2009-2011年青岛市甲型H1N1流行性感冒病毒全基因组特征分析  

Analysis of characteristics of whole-genome of influenza A H1N1 virus in Qingdao between year 2009 and 2011

作　　者：汪照国[1] 杨婷婷[1] 柴青[1] 于萍[1] 刘晓琳[1] 段海平[1] 弋英[1] 

机构地区：[1]山东省青岛市疾病预防控制中心微生物与寄生虫检验科,266033

出　　处：《中华预防医学杂志》2013年第12期1105-1109,共5页Chinese Journal of Preventive Medicine

基　　金：青岛市公共领域科技支撑计划（10-3-3-8-1-nsh）

摘　　要：目的 了解2009-2011年青岛地区甲型H1N1流行性感冒病毒（简称流感病毒）的全基因组特征.方法 选取2009-2011年青岛地区流行的甲型H1N1流感病毒35株,提取病毒RNA,应用逆转录聚合酶链反应（RT-PCR）扩增各基因片段,并进行序列测定;用Sequencher软件完成序列拼接.选取GenBank收录的25条全球同期甲型H1N1流感病毒血凝素（HA）基因全长序列作为参照,用Mega 5.0软件对各基因片段进行系统发育分析及分子特征分析;下载2010年8月至2011年3月间全球分离的1068条该型HA序列用于氨基酸变异分析.结果 在HA基因进化树上,2009-2010、2010-2011年监测季毒株各分列4个分支,每监测季各有一个优势分支.2009-2010年监测季优势分离株HA基因核苷酸同源性为99.6％～99.9％,编码氨基酸同源性为99.1％ ～ 99.8％,2010-2011年监测季优势分离株HA基因核苷酸同源性为99.1％ ～ 99.6％,编码氨基酸同源性为98.2％ ～99.1％,两季优势分离株间HA基因核苷酸同源性为98.8％～99.8％,编码氨基酸同源性为98.0％～99.6％.病毒HA蛋白与疫苗株比较,各监测季分别有14、12处氨基酸变异,涉及10个抗原位点及5个阳性选择位点.神经氨酸酶（NA）蛋白247位、274位氨基酸均分别为丝氨酸（247S）、组氨酸（274H）,M2蛋白31位氨基酸均为天冬酰胺（31N）.结论 2009-2011年青岛地区流行的甲型H1N1流感病毒在持续不断地发生变异而产生抗原漂移;毒株全部为烷胺类药物耐药株,但对神经氨酸酶抑制剂敏感.Objective To investigate characteristics of the whole-genome of influenza A H1 N1 virus circulated in Qingdao from year 2009 to 2011. Methods RNA of 35 influenza A H1N1 virus isolates circulated in Qingdao between year 2009 and 2011 was extracted and all segments were amplified by RT-PCR. The sequence was then detected and assembled by software Sequencher. 25 HA full-length sequences published on GenBank were selected as reference. While MEGA 5.0 software package was explored for phylogenetic analysis to characterize the molecular feature with reference to the whole-genome sequence and the hemagglutinin （HA）. 1068 HA sequences of influenza A H1N1 virus isolated worldwide from August 2010 to March 2011 were downloaded for amino acid mutation analysis. Results On the HA genes phylogenetic tree, the virus were separately divided into 4 clades in 2009 -2010 and 2010 -2011 surveillance season, each with a preponderant epidemic clade. The homogeneity of nucleotide and amino acids of HA isolates were 99. 6% -99. 9% and 99. 1% -99.8% respectively in 2009 -2010 surveillance season; 99. 1% - 99.6% and 98.2% - 99. 1% respectively in 2010 - 2011 surveillance season. The homogeneity of nucleotide and amino acids of the preponderant isolates were separately 98. 8% - 99. 8% and 98.0% - 99.6%. Compared with the vaccine strain ,there were separately 14 and 12 variant amino acids of virus HA in the two surveillance season ,involving 10 antigen sites and 5 positive selected sites. The sequence analysis of neuraminidase protein showed that the positions 247,274 presented serine and histidine （ S247, H274 ） respectively. The sequence analysis of M2 protein showed that the isolated A H1N1 viruses presented asparagine in amino acid site 31 （N31）. Conclusion All the A HI N1 influenza virus circulated in Qingdao from year 2009 to 2011 presented continual variation and therefore caused antigenic drift. All the isolations were adamantane-resistance,but susceptible to inhibitors of neruaminidase.

关 键 词：流感病毒A型 基因组 抗原变异 

分 类 号：R373.13[医药卫生—病原生物学]