摘要
对2008~2014年收集的中国8省市人呼吸道合胞病毒(Human respiratory syncytial virus,HRSV)A亚型87株代表株进行G蛋白编码基因全长核苷酸序列测定和分析,阐明其核苷酸和氨基酸动态变异特征。根据G蛋白第二高变区的靶基因特征,从2008~2014年中国8省市流行的A亚型人呼吸道合胞病毒中挑选代表株,并采用RT-PCR方法对G蛋白编码基因进行扩增和序列测定,通过Sequencher 5.0、MEGA5.05等生物信息学软件进行序列拼接、比对,分析基因亲缘性和氨基酸变异特点。2008~2014年中国8个代表省市收集的296份HRSVA阳性标本,依据靶基因变异程度及特征共挑选出代表株87株,覆盖2008~2014年流行的各基因型,包括GA5,NA4,NA1,NA3,ON1;对87株代表株进行G蛋白编码基因全长序列测定和同源性分析,核苷酸同源性为88.21%~100%,氨基酸同源性为82.09%~100%,与原型株long株的核苷酸和氨基酸同源性分别为89.22%~91.07%和82.77%~86.82%;变异主要集中在第二高变区,其次在胞浆区和第一高变区。根据G蛋白全长核苷酸序列构建亲缘关系进化树,87株代表株可以分为5个分支,与国际上通用的依据G蛋白第二高变区为靶基因进行的基因分型及进化树分支结果一致。本研究系统分析了2008~2014年中国8省市人呼吸道合胞病毒A亚型G蛋白编码基因的基因特征及氨基酸变异情况,为中国人呼吸道合胞病毒的病原学研究,预防和控制提供了重要的病毒学基础研究数据。
For studying the genetic characteristics and amino acid variations in attachment glycoprotein （G） gene of human respiratory syncytial virus subgroup A （HRSVA） in China, Eighty-seven representative i- solates identified as being positive for HRSVA collected in eight provinces/cities in China from 2008 to 2014 were selected and sequenced according to the collection regions/years and genotypes. The coding re- gion of the HRSV G gene was amplified using reverse transcription-polymerase chain reaction （RT-PCR） and sequenced. Sequences were edited using Sequencher v5. 0 software. Sequence alignment and phyloge- netic trees were built by MEGA v5. 05 software. Eighty-seven HRSVA representative isolates were clus- tered into five genotypes： GAS, NA4, NA1, NA3 and ON1. The nucleotides and amino-acids homology within the 87 sequences was 88.21 %-100 % and 82.09 % -100 %, respectively. The amino-acid homology between the 87 sequences and prototype long strain was 82.77%-86.82%. Amino-acid analysis showed that most of the changes occurred in the second hypervariable region, but variations were also found in the first hypervariable region and cytoplasmic region. Phylogenetic analyses based on the sequences in the en- tire coding region were similar to the tree built according to the second hypervariable region for genoty- ping. In the present study, the genetic characteristics and the amino-acid changes were analyzed systemati- cally using data of HRSVA G gene collected in 7 consecutive years in China's Mainland. These data were used for analyses of the etiology, control and prevention of HRSV infection.
出处
《病毒学报》
CSCD
北大核心
2017年第6期821-828,共8页
Chinese Journal of Virology
基金
国家自然科学基金(项目号:81772195)
题目:我国大陆流行的人呼吸道合胞病毒F蛋白抗原位点动态变异研究
国家重大科技专项课题(项目号:2013ZX10004202)
题目:艾滋病和病毒性肝炎等重大传染病防治~~
关键词
人呼吸道合胞病毒(HRSV)
G蛋白
基因特征
氨基酸
变异
Human respiratory syncytial virus （HRSV）
G protein
Genetic characteristics
Amino acid
Variation
