一株锦鲤疱疹病毒的分离与鉴定  

Isolation and identification of Kio herpesvirus

作　　者：朱霞[1] 李新伟[1] 王好[1] 吕文亮[1] 周井祥[2] 

机构地区：[1]吉林农业大学动物生产及产品质量安全教育部重点实验室,吉林长春130118 [2]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中国预防兽医学报》2011年第5期340-343,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30671621)

摘　　要：本实验对患病鲤鱼进行病毒分离及鉴定,将表现典型锦锂疱疹病毒(KHV)病症状的鲤鱼肾细胞与单层框镜鲤鱼鳍细胞(KFC)共培养,5d～6d后出现典型的KHV细胞病变,即细胞体积增大并出现空泡化;在电镜下观察到典型的KHV病毒粒子;并应用PCR方法扩增获得KHVORF25基因的849bp基因片段,连接至pMD18-T载体中,经酶切鉴定后序列分析表明,与GenBank登录的3株KHV同源性均为99.9%,命名为KHV-cj。基因进化比较显示,与KHV-J株关系较近。将病毒培养物(10-6.75TCID50)腹腔注射接种实验鱼可使其100%发病,并且症状和病理变化与自然感染KHV的鱼一致,死亡率为75%。表明本研究在国内首次分离到KHV。A Koi herpesvirus （KHV） was isolated from carp suffering gill necrosis by co-cultivation of the affceted kidney cells with carp fin cells （CFC）. Cytopathic effects （CPE） were presented 5 days to 6 days post co-cultivation. The KHV particles were observed under electron microscope by negatively stained the sediment of virus culture with 0.5% phosphotungstate. A 849 bp DNA fragment of ORF25 gene was amplified by PCR and sequencing result indicated that the sequence of nucleotide acid homology was 99.9% with the reference strains of KHV in GenBank. The some clinic signs as nature infection were presented in fish artificially infected with 10-6.75 TCID50 by intraperitoneal injection and the morbility and mortality were 100% and 75%, respectively. This result demonstrated that the KHV was isolated for the first time from the naturally infected fish in China.

关 键 词：框镜鲤 锦鲤疱疹病毒 分离鉴定 

分 类 号：S852.65[农业科学—基础兽医学]