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目的研究杭州地区2009年至2012年甲型H1N1流感病毒血凝素(HA)与神经氨酸酶(NA)的基因进化特征,分析该病毒的遗传变异和抗原的分子水平转变。方法用RT-PCR分别扩增甲型H1N1流感病毒HA基因和NA基因片段并进行测序,用DNAMAN和MEGA 4.0生物信息学软件对HA和NA基因序列进行进化分析。结果建立的RT-PCR可成功检测HA(C)、HA(D)和NA基因,遗传进化分析结果表明,杭州地区甲型H1N1流感病毒的HA与NA氨基酸序列的亲缘系数与WHO推荐的病毒株及国内代表株的同源性均为98.9%～100%;三维构象结果表明,HA编码的蛋白质有5个位点发生改变,而NA编码的蛋白质仅发生2个位点改变。结论 2009年至2011年杭州地区甲型H1N1流感病毒HA和NA基因序列与世界范围内流行的病毒参考株具有高度同源性,但HA与NA蛋白质表位存在某些氨基酸位点的改变。
Objective To study the gene evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) of influenza A (H1N1) virus in Hangzhou from 2009 to 2012 and analyze the genetic variation of the virus and the molecular level transition of the antigen. Methods The HA gene and NA gene of Influenza A (H1N1) virus were amplified by RT-PCR and sequenced. The DNA and MEGA 4.0 bioinformatics software were used to analyze the HA and NA gene sequences. Results The results of RT-PCR showed that HA (C), HA (D) and NA genes were successfully detected. The results of genetic evolution analysis showed that the genetic relationship between HA and NA amino acid sequence of H1N1 influenza virus in Hangzhou was similar to that of WHO recommended virus And the homology of the representative strains in China were 98.9% -100%. The three-dimensional conformational results showed that there were 5 sites in HA-encoded protein, while only 2 sites in NA-encoded protein. Conclusion The sequences of HA and NA genes of influenza A (H1N1) virus in Hangzhou area from 2009 to 2011 are highly homologous to the reference strains of viruses that are popular in the world. However, there are some amino acid changes in HA and NA protein epitopes.