牛病毒性腹泻病毒一步法RT-PCR检测方法的建立与应用

为建立一种快速检测牛病毒性腹泻病毒病原的方法,本研究根据GenBank上登录的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)基因组序列,设计1对引物,建立了检测BVDV的一步法RT-PCR方法。该方法对牛传染性鼻气管炎病毒、猪瘟病毒、牛副流感病毒3型的扩增结果均为阴性,检测的敏感性达1 ng RNA。该一步法RT-PCR方法具有良好的特异性、敏感性、重复性,可以准确快速检测出极低含量的BVDV,将为BVDV的病原检测及分子流行病学调查等提供一种快速、灵敏、特异、准确的分子生物学检测方法。

Development and Application of One Step RT-PCR Method for Detection of Bovine Viral Diarrhea Virus

To establish a rapid bovine viral diarrhea virus (BVDV) pathogen detection methods, based on BVDV gene sequence in GenBank, we synthesized one pair of primers, established one step RT-PCR for BVDV detection. The method was used for infectious bovine rhinotracheitis virus (IBRV), classical swine fever virus(CSFV), bovine para influenza virus 3 (BPIV3), all of the PCR results were negative, the sensitivity was 1 ng RNA. The established one step PCR method had good specificity, sensitivity, reproducibility, it could detect very low levels of BVDV quickly and accurately, it provided a kind of rapid, sensitive, specific and precise molecular biology detection method for pathogen detection and molecular epidemiology of material such as BVDV.