禽流感病毒油乳剂灭活疫苗的核酸抽提和荧光RT-PCR检测  

Nucleic Acid Extraction and Fluorescence RT-PCR Detection of Avian Influenza Virus Oil Emulsion Inactivated Vaccine

作　　者：段倩倩 王军[1] 周迎春[1] 何长生[1] 朱良强[1] DUAN Qianqian;WANG Jun;ZHOU Yingchun;HE Changsheng;ZHU Liangqiang(Anhui Provincial Center for Animal Disease Control and Prevention,Hefei Anhui 230091,China)

机构地区：[1]安徽省动物疫病预防与控制中心,安徽合肥230091

出　　处：《畜牧兽医杂志》2023年第6期6-8,共3页Journal of Animal Science and Veterinary Medicine

基　　金：安徽省动物疫情监测与防治项目。

摘　　要：本研究旨在研究禽流感病毒(Avian Influenza Virus,AIV)灭活疫苗中是否存在可被检测到的核酸成分,分析其污染环境的可能性,以重组AIV油乳剂灭活疫苗为研究对象,经离心法、反复冻融法分离水相,倍比稀释后抽提核酸,采用荧光RT-PCR测定AIV核酸阳性的最高稀释度。结果显示:反复冻融法分离AIV灭活疫苗水相效果较离心法好,将疫苗水相稀释4\+12倍后仍能检测到AIV核酸阳性。结果表明:微量AIV灭活疫苗中的核酸仍可被荧光RT-PCR检出,疫苗污染可以作为环境样品“假阳性”的排查因素之一。The purpose of this study is to investigate whether there are detectable nucleic acid components in the inactivated vaccine of avian influenza virus(Avian Influenza Virus,AIV),and to analyze the possibility of its contamination of the environment.In this study,the recombinant AIV oil emulsion inactivated vaccine was used as the research object.The aqueous phase was separated by centrifugation and repeated freezing and thawing,and the nucleic acid was extracted after doubling dilution.Finally,fluorescent RT-PCR was used to determine the highest dilution of AIV nucleic acid positive.The results showed that the repeated freeze-thaw method was better than the centrifugation method for separating the aqueous phase of AIV inactivated vaccine.We can still detect AIV nucleic acid positive after diluting the vaccine aqueous phase by 4^(12) times.The results showed that the nucleic acid in trace amounts of AIV inactivated vaccine could still be detected by fluorescent RT-PCR,and vaccine contamination could be used as one of the screening factors for"false positive"in environmental samples.

关 键 词：禽流感病毒 灭活疫苗 疫苗污染 核酸检测 假阳性 

分 类 号：S852.657[农业科学—基础兽医学]