甲型H1N1流行性感冒病毒血凝素基因变异状况及序列分析  

Haemagglutinin gene mutations and sequences of influenza viruses A （H1N1）

作　　者：侯佩强[1] 张荣强[1] 边广[1] 陆娟娟[1] 

机构地区：[1]山东省泰安市疾病预防控制中心,271000

出　　处：《中华传染病杂志》2011年第11期679-684,共6页Chinese Journal of Infectious Diseases

基　　金：泰安市科学技术发展计划项目（20071034）

摘　　要：目的了解2009年度甲型H1N1流行性感冒（流感）病毒的检测情况和血凝素（HA）基因变异情况。方法选择国家级流感监测哨点医院以及暴发疫情的疫点，采集流感样病例的鼻咽拭子标本，通过实时（RT）-PCR进行病毒分型及甲型H1N1流感病毒检测，对阳性标本采用狗肾细胞（MDCK）进行病原分离，采用红细胞凝集试验测定病毒效价，用血凝抑制实验进行型别鉴定，通过RT—PCR扩增毒株HA1片段的基因并进行序列测定，利用生物信息学技术进行序列分析。结果共检测咽拭子样本996份，其中核酸检测阳性病例包括甲型H1N1337份，季节性H1N1亚型1份，季节性H3N2亚型67份，B型12份，流感核酸检测阳性率为41．87％，其中甲型流感核酸检测阳性率为33．84％。分离出甲型H1N1病毒36株，选择18株。测序成功的10株甲型H1N1流感病毒在多个氨基酸位点发生变异，与疫苗株A／California／07／2009（H1N1）比较，有6个位点发生突变，其中1个位点位于抗原决定簇的B区。结论2009年度分离到的流感病毒株中以甲型H1N1为绝对的优势毒株，毒株的血凝素基因与世界卫生组织（WHO）提供的疫苗株相比有变异，与疫苗株相比，抗原决定簇B区有改变，但关键位点第222位没有变化。Objective To understand the detections of influenza A （H1N1） in 2009, and haemagglutinin （HA） gene mutations and the comparisons with standard strains. Methods The nasopharyngeal swabs from patients with influenza-like illness （ILI） in National Influenza Sentinel Surveillance Hospital and the outbreak epidemic area were collected. The virus typing and A （H1N1） viruses were tested by real time-polymerase chain reaction （RT-PCR）. Then the pathogens were isolated with MDCK cells, the virus titer was determined with hemagglutination test and the virus typing was identified with hemagglutination inhibition test （HAI）. The RT-PCR products of HA1 gene of virulent strains were sequenced and then analyzed through bioinformatics. Results A total of 996 pharyngeal swab specimens were tested, and nucleic acid positive cases included 337 A （H1N1） subtype, 1 seasonal A （HIN1） subtype, 67 A （H3N2） subtype, and 12 B type. The positive rate of nucleic acid detection of influenza was 41. 87% and that of A （HIN1） was 33. 84%. Thirty-six influenza A （H1N1） virus strains were isolated, and 10 of them were successfully sequenced and several amino acid mutations were identified. There were 6 amino acid mutations found compared with vaccine strain A/California/07/2009 （H1N1）, and 1 site was in area B of epitope. Conclusions A （H1N1） is absolute predominant among isolated strains in 2009. HA gene of virulent strains is rout area ated compared with vaccine strain provided by World Health Organization, which shows that the B of epitope changes, while the key amino acid position 222 doesn＇t change.

关 键 词：流感病毒A型 H1N1亚型 基因 病毒 血凝素类 病毒 聚合酶链反应 序列分析 变异(遗传学) 

分 类 号：R51[医药卫生—内科学]