pdf原文污染疫苗的禽腺病毒分离和鉴定.pdfVIP

南京农业大学学报 　2004 , 27 (3) : 78～80 　 Journal of N anjin g A gricultural U niversity 污染疫苗的禽腺病毒分离和鉴定 周斌, 刘华雷, 曹瑞兵, 陈溥言 (南京农业大学农业部动物疫病诊断与免疫重点开放实验室 , 江苏 南京 210095) 摘要 : 在用非免疫鸡胚增殖鸭病毒性肝炎病毒 ( E52 株) 过程中 , 发现种毒受到污染。采用聚乙二醇沉淀Sephadex G100 柱层析法纯化污染病毒 , 在电镜下发现一大小为 70 ～80 nm , 无囊膜 , 20 面体对称的病毒颗粒。挑选 4 条随机引 ( ) 物对其进行反转录 PCR 扩增 , 共扩增出 3 条基因片段。序列分析结果表明 , 与禽腺病毒 鸡胚致死孤儿病毒 基因组部 分序列同源性分别高达 995 % 、996 %和 995 % , 从而证明禽腺病毒污染了鸭病毒性肝炎病毒鸡胚尿囊液种毒。 关键词 : 禽腺病毒 ; 分离 ; 纯化 ; PCR ; 鉴定 中图分类号: S85265 + 91 　　　文献标识码 : A 　　　文章编号 : 1000 2030 (2004) 03 0078 03 Isolation and identification of avian adenovirus of contaminated vaccine ZHOU Bin , L IU Hualei , CAO Ruibing , CHEN Puyan ( Key Laboratory of Animal Disease Diagnostic and Immunology , Ministry of Agriculture , Nanjin g Agricultural University , Nanjin g 210095 , China) Abstract : During duck viral hepatitis seed virus ( E52) was reproduced by nonimmunitied embryo , a contaminated virus , which could infect chicken was detected. After purification by PEG6000 , Sephadex G100 gel chromatography and dialysis , symmetry nonenveloped icosahedral virons with a diameter of 70 to 80 nm were found. Four random primers were selected to amplif y this contaminated virus by RTPCR method , three DNA fragments , showing 995 % , 996 % and 995 % homologue respectively with the genome of avian adenovirus (chicken embryo orphan lethal virus) were got by the sequencing analysis. These results suggested that the used duck viral hepatitis