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

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.