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目的研究邯郸市2012年流感监测中分离的Yamagata系乙型流感毒株HA1的抗原性和基因特性,阐明HA1基因的变异与流感流行的关系。方法用MDCK细胞分离培养流感病毒,提取病毒核酸,采用RT-PCR方法特异性扩增病毒HA1基因,进行核苷酸序列测定,用DNAStar中Megalign软件进行分析。结果2012年分离的Yamagata系乙型流感病毒与2008~2009年代表株B/Florida/4/2006相比亲缘关系较远,核苷酸同源性为96.2%~96.8%,氨基酸同源性为96.5%~97.4%,HA1区9个位点氨基酸发生替换,其中6个参与抗原表位构成;与WHO推荐的2012~2013年疫苗株B/Wisconsin/01/2010相比亲缘关系近,核苷酸同源性为98.8%~99.4%,氨基酸同源性为97.4%~98.5%,有5个位点发生氨基酸替换,其中3个参与抗原表位的构成;与国家流感中心提供的标准抗血清的血凝抑制滴度≥1︰640。结论邯郸市2012年分离的Yamagata系乙型流感病毒血凝素蛋白HA1与B/Florida/4/2006相比已形成新的变种,与B/Wisconsin/01/2010相比也发生了新的变异,但尚不能确定是否形成Yamagata系有代表性的新变种。
Objective To study the antigenicity and genetic characteristics of HA1 isolated from influenza virus strain Yamagata in 2012 in Handan City and to elucidate the relationship between HA1 gene mutation and the prevalence of influenza. Methods The influenza virus was isolated and cultured with MDCK cells. The viral nucleic acid was extracted. The viral HA1 gene was amplified by RT-PCR. The nucleotide sequence was determined by DNAStar Megalign software. Results The Yamagata influenza B virus isolated in 2012 was distantly related to the B / Florida / 4/2006 strain from 2008 to 2009. The nucleotide homology was 96.2% ~ 96.8%. The amino acid homology was 96.5% -97.4%. Amino acid substitution occurred at 9 sites in HA1 region, of which 6 were involved in antigenic epitope formation. Compared with the WHO-recommended vaccine strain B / Wisconsin / 01/2010 from 2012 to 2013, Acid homology was 98.8% ~ 99.4%, amino acid homology was 97.4% ~ 98.5%, amino acid substitution occurred in 5 sites, of which 3 involved in the composition of the epitope; with the National Influenza Center standard antiserum Hemagglutination inhibition titer ≥ 1: 640. Conclusions A new variant of Yamagata influenza virus hemagglutinin HA1 isolated from B / Florida / 4/2006 was found in Handan in 2012, and a new mutation was observed in comparison with B / Wisconsin / 01/2010 , But it is not yet certain whether or not to form a representative new variant of Yamagata.