禽流感、新城疫、传染性支气管炎病毒多联RT-PCR诊断方法的建立

利用分子生物学方法,选择禽流感病毒的M基因、新城疫病毒的F基因以及传染性支气管炎病毒的N基因,分别在基因序列的保守区域设计1对特异性引物,利用所合成的3对引物对相应靶基因进行RT-PCR扩增,在优化单项RT-PCR条件的基础上,建立禽流感、新城疫、传染性支气管炎病毒多联RT-PCR的检测方法,同时对该检测方法进行敏感性及特异性试验。结果表明:这3对引物分别能特异性地扩增出禽流感病毒的M基因片段、新城疫病毒的F基因片段及传染性支气管炎病毒的N基因片段,大小分别为385 bp、520 bp和748 bp,初步建立了快速、敏感、特异鉴别检测禽流感、新城疫、传染性支气管炎病毒的分子诊断方法。

Development of molecular method for differentiating and detecting AIV,NDV and IBV

To differentiate and detect avian influenza virus, newcastle disease and infectious bronchitis virus of the signifi- cant infectious disease caused by virus in avian rapidly, sensitively and distinctly. Based on their conserved regions of M gene of AIV, F gene of NDV and N gene of IBV registered in GenBank, three sets of specific primers were designed and synthesized. The results showed that three specific bands, about 385 bp, 520 bp and 748 bp individually, were amplified by multiplex RT- PCR of AIV,NDV and IBV with three sets of specific primers. By specificity and sensitivity test, the detection method was highly specific and sensitive. The molecular diagnose method was preliminarily established to detect AIV, NDV and IBV rapidly, sensitively and specifically.