牛病毒性腹泻病毒基因分型双重RT-PCR方法的建立  

Establishment of a Multiplex PCR Assay for Simultaneous Detection of BVDV Ⅰ and BVDV Ⅱ

作　　者：熊浩[1,2] 季新成[2] 王克栋[1] 史茜[2] 冉多良[2] 彭武丽[2] 于学辉[2] 

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [2]新疆出入境检验检疫局,新疆乌鲁木齐830063

出　　处：《中国动物检疫》2013年第4期39-42,共4页China Animal Health Inspection

基　　金：新疆自治区高新技术项目(项目编号201010101;201141147);国家质量监督检验检疫总局科研项目(项目编号2011IK017)

摘　　要：根据GenBank已发表的牛病毒性腹泻病毒(BVDV)基因Ⅰ型和Ⅱ型5’端非编码区(5’-UTR)保守序列,设计两对针对基因Ⅰ型和Ⅱ型的特异性引物,经反应条件的优化,建立了在同一反应体系中同时检测BVDV基因Ⅰ型和Ⅱ型的RT-PCR方法。该方法对两个基因型的病毒检测灵敏度均为2TCID50,只比单重PCR的灵敏度低了10倍。且具有较好的特异性和可重复性。经临床应用验证,该方法可用来对两个基因型的BVDV进行同步分型检测。Accoring to the conservative sequence of the 5＇-untranslated region （5＇-UTR） of BVDV genome in GenBank, two pairs of primers were designed for detecting BVDV Ⅰ and BVDV Ⅱ and an RT-PCR was developed for spontaneously detection of BVDV Ⅰ and Ⅱ in one reaction system after optimization of the reaction conditions.The results showed that the detection limit was 2 TCID50, only 10 times lower than the single PCR.Clinical application confirmed that the method had higher specifity and reliability and could be used to detect BVDV Ⅰ and BVDV Ⅱ rapidly and accurately.

关 键 词：牛病毒性腹泻病毒基因I型 牛病毒性腹泻病毒基因II型 双重RT-PCR 共扩增 

分 类 号：S852.653[农业科学—基础兽医学] S852.659.6[农业科学—兽医学]