BVDV荧光RT-PCR检测方法的建立及应用  

Establishment and application the fluorescence RT-PCR detection method of BVDV

作　　者：姚晓辉[1] 李华强[1] 陈瑞爱[1] 练炳洲[1] 王林川[1] 邓月嫦 王凤国[1] 

机构地区：[1]广东大华农动物保健品股份有限公司,广东新兴527400

出　　处：《广东农业科学》2011年第11期154-157,共4页Guangdong Agricultural Sciences

摘　　要：根据GenBank中登录的牛病毒性腹泻病毒(BVDV)毒株序列,选择5’NTR保守区域设计1对特异性引物和1条Taqman探针,通过矩阵法筛选引物探针的最佳浓度,建立了检测BVDV的荧光RT-PCR方法。结果显示,用1.5μL上下游引物混合液(10μmol/L)和0.5μL探针溶液(10μmol/L)对BVDV进行检测,可获得较小Ct值、较高吸光值,并可降低检测成本。对该方法进行特异性、敏感性和重复性试验,结果表明,该方法敏感性高、特异性强、重复性好。将该方法应用在疫苗生产中,可快速准确地筛选出合格的原辅材料和半成品,为生产提供及时准确可靠的数据。A pair of primers and a Taqman probe were designed based on the gene sequences of BVDV available in GenBank. The studies established the fluorescence RT-PCR detection methods of BVDV by screening the optimal concentration of the primers and probe. The results showed the method could obtain the smaller Ct value, high absorption value and reduce detection cost with 1.5 μL primers （10 μmol/L） and 0.5 μL probe （10 μmol/L）. And the experiments indicated the detection method had high specificity, strong sensitivity and good repeatability. The established method was applied the vaccine production, it could screen quickly the qualified raw materials and semi-finished products, and provide timely, accurate and reliable data for the production.

关 键 词：BVDV 荧光RT—PCR 检测方法 

分 类 号：Q789[生物学—分子生物学] S858.23[农业科学—临床兽医学]