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目的了解2013年深圳市新甲型H1N1流感病毒的分子变异情况。方法用MDCK细胞进行流感病毒分离,收获病毒后提取病毒RNA作为模板,利用RT-PCR对其HA基因进行扩增,扩增产物经琼脂糖凝胶电泳鉴定后测序,测序结果用Clustal W2和MEGA3.1软件进行序列分析。结果 2013年的深圳毒株与同年国内外的流行株差异不大,但与同期国际疫苗株A/California/7/2009相比,在进化树上已经产生了明显的分离,且出现了2个不同的进化分支;HA分子有多个氨基酸位点发生了变异,其中有4个变异位点位于抗原决定簇区域内:抗原决定簇Sa的135位点由N变成了S,163位点的K出现了N/Q/I的多元突变;抗原决定簇Sb的185位点由S变成了T;抗原决定簇Ca1的203位点由S变成了T,这些变异的出现会导致病毒抗原性的改变。结论 2013年深圳市流行的新H1N1病毒有较大可能会形成一个新的优势流行株。
Objective To understand the molecular variation of new influenza A (H1N1) virus in Shenzhen in 2013. Methods The virus was isolated from MDCK cells and the virus RNA was harvested. The viral RNA was extracted and used as a template. The HA gene was amplified by RT-PCR. The amplified product was identified by agarose gel electrophoresis and sequenced. The sequencing results were performed with Clustal W2 and MEGA3 .1 software for sequence analysis. Results The Shenzhen strain in 2013 was not significantly different from the epidemic strains at home and abroad in the same year. However, compared with the international vaccine strain A / California / 7/2009 in the same period, there was a clear separation in the evolutionary tree and two Different evolutionary branches. There are several amino acid sites in HA molecule that have been mutated. Among them, four mutated sites are located within the antigenic determinant region: the 135 site of antigenic Sa changed from N to S, K showed a multiple mutation of N / Q / I; epitope 185 of Sb changed from S to T; the site 203 of Ca1 of the antigenic cluster changed from S to T, and the occurrence of these mutations would lead to viral antigen Sexual changes. Conclusion The new H1N1 virus prevailing in Shenzhen in 2013 is likely to form a new prevalent epidemic strain.