快速检测牛病毒性腹泻病毒实时荧光定量PCR技术建立及应用  

The Establishment and Application of Real-time Fluorescent Quantitative PCR Method to Rapidly Detect Bovine Viral Diarrhea Virus

作　　者：郭锐[1] 陈平 田永祥[1] 杨克礼[1] 刘泽文[1] 段正赢[1] 梁望旺[1] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室,武汉430064 [2]湖北出入境检验检疫局,武汉430050

出　　处：《湖北农业科学》2011年第8期1640-1643,共4页Hubei Agricultural Sciences

基　　金：湖北省农业科技创新中心资助项目(2007-620-001-03);动物胚胎工程及分子育种湖北省重点实验室开放课题项目(2010ZD158)

摘　　要：以牛病毒性腹泻病毒(BVDV)的保守基因序列为参考,设计、优化出一对特异的PCR引物和一条TaqMan荧光探针,建立一种快速定量检测牛病毒性腹泻病毒的实时荧光定量PCR技术。该方法检测灵敏度比RT-PCR高100倍,并且避免了常规PCR电泳检测所带来的高污染率。因此,该方法具有快速、灵敏、特异、重复性好和能定量检测等优点,适用于牛场BVDV感染的快速定量检测和肉类食品进出口检疫。A fluorescent quantitative PCR（FQ-PCR） method based on sequences of the BVDV genome was established to rapidly detect the bovine viral diarrhea virus.It included the disignation and optimization of a pair of specific primers and a fluorescent TaqMan probe for convseved gene.Comparation test showed that the sensitivity of this method was 100 times higher than RT-PCR test;and it could decrease the contamination usually caused by other conventional PCR.In conclusion,the FQ-PCR method was rapid,sensitive,specific and accurate;and thus could be used for rapidly detection of BVDV from cattle＇s tissues and other meat products.

关 键 词：牛病毒性腹泻病毒 实时荧光定量PCR 定量检测 

分 类 号：S858.23[农业科学—临床兽医学]