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目的建立并完善贵州省新型甲型H1N1流感病毒基因分析检测技术平台,了解2009～2010年度贵州省新型H1N1流感病毒HA1片段与WHO公布的流行株是否有发生变异。方法收集10株新型H1N1流感病毒毒株,提取病毒核酸RNA,通过RT-PCR扩增HA1片断,用电泳观察PCR产物目的条带的大小,经过纯化、测序,用Biodeit、MEGA4相关软件进行序列比对,同源性及差异性分析,构建系统发育树。结果与古典猪流感病毒株相比,贵州省2009年、2010年、2011年流感病毒株同源性分别为72.0%～74.6%、72.0%、67.8%～68.3%。与其他4株亚洲株H1N1流感病毒代表株相比,贵州省2009年、2010年、2011年流感病毒株同源性分别为92.3%～95.8%、91.5%～92.4%、89.8%～90.8%。2009年度组内同源性为93.2%～100%;2010年组内高度同源,为100%;2011年组内同源性为98.3%。贵州省流感毒株与中俄代表株A/Habarovak/01/2009(H1N1)的HA1基因同属一个谱系,在同一个分支上,同时,与墨西哥第一株甲型H1N1流感A/MexioCity/001/2009(H1N1)亲缘关系也较近,在同一个大分支上。结论贵州省新型H1N1流感病毒代表株HA1基因与WHO公布的流行株在核苷酸水平有一定持续水平的变异。
Objective To establish and perfect a new platform for gene analysis and detection of Influenza A (H1N1) virus in Guizhou Province, and to find out whether there is any mutation in the HA1 fragment of the novel H1N1 influenza virus in Guizhou Province and that of the WHO published in 2009 and 2010. Methods Ten novel strains of H1N1 influenza virus were collected and the RNA of viral RNA was extracted. The HA1 fragment was amplified by RT-PCR. The size of PCR products was observed by electrophoresis. After purification and sequencing, the sequences were compared with Biodeit and MEGA4 software Homology, homology and diversity analysis, the construction of phylogenetic tree. Results Compared with the classical swine influenza virus strains, the homology of influenza virus strains in Guizhou Province in 2009, 2010 and 2011 were respectively 72.0% -74.6%, 72.0% and 67.8% -68.3%. Compared with the representative strains of four other Asian strains of H1N1 influenza virus, the homology of influenza virus strains in Guizhou Province in 2009, 2010 and 2011 were 92.3% -95.8%, 91.5% -92.4% and 89.8% -90.8%, respectively. In 2009, the homology in the group was 93.2% -100%. In 2010, the homology was 100% in the group. In 2011, the homology in the group was 98.3%. The HA1 gene of Guizhou province was associated with the HA1 gene of A / Habarovak / 01/2009 (H1N1) of China and Russia on the same branch. At the same time, the HA1 gene was linked to the first A / MexioCity / 001 / 2009 (H1N1) kinship is also close, on the same big branch. Conclusion The HA1 gene of the novel H1N1 influenza virus in Guizhou Province has a consistent level of nucleotide variation with the published WHO epidemic strains.