长沙地区人腺病毒14、55、57型基因序列进化分析

发布时间：2018-04-09 19:50

本文选题：长沙地区　切入点：人腺病毒　出处：《湖南师范大学》2016年硕士论文

【摘要】：目的:了解2012-2014年长沙地区HAdV的分子流行病学特征;比对分析HAdV的14、55、57型结构蛋白基因的同源性及差异性,为HAdV的防控及进化演变提供实验依据。方法:收集2012年4月至2014年3月因急性下呼吸道感染在湖南师范大学第一附属医院(湖南省人民医院)儿童医学中心住院儿童且经前期实验鉴定为HAdV阳性的鼻咽抽吸物及肺泡灌洗液标本,进行分型;选取其中14、55和57型样本,采用细胞培养技术分离病毒;提取其核酸,通过PCR方法扩增fiber、hexon、penton结构蛋白基因以及14、57型全基因并测序;采用Meg4、Aligmen和DNAman等基因软件对其基因序列进行比对分析并构建进化树。结果:1.本实验成功分型的221例样本,分属于B、C、E三个亚组。其中7型112例(50.68%)、3型22例(9.95%)、14型9例(4.07%)、55型2例(0.90%)、21型1例(0.45%)、1型26例(11.76%)、2型25例(11.31%)、5型7例(3.17%)、6型3例(1.36%)、57型3例(1.36%)、4型11例(4.98%);2.HAdV-57型长沙株的全基因组在结构蛋白基因区有碱基突变,fiber及penton存在个别氨基酸替换和变异,各结构蛋白基因及全基因组与genbank中的样本比对,相似度均为99%;3.hadv-14长沙株的全基因组在结构蛋白基因区无碱基突变,与genbank中近年国内外基因样本同源性高;4.长沙地区hadv-14型和55型的penton基因与郴州、兰州地区的相似度为100%,且与genbank中多个样本的相似度为100%;5.长沙地区hadv-55型的fiber基因与郴州地区的相似度为100%,与genbank中来自中国大陆不同地区及不同时间的多个样本相似度为100%,但与兰州地区的相似度为99%;长沙地区hadv-14型的fiber基因各样本间相似度为100%,与兰州地区的大部分及genbank中多数样本的相似度亦为100%,与郴州及兰州地区的个别样本相似度为99%;6.长沙地区hadv-55型的hexon基因与郴州、兰州地区的相似度为100%,与genbank中多数样本的相似度亦为100%;长沙地区hadv-14型的hexon基因各样本间相似度为100%,与郴州、兰州地区的大部分及genbank中多数国内样本的相似度亦为100%;7.hadv-14型和55型的基因组的主要差异位于hexon基因,差异性达8%,而这两种型别的fiber及penton基因的差异性仅为0-0.5%。结论:本研究期间内,长沙地区hadv主要流行型别较前期有变化,以7型、1型、2型为主要流行株;hadv-14型基因组相对稳定;hadv-57型基因组具有时间及空间进化特性;hadv-55型和14型结构蛋白基因相对稳定。
[Abstract]:Objective: to investigate the molecular epidemiologic characteristics of HAdV in Changsha from 2012 to 2014, and to compare and analyze the homology and difference of the structural protein gene of HAdV type 14555, so as to provide experimental evidence for the prevention and control of HAdV and the evolution of HAdV.Methods: from April 2012 to March 2014, children admitted to the Children's Medical Center of the first affiliated Hospital of Hunan normal University (Hunan people's Hospital) due to acute lower respiratory tract infection and identified as HAdV positive by previous experiments were collected.Aspirates and alveolar lavage fluid,The virus was isolated by cell culture technique, and its nucleic acid was extracted. The gene of fiberine hexonpenton structure protein and the total gene of 1457-type were amplified by PCR and sequenced.Meg4Aligmen and DNAman were used to analyze the gene sequence and construct the evolutionary tree.The result is 1: 1.The 221 samples were divided into three subgroups.And penton have individual amino acid substitution and variation,The similarity of all structural protein genes and the whole genome with the samples in genbank was 993.hadv-14. There was no base mutation in the structural protein gene region of Changsha strain, which had high homology with the domestic and foreign gene samples of genbank in recent years.The similarity between the penton gene of hadv-14 type and 55 type in Changsha and Chenzhou, Lanzhou is 100, and the similarity with multiple samples in genbank is 1005.The similarity between the fiber gene of hadv-55 type in Changsha and Chenzhou is 100. The similarity between fiber gene and genbank samples from different regions and different times in China is 100, but the similarity with Lanzhou is 990.The fiber base of hadv-14 type in Changsha is similar to that of hadv-14 in Changsha.Because the similarity between the samples is 100, the similarity with most of the samples in Lanzhou and genbank is 100, and the similarity between the individual samples in Chenzhou and Lanzhou is 9960.The similarity between the hexon gene of hadv-55 type in Changsha and Chenzhou and Lanzhou is 100, and the similarity with most samples in genbank is 100. The similarity of hexon gene of hadv-14 type in Changsha is 100 and 100 with that of Chenzhou.The similarity of most of the samples in Lanzhou and most domestic samples of genbank is also 1000.The main difference between the genome of 7.hadv-14 and 55 is located in the hexon gene, the difference is up to 8, and the difference between the two types of fiber and penton genes is only 0-0.5.Conclusion: during the period of this study, the main epidemic types of hadv in Changsha area were different from those in the earlier period.The genome of type 7 and type 1 was relatively stable and had the characteristics of temporal and spatial evolution. The structural protein genes of type 14 and type 55 were relatively stable.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R725.1

【相似文献】