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目的分离甲型H1N1流感病毒,分析金华市首例甲型H1N1流感死亡病例病毒分离株HA基因序列特征,为研究病毒进化、致病性、流行规律提供科学依据。方法采用MDCK细胞和real-time PCR法进行病毒分离、鉴定;提取病毒RNA,通过RT-PCR扩增其HA基因片段,测定核苷酸序列;利用GeneBank中相关序列对病毒分离株A/Jinhua/01/2009(H1N1)进行基因进化树分析。结果从3份咽拭子样本中分离出1株甲型H1N1流感病毒。HA基因序列分析证明:该毒株与2009年大流行株高度同源,与古典型猪流感亲缘性较近,与欧亚地区分离的A/H1N1猪流感病毒及A/H1N1禽流感病毒进化关系较远。结论金华市首例甲型H1N1流感死亡病例分离病毒株与北美流行株高度同源,可能是由古典型猪流感进化而来,与WHO选定的甲型H1N1疫苗候选株同源性较高,对于人季节性流感A/H1N1疫苗可能不敏感。
OBJECTIVE: To isolate the influenza A (H1N1) virus and analyze the sequence of HA gene in the first case of death from influenza A (H1N1) virus in Jinhua City, providing a scientific basis for studying the evolution, pathogenicity and prevalence of the virus. Methods The virus was isolated and identified by MDCK cells and real-time PCR. The viral RNA was extracted and the HA gene fragment was amplified by RT-PCR. The nucleotide sequence of the HA gene was determined. 01/2009 (H1N1) for phylogenetic tree analysis. Results One influenza A (H1N1) virus was isolated from three throat swab samples. HA gene sequence analysis showed that the strain was highly homologous to the 2009 pandemic strain and close to the classical swine influenza and was closely related to the evolution of the A / H1N1 swine influenza virus and the A / H1N1 bird flu virus in the Eurasian region Far. Conclusions The first case of death from type A H1N1 influenza in Jinhua City is highly homologous with the North American epidemic strain. It may have evolved from the classical swine flu and has high homology with the WHO candidate H1N1 vaccine candidate strain. May be insensitive to human seasonal influenza A / H1N1 vaccine.