人感染高致病性禽流感病毒A/Fujian/1/2007(H5N1)株基因组序列测定及分析

Genome sequencing and analysis of a high pathogenic avian influenza virus strain A/Fujian/1/2007(H5N1) isolated from a patient in Fujian,China

目的对福建省分离的高致病性禽流感病毒A/Fujian/1/2007(H5N1)株进行全基因组序列测定,了解病毒序列及进化特征。方法SPF鸡胚分离法分离A/Fujian/1/2007(H5N1)株病毒,并提取病毒RNA。病毒RNA经通用引物逆转录后,应用特异性引物分别扩增病毒各节段,产物经TA克隆到载体中进行测序并对结果进行排列、拼接成完整的病毒基因组。分析病毒重要位点特征,并参照已发表的病毒序列,对A/Fujian/1/2007(H5N1)株作系统进化分析。结果应用自行设计的引物,扩增并获得完整的A/Fujian/1/2007(H5N1)株基因组序列。病毒重要位点的分析表明,该株病毒为禽源高致病性禽流感病毒,病毒对达菲敏感而对金刚烷胺类药物则具有耐药性。病毒特征有待进一步的生物学验证。系统进化分析显示,在亚洲各国引起人类感染的高致病性禽流感病毒具有明显的地理分布特征,A/Fujian/1/2007(H5N1)病毒与国内以往毒株属同一进化分支。结论获得人感染高致病性禽流感病毒A/Fujian/1/2007株全基因组序列,了解病毒重要的氨基酸位点特征以及亚洲人感染病毒间的亲缘关系,为进一步探讨福建省禽流感病毒变异、传播机制等奠定基础。

To obtain the genomic sequence of the high pathogenic avian influenza virus （HPAIV） strain A/Fujian/1/ 2007（H5N1） and to explore the feature of the virus and its phylogenie relation with other viruses caused human infection circulating in Asia. Embroynated SPF chicken egg was applied to isolate the A/Fujian/1/2007 （H5N1） virus. The viral RNA was extracted and reverse-transeripted with universal primer, followed by the amplification of viral segments with respective speelfie primers. The amplified products of viral segments were cloned into T-A vector for the sequencing and the results were assembled into a complete genome. The following analysis of important viral amino acid site indicated the A/Fujian/1/2007（H5N1） belongs to avian-original HPAIV and results suggested the virus possesses oseltamivir-sensitivity and amantadine-resistibility that should need further phenotypie test. Phylogenie analysis demonstrates that the HPAIVs caused human infection circulating in different countries or regions in Asia could be clustered into sub-lineages according to their geographical distribution, and the A/Fujian/1/2007（H5N1） could be classified into the sub-lineage consists of the viruses found in human eases previously in mainland, China. The information of viral genome and the important amino acid sites of A/Fujian/1/2007 （H5N1）, and the phylogenie relationship could benefit the further research on the mechanism of HPAIV mutation and transmission in Fujian province.