污染疫苗的禽腺病毒分离和鉴定

Isolation and identification of avian adenovirus of contaminated vaccine

在用非免疫鸡胚增殖鸭病毒性肝炎病毒 (E5 2株 )过程中 ,发现种毒受到污染。采用聚乙二醇沉淀 SephadexG10 0柱层析法纯化污染病毒 ,在电镜下发现一大小为 70～ 80nm ,无囊膜 ,2 0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 99 5 %、 99 6 %和 99 5 % 。

During duck viral hepatitis seed virus(E52)was reproduced by non immunitied 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 amplify this contaminated virus by RT PCR method,three DNA fragments,showing 99 5%,99 6% and 99 5% 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 seed virus(E52)was contaminated by avian adenovirus.