鲤科疱疹病毒Ⅱ型射阳株的分离与鉴定  

Isolation and identification of a strain of Cyprinid Herpesvirus 2 in Sheyang

作　　者：李茂 肖丹 刘天强 刘衍鹏 黄冠军 汪开毓[2] 罗远会 

机构地区：[1]通威股份动物保健研究所,成都610041 [2]四川农业大学鱼病研究中心,四川雅安625014

出　　处：《淡水渔业》2015年第3期93-96,共4页Freshwater Fisheries

基　　金：通威股份有限公司科研专项资金(TW2012E001)

摘　　要：对患＂鳃出血＂病的异育银鲫（Carassius auratus gibelio）病灶组织进行分离,并通过回归感染试验、细胞分离鉴定、电镜观察以及PCR几种方法结合来鉴定。结果显示：健康异育银鲫注射组织滤液5 d后开始发病,且症状与原发病症状一样,死亡率为80%,对照组没有死亡。对患病鱼内脏组织经超薄切片,电子显微镜观察发现组织内有大量的病毒颗粒,成熟的病毒粒子直径大小170~200 nm,与疱疹病毒Ⅱ型病毒粒子相符。滤液经疱疹病毒Ⅱ型特异性的PCR检测,获得阳性目的片段,同时将无菌处理的病样滤液接种胖头鲤细胞（FHM）,培养1 d后出现明显的细胞病变（CPE）,单层细胞网状收缩,坏死细胞聚集。收集细胞培养液,进行病毒特异性的PCR检测,并检测到436 bp的阳性片段。结果表明：从江苏射阳某＂鳃出血＂病异育银鲫养殖鱼塘分离到的病毒为疱疹病毒Ⅱ型（Cy HV-2）病毒。In order to verify that the isolated virus was CyHV-2 virus, the test of the gill hemorrhage disease of Carassius auratus gibelio lesions were isolated and identified by recurrent infection test , isolation and identification , electron microscope observation and PCR for identification of herpes simplex Ⅱtype virus.The results showed： Health C.auratus gibelio injection of tissue filtrates of 5 d after the beginning of the disease , and symptoms of the primary disease , the mortality rate was 80%, the control group had no death .The prevalence of fish viscera tissues by ultrathin sections within the organization, found that a large number of virus particles observed by electronic microscope , mature virus particle diameters of 170-200 nm, and herpes simplex Ⅱtype virus particle match .Detection of the filtrate by herpes simplex Ⅱtype virus specific polymerase chain reaction （PCR）, and detected the positive fragment of 436bp, at the same time the filtrate inoculation fat carp cell disease like aseptic processing （ FHM） , after 1 d culture appears obvious cytopathic （ CPE） , cell monolayer mesh shrinkage , necrosis cell aggregation .The collection of cell culture fluid , detection of virus specific polymerase chain reaction （PCR）, and detected the positive fragment of 436bp.Conclusion： All of the evidences indicated that the isolated virus stain was CyHV-2.

关 键 词：鲤科疱疹病毒Ⅱ型 异育银鲫(Carassius auratus gibelio) 分离 鉴定 

分 类 号：S917.1[农业科学—水产科学]