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草鱼呼肠孤病毒JX-0902株的分离和鉴定
引用本文:刘永奎,王庆,曾伟伟,石存斌,张超,陈道印,吴淑勤.草鱼呼肠孤病毒JX-0902株的分离和鉴定[J].中国水产科学,2011,18(5):1077-1083.
作者姓名:刘永奎  王庆  曾伟伟  石存斌  张超  陈道印  吴淑勤
作者单位:中国水产科学研究院珠江水产研究所;上海海洋大学;济宁市渔业监测站;南昌县莲塘鱼病防治所;
基金项目:农业部公益性行业科研专项(200803013); 现代农业产业技术体系建设专项资金(nycytx-49-14)
摘    要:收集江西南昌地区患典型草鱼出血病的病鱼组织,进行超薄切片电镜观察,结果显示,在脾、肾样品中发现大量病毒颗粒,病毒无囊膜,近球形,直径约70 nm,形态与已报道的草鱼呼肠孤病毒(grass carp reovirus,GCRV)相似。取病鱼肌肉、内脏研磨,组织液经离心、过滤除菌后进行鱼体人工感染试验和细胞感染实验,结果发现,人工感染试验鱼出现死亡,且具有典型出血症状;组织滤液感染草鱼肾细胞系(CIK)细胞后,盲传6代未观察到典型细胞病变效应,但感染细胞固定后经超薄切片电镜观察,细胞质内有大量病毒存在,与病鱼组织切片观察到的病毒形态一致。SDS-PAGE结果进一步揭示,新分离病毒株(暂命名JX-0902)基因组由11条dsRNA组成,呈现水生呼肠孤病毒基因组典型特征,但基因组带型与GCRV 873株存在差异,而与HZ08株接近。根据GenBank上已提交的草鱼呼肠孤病毒S6序列(GQ896337)设计特异引物,以JX-0902株的cDNA作为模板,可扩增出特异性条带。基于S6序列的聚类分析结果显示,JX-0902与GCRV HZ08株、GD108株S6同源性高达98%、99%,该分离株应为草鱼呼肠孤病毒。

关 键 词:草鱼出血病  呼肠孤病毒  江西分离株  分离  鉴定  
修稿时间:2011/10/8 0:00:00

Isolation and identification of grass carp reovirus strain JX-0902
LIU Yongkui,WANG Qing,ZENG Weiwei,SHI Cunbin,ZHANG Chao,CHEN Daoyin,WU Shuqin.Isolation and identification of grass carp reovirus strain JX-0902[J].Journal of Fishery Sciences of China,2011,18(5):1077-1083.
Authors:LIU Yongkui  WANG Qing  ZENG Weiwei  SHI Cunbin  ZHANG Chao  CHEN Daoyin  WU Shuqin
Institution:LIU Yongkui1,2,WANG Qing1,ZENG Weiwei1,SHI Cunbin1,ZHANG Chao3,CHEN Daoyin4,WU Shuqin1 1.Pearl River Fisheries Research Institute of Chinese Academy of Fishery Sciences,Guangzhou 510380,China 2.Shanghai Ocean University,Shanghai 201306,China 3.Jining Fishery Monitoring Center,Jining 272019,China 4.Fish Disease Control Center in Liantang Town of Nanchang,Nanchang 330200,China
Abstract:We identified a new strain of reovirus in grass carp from Nanchang city,Jiangxi Province.We collected diseased yearling grass carp,Ctenopharyngodon idellus exhibiting typical hemorrhage symptoms from Nanchang city and observed large numbers of viral cells in ultra-thin sections of the spleen and kidney.The virus had a non-enveloped structure with a diameter of approximately 70 nm,very similar to the characteristics of the grass carp reovirus(GCRV).We isolated samples of the virus by homogenizing the tissue,...
Keywords:grass carp hemorrhage  reovirus  Jiangxi isolate  separation  identification  
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