作　　者： ; ; ; ; ; ; ;

机构地区： 中国水产科学研究院珠江水产研究所

出　　处： 《中国水产科学》 2010年第6期1257-1263,共7页

摘　　要： 从浙江省湖州地区采集发病草鱼(Ctenopharyngodon idellus)样本中,取症状明显病料的肝、脾、肾组织,经过滤除菌处理后接种草鱼肾脏细胞(CIK)。盲传8代,CIK细胞未出现明显细胞病变,但感染细胞固定后经电镜观察发现,细胞质内有大量病毒聚集,病毒无囊膜,近球形,直径约70nm,形态与已报道的草鱼呼肠孤病毒(Grass Carp Reovirus,GCRV)相似。将病毒提纯后分别经DNA酶、RNA酶和绿豆核酸酶消化,证实为双链RNA(dsRNA)病毒。十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结果显示,病毒基因组由11个dsRNA组成,呈现水生呼肠孤病毒基因组典型特征。采用RNA水平3'末端加接头的方法获得了S6节段的全长序列,测序结果表明,S6由2030个核苷酸组成,推测其编码一个分子量约为68.4kD的蛋白。聚类分析结果显示,该病毒为水生呼肠孤病毒,但在氨基酸水平上与草鱼呼肠孤病毒代表株873株的差异较大,同源性为33%,提示该病毒为一株新型的草鱼呼肠孤病毒。本研究旨在为草鱼出血病防治方法的深入研究奠定基础。 Diseased Grass carp（Ctenopharyngodon idellus） were collected from Huzhou,Zhejiang Province.After filtration treatment,the tissue suspension was inoculated in CIK ce1ls,but no obvious cytopathic effects（CPE） were observed even after passages of eight times.However,plenty of nonenveloped virus particles with subsphaeroidal shape were observed under electron microscope（EM）,which was highly similar to reported grass carp reovirus（GCRV）.The nucleic acid of purified virus was digested by DNases,RNases and Mung-bean Nuclease respectively.The results showed the nucletic acid of the virus was double-stranded RNA,and the SDS-PAGE electropherosis further suggested the virus possessed 11 segments of dsRNA,which was the typical characteristic of GCRV.In order to classify the strain on the molecular level,the complete nucleotide sequence of S6 was determined.The S6 comprised 2 030 nt and was predicted to encode a 68.4 kD protein.The result of cluster analysis revealed that the new isolate was a member of aquareovirus,but identity between it and representative strain GCRV873 was 33%.The deduced amino acid sequence shared highest identity（33%） with grass carp reovirus 873 strain,indicating that the new isolate was presumed to be a new type of GCRV.

关 键 词： 草鱼呼肠孤病毒 病毒分离 病毒鉴定

领　　域： [农业科学] [农业科学]