利用巢式PCR快速鉴定牛传染性鼻气管炎

Fast Diagnosis of IBRV by Nested-PCR Method

为了快速检测牛传染性鼻气管炎,采用SDS-蛋白酶K法,提取病毒模板DNA。根据IBRV gB基因序列设计了1对特异引物,在其上下游引物的内侧又分别设计了1对引物,以这4条引物对IBRV模板进行扩增。结果成功扩增出预期目的片段,建立了巢式PCR检测方法。敏感性、特异性等检测试验结果表明该方法能特异检测IBR病毒。本方法具有快速、灵敏、特异的优点,适用于在牛及其遗传物质的进出口检疫中进行牛传染性鼻气管炎快速病原鉴定。

A nested-PCR method was established successfully to detect IBRV. Two pairs of primers were designed according to the published sequence of the Infectious Bovine Rhinotracheitis Virus （IBRV） gB gene. The external primer can amplify 478bp specific IBRV fragment and the internal primer can amplify 385bp fragments, which accords with the predicted amplification fragment. Moreover,it had no positive result with the system to amplify bovine viral diarrhea virus, bovine rotavirus, bovine vesicular stomatitis virus nucleotide, which demonstrate the PCR amplification system has good specificity. In addition,it also has very high sensitivity because the system can detect the 10-6 dilute template, manifesting latent application prospect.