H5、H9亚型禽流感分子鉴别诊断方法的建立

Development of molecular method for differentiate detecting H5 and H9 subtype of Avian Influenza Virus

为了快速、敏感、特异地鉴别诊断禽流感,根据H5、H9亚型禽流感病毒HA基因序列的保守区域设计出并合成2对特异性引物,利用这2对引物对H5、H9亚型禽流感病毒的HA基因片段进行二联RT-PCR扩增,并对二联RT-PCR检测方法进行敏感性及特异性试验。结果表明:这2对引物能特异性地扩增出H5、H9亚型禽流感病毒HA基因片段,大小分别为490bp和686bp;该检测方法敏感性高,特异性强;初步建立了快速、敏感、特异地鉴别检测H5、H9亚型禽流感病毒的分子诊断方法。

Two pairs of specific primers were designed and synthesized according Io conserved regions of HA genes of H5 and H9 subtype of avian influenza viras registered in GenBank, The resnlts showed that,two specific bands,about 490 bp and 686 bp individually,were amplified by multiplex RT - PCR of H5 and H9 subtype of avian, influenza virus with the two sets of specific primers. By specificity and sensitivity test,the detection method was highly specific and sensitive. The molecular differentiation method was preliminarily established to detect H5 and H9 subtype of avian influenza virus rapidly, sensitively and specifically.