云南省边境地区禽流感H5N1亚型病毒基质蛋白2基因进化分析  

Genetic evolution analysis of matrix protein 2 gene of avian influenza H5N1 viruses from boundary of Yunnan province

作　　者：肖雪[1,2] 张文东[2] 段博芳[2] 赵焕云[2] 刘庆亮[1,2] 胡挺松[3] 邱薇[3] 冯子良[3] 郑颖[3] 范泉水[3] 张应国[2] 张富强[3] 

机构地区：[1]云南农业大学,昆明650201 [2]云南省动物疫病预防控制中心 [3]昆明市成都军区疾病预防控制中心

出　　处：《中华预防医学杂志》2013年第6期514-517,共4页Chinese Journal of Preventive Medicine

基　　金：国家公益性行业专项（201103008）;军队科技计划项目（A12005,CWS12J075,AWS11L009）;云南省科技计划项目（2012CH002）

摘　　要：目的了解2008--2012年云南省边境地区禽流感H5N1亚型病毒基质蛋白2（M2）和离子通道蛋白基因变异特征及遗传进化关系。方法于2008--2012年采集云南省边境地区（越南、老挝、缅甸与云南省交界处）家禽和野生鸟类棉拭子样品1240份，经H5N1亚型特异性多重RT—PCR检测，对阳性样品病毒M基因进行扩增，克隆至pMD18-T载体测序，获得M2基因序列，并与已知参考毒株序列进行序列比对及系统发育分析。结果自1240份样品中检出H5N1亚型阳性样品71份，阳性率为5．72％；选择其中的30份阳性样品进行病毒M2基因测序，共获得14种不同序列，存在3个不同进化分支：1．2．1、1．2．2、2，各有5、7、2份。通过基因系统发育分析发现，M2基因与HA基因呈现不同进化关系；M2蛋白涉及表位、金刚烷胺抗性和禽源、人源毒株的关键性氨基酸位点存在替代或突变。结论2008--2012年期间云南边境H5N1亚型病毒M2基因具有遗传差异，M2基因进化分支1．2．2毒株已成为当地流行的优势毒株。Objective To elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein（M2） genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012. Methods A total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos, myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT- PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains. Results A total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained fronl 30 positive samples and were divided into 3 distinct clades or sub-clades （ 1.2. 1, 1.2.2 and 2） by phylogenetic analysis,5,7 and 2,respectively. The M2 genes and Hemagglutinin（HA） genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains. Conclusion The M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2. 2 had become dominant epidemic strain in this region.

关 键 词：正粘病毒科 流感病毒A型 H5NI亚型 病毒基质蛋白质类 遗传分析 

分 类 号：S852[农业科学—基础兽医学]