2009年深圳新甲型H1N1流感病毒分离鉴定及分离株特征分析  

Genetic Characterization of H1N1 Influenza A Virus Obtained from Shenzhen in 2009

作　　者：刘涛[1] 吴春利[1] 房师松[1] 黄运美[1] 吕星[1] 王昕[1] 邱亚群[1] 程小雯[1] 

机构地区：[1]深圳市疾病预防控制中心,广东深圳518020

出　　处：《实用预防医学》2010年第8期1528-1532,共5页Practical Preventive Medicine

摘　　要：目的比较新甲型H1N1流感病毒对MDCK细胞和鸡胚的敏感性差异,了解病毒分离株特征,为疫苗制备和开展实验室常规监测等奠定基础。方法用MDCK细胞和鸡胚进行流感病毒分离[1],收获病毒后提取病毒RNA,逆转录-聚合酶链式反应(RT-PCR)扩增,阳性扩增产物纯化后测序,测序结果使用Clustal和MEGA4.1软件进行比较分析,通过与NCBI数据库中不同时间和地区的流感病毒H1N1株同源比对后,绘制种系发生树。结果从334份患者呼吸道标本中,用MDCK细胞分离法分离出134株新甲型H1N1病毒,分离率为40.12%,用鸡胚分离法分离出150株新甲型H1N1病毒,分离率为44.91%;所测10株新甲型H1N1毒株HA1区的344个氨基酸,与2009年新甲型H1N1国际代表株A/California/07/2009相比,只有少数位点发生氨基酸替换,并且这些位点都不在抗原决定簇上;种系发生树显示,10株新甲型H1N1和国际代表株同源性较大,形成一簇,与A/Swine/guangxi/13/2006较为接近。结论新甲型H1N1病毒对MDCK细胞和鸡胚的敏感性相差不大;新甲型H1N1病毒未发生变异,短期内不会产生新变种。Objective To compare the susceptivity of novel H1N1 influenza virus on MDCK cell and chick embryos,and to identify the characteristics of influenza isolates for influenza vaccine developing.Methods The influenza virus was isolated from both MDCK cell and chick embryos,and reverse transcriptase polymerase chain reaction（RT-PCR）was conducted to identify the subtype of influenza virus.The sequences of novel H1N1 influenza virus were compared to NCBI database by using Clustal and MEGA4.1 software.Results Of 334 respiratory specimens,134 strains of novel H1N1 were isolated through MDCK cell culture,with the positive rate of 40.12%,and 150 strains were isolated through embryonated chicken＇s egg culture,with the positive rate of 44.91%.The sequences of 344 amino acids in 10 strains of novel H1N1 were obtained and then compared with international representative strain A/California/07/2009,only a few of sites were changed.After virus infection,it would take 9 days to produce protective antibodies.Conclusions The susceptivity of novel H1N1 influenza virus on MDCD cell and embryonated chicken＇s eggs is similar.Antigenic change is not found among the novel H1N1 strains.

关 键 词：新甲型H1N1流感 病毒分离 HA1基因 

分 类 号：R373.13[医药卫生—病原生物学]