H5、H7和H9亚型禽流感分子鉴别诊断方法的建立

Establishment of Molecular Method for Differentiating and Detecting H5、H7and H9 Subtype of Avian Influenza Virus

为快速、敏感、特异地鉴别诊断现较为重要的禽流感病毒亚型,根据H5、H7和H9亚型禽流感病毒HA基因序列的保守区域设计出并合成3对特异性引物,利用这3对引物对H5、H7和H9亚型禽流感病毒的HA基因片断进行多联RT-PCR扩增,并对多联RT-PCR检测方法进行敏感性及特异性实验。结果表明:3对引物能特异性地扩增出H5、H7和H9亚型禽流感病毒HA基因片段,大小分别为490bp,365bp和686bp;该检测方法敏感性高,特异性强;初步建立起快速、敏感、特异地鉴别检测H5、H7和H9亚型禽流感病毒的分子诊断方法。

To differentiate and detect the significant subtype of avian influenza virus rapidly, sensitively and distinctly, Based on their conserved regions of HA genes of H5. H7and H9 subtype of avian influenza virus registered in GenBank,three sets of specific primers were designed and synthesized. The results showed thatthree specific bands, about 490 bp ,365 bp and 686 bp respectively, were amplified by multiplex RT-PCR of H5,H7 and H9 subtype of avian influenza virus 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 H5,H7 and H9 subtype of avian influenza virus rapidly,sensitively and specifically.