1株H6N6亚型禽流感病毒A/duck/Jiangsu/022/2009的全基因测序及遗传进化分析  

Genome sequencing and phylogenetic analysis of one H6N6 subtype avian influenza virus A/duck/Jiangsu/022/2009

作　　者：顾敏[1] 宋庆庆[1] 李彦芳[1] 徐全刚[1] 彭大新[1] 刘秀梵[1] 

机构地区：[1]扬州大学农业部畜禽传染病学重点开放实验室,扬州225009

出　　处：《中国人兽共患病学报》2010年第7期642-647,共6页Chinese Journal of Zoonoses

基　　金：国家科技重大专项(2008ZX10004-013)

摘　　要：目的对2009年分离自华东地区某活禽交易市场的1株国内未见报道的H6N6亚型禽流感病毒进行了全基因序列的测定,以探讨该病毒各基因片段的可能来源。方法通过常规的血清学试验确定病毒HA亚型,经RT-PCR方法扩增出各基因片段,连接到pGEM-Teasy载体上进行序列测定,利用BLAST进行序列的同源性分析,并结合GenBank中已收录的H6N6亚型流感病毒及其它相关序列进行遗传进化分析。结果 BLAST结果显示该H6N6病毒的HA基因与近年台湾分离株A/duck/Kingmen/E322/2004(H6N2)的核苷酸一致性最高(94.0%),推导的氨基酸剪切位点序列为P-Q-I-E-T-R-G,是典型低致病性禽流感病毒的特征序列;NA基因与华东地区分离株A/duck/EasternChina/160/2002(H4N6)的亲缘关系最近(核苷酸一致性91.0%);而PB1基因与荷兰2003年间分离的H7N7亚型流感病毒关系密切;PA基因则与1999-2000年间意大利分离的H7N1亚型流感病毒亲缘关系较近;NS1蛋白在第218位提前引入终止密码子致使C末端缺失13个氨基酸。结论国内首次分离到的H6N6亚型禽流感病毒的基因组构成较为复杂,但8个基因片段均属于欧亚系,与分离自北美地区的H6N6病毒的遗传距离较远,分别位于不同的进化分支。One H6N6 subtype avian influenza virus,A/duck/Jiangsu/022/2009（Dk/JS/022/09）,which has not reported in China yet,was isolated from a live poultry market in eastern China in 2009.Dk/JS/022/09 was subjected to genome sequencing to investigate the genomic composition.All the complete genome sequences of other H6N6 subtype influenza viruses available in GenBank and some reference sequences were downloaded to conduct the phylogenetic analysis with Dk/JS/022/09.BLAST searches demonstrated that the HA gene had 94.0% nucleotide identity with A/duck/Kingmen/E322/2004（H6N2）isolated from Taiwan,and the amino acid motif of cleavage site between HA1 and HA2 was P-Q-I-E-T-R-G,which was the typical characteristics of the LPAIVs.The NA gene showed close relationship with A/duck/Eastern China/160/2002（H4N6）.However,the PB1 gene displayed high similarity with some H7N7 HPAIVs isolated in Netherlands in 2003 whereas PA gene clustered with some H7N1 subtype AIVs isolated in Italy during the year 1999 to 2000.The NS1 protein had 13 amino acid deleted at the C-termini thus lost the PDZ binding domain.Those results indicated that although the genome composition might be complicated,all the 8 gene segments of Dk/JS/022/09 are attributed to the Eurasian lineage and genetically distant from the other H6N6 viruses recorded in GenBank which belonged to the North-American lineage.

关 键 词：禽流感病毒 H6N6 基因组 进化分析 H7 

分 类 号：R373[医药卫生—病原生物学]