| 肖雪,张文东,段博芳,赵焕云,刘庆亮,胡挺松,邱薇,冯子良,郑颖,范泉水,张应国,张富强.云南边境地区禽流感H5N1亚型病毒NS基因进化分析[J].Chinese journal of Epidemiology,2013,34(5):491-495 |
| 云南边境地区禽流感H5N1亚型病毒NS基因进化分析 |
| Genetic evolution of non．structural gene among avian influenza H5；N1 viruses isolated from the boundary of Yunnan province |
| Received:December 24, 2012 |
| DOI：10.3760/cma.j.issn.0254-6450.2014.02.018 |
| KeyWord: 禽流感病毒 H5N1亚型 非结构蛋白l 非结构蛋白2或核输出蛋白 遗传分析 |
| English Key Word: Avian influenza virus H5N1 subtype Non-structural protein 1 Non-structural protein 2 or Nuclear export protein Genetic analysis |
| FundProject:云南省科技计划(2012CH002);国家公益性行业科研专项(201103008) |
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| Abstract: |
| 目的 阐明云南边境地区禽流感H5N1亚型病毒NSl、NS2基因变异特征及遗传进化关系。方法 在云南边境地区采集境外家禽和野生鸟类棉拭子样品，经H5N1亚型特异性多重RT-PCR检测，阳性样品对病毒NS基因进行扩增，克隆Z至pMDl8一T载体测序，获得NSl、NS2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H5N]亚型阳性样品71份，阳性率为5．72％；30份代表性阳性样品病毒NS基因测序获得17种序列，存在3个不同进化(亚)分支(I一1、I一2、Ⅱ)；NSl／NS2基因与病毒血凝素(HA)基因呈现不同进化关系；NSl蛋白涉及核定位信号区、RNA结合区、效应区及其他致病性相关的关键性氨基酸位点存在替代或突变。结论 云南边境地区H5N1亚型病毒NSl／NS2基因具有遗传差异，2010年以来NS基因进化分支I一2、Ⅱ已成为当地流行的优势毒株。 |
| English Abstract: |
| r：ZHANG Fu—qiang，Email：zfq
l 968@yahoo．com．cn
Objective To elucidate the characteristics of variation and the genetic evolution of non．structural protein(NSl．NS2)genes related to avian influenza subtype H5Nl viruses isolated from the boundary region of Yunnanprovince．Methods Swab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5Nl subtype— specific multiplex RT．PCR．The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD 1 8-T vectors for sequencing．The alignment and phylogenetic analysis on those available NS 1．NS2 genes were performed with sequences of the known reference strains．Results 7 1 positive samples were identified from l 240 samples，with the positive rate as 5．72％．Fourteen different NS segment sequences were obrained from 30 representative positive samples and could be divided into 3 distinct clades or sub—clades(I—l，I一2 and II)，by phylogenetic analysis．The NSI／NS2 genes and Hemagglutinin(HA)genes ofH5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics Oil genetic evolution．The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains，effect domain，and other pathogenici哆markers．Conclusion NS genes of H5N1 subtype viruses in boundary region ofYunnan province showed genetic divergence and the virus of elade I一2 and II had become dominant epidemic strains i13．this region since 2010． |
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