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.
出处
《中国动物检疫》
CAS
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)