作　　者： ; ; ; ; ; ; ;

机构地区： 中国水产科学研究院珠江水产研究所

出　　处： 《水生生物学报》 2011年第5期790-795,共6页

摘　　要： 从江西南昌患出血病草鱼体内分离出的草鱼病毒(暂命名为JX09-01)能使草鱼肾脏细胞(CIK)、草鱼肝细胞(L8824)、草鱼吻端成纤维细胞(PSF)产生明显的细胞病变效应(CPE)。感染CIK细胞固定后经电镜观察,发现细胞质内有大量病毒聚集,形态和排列方式与已报道的草鱼呼肠孤病毒(Grass carp reovirus,GCRV)相似。针对GCRV 873株S6基因设计的简并引物可以从病料组织和感染细胞中扩增出目的条带,而针对GCRV HZ08株S6基因设计特异性引物未能扩增出目的条带。对JX09-01株的S6全基因进行序列分析表明,其核苷酸序列同GCRV 873株和HZ08株的同源性分别是99.3%和30.4%,推导出的氨基酸序列同源性分别是98.6%和30%,说明草鱼病毒JX09-01株为草鱼呼肠孤病毒。用JX09-01株接种当年8—10 cm左右的草鱼,没有明显的临床症状,不能致草鱼死亡。用传代至15代的CIK细胞病毒液进行免疫保护试验,结果显示其对强毒株的免疫保护率达到86.7%。实验结果初步显示,新分离到的JX09-01为草鱼呼肠孤病毒弱毒株,可作为弱毒疫苗的候选毒株。 A new grass carp virus,assigned JX09-01,was isolated from a diseased grass carp case during routine ex-amination in Nanchang City,Jiangxi Province.The obvious cytopathic effects（CPE） were observed after the new iso-late inoculated in grass carp kidney cell（CIK）,grass carp liver cell（L8824） and proboscis fibroblast cell（PSF） respec-tively,and plenty of non-enveloped virus particles with hexagonal shape were observed under electron microscope（EM）,which was highly similar to reported grass carp reovirus（GCRV）.The specific bands could be detected by reverse transcription-polymerase chain reaction（RT-PCR） from infected fish and cells using primer designed according to the sequences of GCRV 873 strain,but no result obtained using primers designed with reference to the sequence of GCRV HZ08 strain.Sequence analysis of segment 6 showed that the nucleotide sequence similarity between JX09-01 and other GCRV strains was 99.3%（GCRV 873） and 30.4%（GCRV HZ08）,while the deduced amino acid sequence similarity were 98.6% and 30% respectively,which indicated the JX09-01 is grass carp reovirus.The pathogenicity test on grass carp found that new separated virus was a mild strain.The grass carps with 8—12 cm length were immunized with the new isolate and protection rate of 86.7% was obtained against virulent strain challenge,which indicated that the new separated strain could be a candidate for attenuate vaccine strain as a grass carp reovirus.

关 键 词： 草鱼呼肠孤病毒 分离 鉴定 序列分析 免疫原性

领　　域： [农业科学] [农业科学]