牛病毒性腹泻病毒、牛传染性支气管炎病毒和口蹄疫病毒多重TaqMan荧光定量PCR检测方法的建立

发布时间：2018-03-23 11:57

本文选题：牛病毒性腹泻病毒　切入点：牛传染性鼻气管炎病毒　出处：《中国预防兽医学报》2017年02期

【摘要】：为建立同时快速鉴别检测牛病毒性腹泻病毒(BVDV)、牛传染性支气管炎病毒(IBRV)和口蹄疫病毒(FMDV)的方法,本研究根据高度保守的BVDV的5'UTR基因、IBRV的g B基因和FMDV的3D基因,分别设计了3对对应的特异性引物和3种不同发光基团标记的Taq Man探针,建立了同时检测这3种病毒的多重荧光定量PCR的方法。并对其反应条件进行优化。结果表明,该多重荧光定量PCR方法能够特异性检测出BVDV、IBRV和FMDV,而对BTV等检测结果均为阴性,对BVDV、IBRV、FMDV的最低检测量分别为194拷贝/μL、208拷贝/μL和150拷贝/μL。而且组内和组间变异系数均低于0.85%。本研究所建立的多重荧光定量PCR具有方便、快速、特异性好、灵敏度高、重复性好等优点,能够用于BVDV、IBRV和FMDV的同时检测。
[Abstract]:In order to establish a method for rapid identification of bovine viral diarrhea virus (BV DVV), bovine infectious bronchitis virus (IBRV) and foot-and-mouth disease virus (FMDV) simultaneously, a highly conserved 5'UTR gene of BVDV, GB gene of IBRV, and 3D gene of FMDV, were used in this study. Three pairs of specific primers and three Taq Man probes labeled with different luminescence groups were designed, and a method for simultaneous detection of multiplex fluorescence quantitative PCR of these three viruses was established. The reaction conditions were optimized. The multiplex fluorescence quantitative PCR method can detect BV DVV IBRV and FMDV specifically, but the results of BTV and so on are negative. The lowest detection of IBRV FMDV was 194 copies / 渭 L, 208 copies / 渭 L and 150 copies / 渭 L. the coefficient of variation within and between groups was lower than 0.85. The multiplex fluorescence quantitative PCR established in this study was convenient, rapid, specific and sensitive. It can be used for simultaneous detection of BV DVV IBRV and FMDV.
【作者单位】：沈阳农业大学;中国动物疫病预防控制中心;
【分类号】：S852.65

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