,no capsular structure of the virus particle was founded,which got a preliminary judge for reovirus ;the virus genome SDS- PAGE results showed that the viral genome was composed of 11 segments of RNA nucleic acid fragments,which were typical of the viral genome. Viral genome S10 segment RT-PCR results showed 91% homology with the reovirus S10 segment JN206665.1 in GenBank. The reovirus was first separated in the Rockwell mandarin fish in our research.

关 键 词：洛氏鱥 呼肠孤病毒 分离与培养 

分 类 号：S852.65[农业科学—基础兽医学]