2009年-2014年云南边境禽流感病毒H5N1亚型神经氨酸酶基因进化分析  

Evolution Analysis of Neuraminidase Genes of Avian Influenza H5N1 Subtype Viruses in the Outside Borders of Yunnan Province During 2009 to 2014

作　　者：曾伟[1,2,3] 刘庆亮[1,2,3] 孔强[1,2,3] 张文东[2] 胡媛媛 胡挺松[3] 赵焕云[2] 段博芳[2] 邱薇[3] 范泉水[3] 张应国[2] 宋建领[5] 张富强[3] 

机构地区：[1]云南农业大学,云南昆明650223 [2]云南省动物疫病预防控制中心,云南昆明650051 [3]成都军区疾病预防控制中心,四川成都610041 [4]云南省农业职业技术学院,云南昆明650031 [5]云南省热带亚热带动物病毒病重点实验室,云南昆明650224

出　　处：《动物医学进展》2015年第7期13-18,共6页Progress In Veterinary Medicine

基　　金：云南省科技计划项目(2012CH002);国家公益性行业专项项目(201103008);军队科技计划项目(CWS12J075;13BJYZ37;A12005);云南省后备人才基金项目(2009CI061)

摘　　要：为了解云南边境禽流感病毒H5N1亚型神经氨酸酶(NA)基因变异及进化特征,2009年-2014年期间在云南边境采集境外家禽和野生鸟类棉拭子样品1 500份,经禽流感H5/N1亚型病毒特异性多重RT-PCR检测,对阳性样品进行病毒NA基因扩增、纯化、克隆和测序,并与已知毒株和参考毒株进行序列比对及系统进化分析。结果从1 500份样品中检出禽流感病毒H5N1亚型阳性样品60份;60阳性样品病毒HA基因测序获得21种NA序列,可划分为4个不同进化分支。NA基因与HA基因呈现不同进化关系;云南边境毒株NA aa275位点未发生变异,因此变异毒株对神经氨酸酶抑制剂Oseltamivir(达菲)类抗病毒药物可能无抗性或耐药性;2009年-2014年期间云南边境H5N1亚型病毒NA间具有遗传差异,NA基因进化分支4已成为当地流行的优势毒株。To elucidate the molecular variation and evolution characteristics of neuraminidase （NA） genes of avian influenza HSN1 subtype viruses in the outside borders of Yunnan province, swab samples were collected from foreign poultry and wild birds in the region and screened by H5/N1 subtype-specific multiplex RT-PCR. The NA genes of H5N1 viruse from the positive samples were amplified, purified, and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis were performed with sequences of the known reference strains. 60 positive samples were found out form 1 500 samples. 21 different of NA sequences were obtained from HA gene from the 60 positive samples and can be divided into 4 distinct clades. The NA genes and Hemagglutinin （HA） genes of HSN1 viruses from the outside borders of Yunnan province showed different relationship of genetic evolution. Yunnan border strain NA aa275 loci did not change, so the mutant strains may have no resistance to neuraminidase inhibitors Oseltamivir （tam- iflu）-antiviral drugs. NA subtype HSN1 viruses from the outside borders of Yunnan province during 2009 to 2014 had genetic divergence, and clade 4 strains had becomed the dominant epidemic strains in the region.

关 键 词：禽流感病毒 H5N1亚型 神经氨酸酶 基因 

分 类 号：S852.659.5[农业科学—基础兽医学]