人感染高致病性H5N6禽流感病毒的分子生物学特征  

Molecular biology characteristics of human infection with a highly pathogenic avian influenza H5N6 virus

作　　者：高敏[1,2,3] 崔大伟[1,2] 杨先知[1,2] 谢国良[1,2] 车飞虎[1,2] 孙海燕[1,2] 陈瑜[1,2] 

机构地区：[1]浙江大学医学院附属第一医院检验科,杭州310003 [2]折江省临床体外诊断技术研究重点实验室,杭州310003 [3]湖州市中心医院检验科,浙江湖州313000

出　　处：《临床检验杂志》2016年第2期156-159,共4页Chinese Journal of Clinical Laboratory Science

基　　金：“十二五”国家科技重大专项(2012ZX10004-210);浙江省医药卫生科技项目(2015KYB149)

摘　　要：目的分析2014-2015年3例新型人感染高致病性禽流感（HPAI）H5N6毒株的血凝素（HA）与神经氨酸酶（NA）基因序列,探讨该病毒的遗传变异及分子生物学特征。方法收集Gen Bank、全球共享禽流感数据倡议组织（GISAID）及浙江大学传染病诊治国家重点实验室提供的H5N6病毒基因序列,利用生物信息软件MEGA6.0及Net Nglyc服务器对基因序列进行遗传进化和分子变异特征分析。结果 3例感染HPAI-H5N6患者毒株的HA与NA氨基酸序列的同源性分别为96.5%～98.7%和90.1%～98.2%;HA氨基酸序列比对分析发现A/Guangzhou/39715/2014（H5N6）和A/Sichuan/26221/2014（H5N6）与A/duck/Eastern China/1111/2011（H5N2）同源性最近,分别为97.7%和98.5%;A/Yunnan/0127/2015（H5N6）与A/muscovy duck/Vietnam/LBM631/2014（H5N1）同源性最近（97.2%）;NA氨基酸序列比对分析发现A/Guangzhou/39715/2014（H5N6）和A/Yunnan/0127/2015（H5N6）与A/swine/Guangdong/K6/2010（H6N6）同源性最近,分别为97.2%和97.6%,A/Sichuan/26221/2014（H5N6）与A/duck/Guangdong/S3073/2010（H6N6）同源性最近（97.4%）;进化树分析显示3例患者毒株处于2个不同的分支;HA基因包含有S137A、T160A等突变,糖基化分析显示有6～7个N-糖基化位点。结论新型人感染HPAI-H5N6病毒是一种重组病毒,其HA基因来源于亚洲禽类H5亚型,NA基因来源于亚洲禽类H6N6;HA基因位点突变及N-糖基化改变可能有助于增强对人类的感染。Objective To investigate the gene sequences of hemagglutinin（ HA） and neuramidinase（ NA） in three novel,highly pathogenic avian influenza（ HPAI） H5N6 virus strains which infected human during 2014-2015,and analyze their genetic variation and molecular biology characteristics. Methods The sequences related to influenza H5N6 virus were collected from Gen Bank,the global initiative sharing avian influenza data（ GISAID） and the State Key Laboratory for Diagnosis and Treatment of Infectious Diseases in Zhejiang University. Then,the heredity evolution and molecular variation characteristics of the sequences were analyzed by MEGA6. 0and Net Nglyc server. Results The homologies of HA and NA amino acid sequences of the three HPAI-H5N6 virus strains（ A / Guangzhou /39715 /2014,A / Sichuan /26221 /2014 and A / Yunnan /0127 /2015） were 96. 5%-98. 7% and 90. 1%-98. 2%,respectively.The sequence alignment analysis of HA amino acid showed that the highest homologies were 98. 5% between A / Sichuan /26221 /2014 and A / duck / Eastern China /1111 /2011（ H5N2）,97. 7% between A / Guangzhou /39715 /2014 and A / duck / Eastern China /1111 /2011（ H5N2）,and 97. 2% between A / Yunnan /0127 /2015 and A / muscovy duck / Vietnam / LBM631 /2014（ H5N1）,respectively.The sequence alignment analysis of NA amino acid showed that the highest homologies were 97. 6% between A / Yunnan /0127 /2015 and A / swine / Guangdong / K6 /2010（ H6N6）, 97. 4% between A / Sichuan /26221 /2014 and A / duck / Guangdong / S3073 /2010（ H6N6）,and 97. 2% between A/Guangzhou/39715 /2014 and A/swine/Guangdong/K6 /2010（ H6N6）,respectively. The phylogenetic trees showed that the three strains were located in two different subclades. The mutation of HA gene included S137 A,T160A and etc.. It was predicted that the amino acid sequences of HA contained six to seven N-glycosylation sites. Conclusion The novel HPAIH5N6 virus infected human is a recombinant virus,and its HA and NA genes are from H5 subtype and H6N6 in Asian poultry,re

关 键 词：禽流感H5N6病毒 血凝素 神经氨酸酶 N-糖基化 

分 类 号：R511.7[医药卫生—内科学]