云南边境禽流感H5N1亚型病毒血凝素基因进化特征分析  

Evolutionary characteristics of hemagglutinin genes of H5N1 subtype avian influenza viruses from boundary region of Yunnan Province

作　　者：叶丛华[1] 张文东[2] 宋建领[3] 李静[1] 田建国[1] 吕粤[2] 岳亮[2] 邱薇[4] 李刚山[4] 冯子良[4] 郑颖[4] 张思东[2] 范泉水[4] 张应国[2] 王双印[4] 张富强[4] 

机构地区：[1]云南农业大学,昆明650223 [2]云南省动物疫病预防控制中心,昆明650051 [3]云南省热带亚热带动物病毒病重点实验室,昆明650224 [4]成都军区疾病预防控制中心,昆明650032

出　　处：《中国卫生检验杂志》2009年第6期1216-1218,1247,共4页Chinese Journal of Health Laboratory Technology

基　　金：全军科技攻关项目(08G034);云南省科技攻关项目(2006NG-24);云南省后备人才基金项目(2005PY01-12)

摘　　要：目的：认识2003年～2008年期间云南边境禽流感H5N1亚型病毒血凝素（HA）基因的进化特征。方法：2003年～2008年期间在云南边境采集境外禽类样品380份,经H5/N1亚型特异性多重RT-PCR检测,对阳性样品中H5N1病毒HA基因进行RT-PCR扩增,克隆至pMD18-T载体测序,并与国内外已知参考毒株序列进行比对及系统发育分析。结果：多重RT-PCR检出18份阳性样品,阳性率4.74%;14份代表性病毒样品HA基因核苷酸和氨基酸序列同源性介于93.1%～99.3%、90.5%～99.1%;HA裂解位点序列存在4种不同排列方式,均具有高致病性禽流感病毒分子结构特征;系统发育分析表明阳性样品可划分为5个不同进化分枝（1、2.4、2.3.2、2.3.4、7）。结论：2003年～2008年期间云南边境境外H5N1亚型病毒间具有遗传差异,进化分枝2.3.4毒株已成为当地流行的优势毒株。Objective ：To elucidate evolutionary characteristics of the hemagglutinin （HA） genes of HSN1 subtype avian influenza viruses from boundary region of Yunnan province. Methods ：380 samples collected from foreign poultry in boundary region of Yunnan province during 2003 to 2008 were screened by H5/N1 subtype - specific multiplex RT - PCR. HA genes of H5N1 viruses from positive samples were amplified by RT -PCR and cloned into pMD18 -T vectors for sequencing respectively. The sequence data were done alignment and phylogenetic analysis with sequences from known reference strains. Results： 18 samples were positive in multiplex RT - PCR and the positive rate was 4.74%. The HA genes from 14 representative positive samples shared 93.1% to 99. 3% identity at nucleotide level and 90. 5% to 99. 1% at amino acid level. 4 different sequences at the cleavage site were obtained and all possessed molecular characteristic of highly pathogenic avian influenza virus. Phylogenetic analysis showed those positive samples could be divided into 5 distinct clades, including clade 1, 2.4, 2. 3.2, 2. 3.4 and 7. Conclusion： HSNI viruses from foreign poultry in boundary region of Yunnan province during 2003 to 2008 had genetic divergence and clade 2. 3.4 viruses were dominant epidemic strains in the region.

关 键 词：禽流感病毒 H5N1亚型 血凝素 进化特征 进化分枝 

分 类 号：S855.3[农业科学—临床兽医学]