广州地区静脉吸毒人群和男男同性性行为人群HIV-1分子流行病学特征

发布时间：2018-03-03 22:01

本文选题：HIV-1感染　切入点：基因分型　出处：《南方医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究目的了解广州地区静脉吸毒人群和男男同性性行为人群HIV-1基因亚型分布特征和进化关系,寻找HIV-1的新亚型,探索各亚型可能的流行趋势;分析P6蛋白氨基酸变异特征及7个氨基酸缺失的流行分布,为艾滋病的治疗与预防提供基础数据。研究方法收集来自广东省皮肤病防治中心MSM(Men who have sex with men)和IDUs(Intravenous drug users)血液样本182份,提取病毒核酸,运用巢式RT-PCR法扩增HIV-1三个基因片段p17(670bp),pol(840bp),和gp41(461bp)。将扩增产物纯化后测序,构建ML系统发育进化树,确定样本基因亚型以及毒株之间的进化关系。对所获未知基因亚型URFs(unique recombinant forms)毒株进行近全长基因组序列扩增,分析基因组结构和进化特征。扩增CRF07_BC亚型样本gag全长序列,将P6蛋白翻译为氨基酸与HXB2比对,分析蛋白的缺失突变,并结合数据库中P6序列,分析p6A7(7-aa deletion)在不同人群,时间和地区的流行分布特征,构建系统进化树。研究结果样本人群中,HIV-1基因亚型以CRF01_AE,CRF07_BC和CRF08_BC为主,分别占32.4%,27.5%,17.6%。同时还存在CRF55_01B,B,C和CRF59_01B等亚型以及6%的独特重组株URF。IDU人群病毒基因亚型分布与MSM人群不同。MSM人群基因亚型以CRF01_AE和CRF07_BC为主,分别占45.5%和30.1%。而IDU人群主要为CRF08_BC(54.3%)和CRF07_BC(22.1%)。进化分析显示,广州MSM人群中,CRF07 BC和CRF01_AE序列分别聚集为一或二个毒株簇,新疆,泰国,云南等地IDU和异性性行为人群序列位于外围。IDU人群中,CRF07_BC序列变异较大,散在分布;CRF01_AE毒株与异性性行为人群聚集成簇,存在密切传播关系。无论是MSM人群或IDUs,两个亚型毒株在各地区之间呈现交叉分布,没有地域限制。CRF08_BC流行于IDU人群中,形成特有的广州簇。在所有亚型中,env和gag的基因距离大于pol。同一基因不同亚型的进化距离不同,subtypeB和CRF01_AE距离较大,CRF07_BC和CRF08_BC居中,CRF55_01B距离最小。研究得到一条来自MSM人群的近全长基因组序列GD698,由CRF07_BC和CRF55 01B重组构成,是一种新型的URF。与其他CRF07_BC/CRF55 01B重组株相比,GD698两个片段均来自MSM人群,有着独特的重组断点和镶嵌结构。另外,在296条HIV-1CRF07_BC亚型毒株中,54.1%的序列含有7个氨基酸的缺失(aa30-36,PIDKELY)。IDU人群p6△7流行率为37.3%,MSM人群为92.3%。IDU人群p6△7序列数量随时间而增加。2002年以前约为27.3%,2003-2007年增长为31.4%,到2013年流行率约为66.7%。各地区流行情况也不一致,以南部较高(68.8%),北部居中(41.2%),西部最低(15.4%)。MSM人群则一直保持较高流行水平,不随时间或地区变化。进化分析显示,MSM人群的p6A7毒株来自其他含p6△7的序列,随病毒快速传播,形成单一的毒株簇。而IDU人群p6△7序列起源于没有缺失的毒株,呈散在分布。研究结论广州地区MSM和IDU人群HIV-1基因亚型分布呈现多样性,两人群的基因亚型有所不同,IDU人群主要流行的基因亚型为CRF08_BC,MSM人群主要流行的基因亚型为CRF01_AE。IDU人群中HIV-1序列变异程度大,流行时间长;MSM人群毒株聚集成簇,传播速度快。MSM人群中首次发现一株由CRF07 BC和CRF55 01B重组构成的新型URF,命名为GD698。CRF07_BCP6蛋白的7个氨基酸缺失毒株的流行情况存在人群、时间、地域差异,广州流行率较高。
[Abstract]:Objective: To study drug addicts and MSM HIV-1 genotype distribution and evolutionary relationship in Guangzhou area for vein, new subtype of HIV-1, explore the epidemic trend of each subtype; flow analysis for P6 protein amino acid variation and 7 amino acid deletion distribution, to provide the basic data for the prevention and treatment of AIDS the research methods were collected from central Guangdong province skin disease prevention and control MSM (Men who have sex with men) and IDUs (Intravenous drug users) 182 blood samples, extraction of virus nucleic acid amplification of HIV-1 three gene fragments of p17 by nested RT-PCR method (670bp), pol (840bp), and gp41 (461bp). PCR products were sequenced after purification. The phylogenetic tree of ML system development, determine the evolutionary relationship between sample gene subtypes and strains. The unknown genotype URFs (unique recombinant forms) strains were near full-length gene Sequence analysis of genomic amplification, structural and evolutionary characteristics. Amplification of full-length sequence of CRF07_BC subtype samples gag, P6 and HXB2 protein translated into amino acid alignment analysis of protein deletion mutation, and P6 sequence database, analysis of p6A7 (7-aa deletion) in different populations, the distribution characteristics of time and region, construction of system evolution the tree. The results of the sample population, HIV-1 genotype CRF01_AE, CRF07_BC and CRF08_BC, respectively 32.4%, 27.5%, 17.6%. also has CRF55_01B, B, C and CRF59_01B subtypes and 6% URF.IDU genes among virus strains are unique heavy group distribution and MSM gene among different.MSM subtypes in the crowd CRF01_AE and CRF07_BC, respectively accounted for 45.5% and 30.1%. and IDU population is mainly CRF08_BC (54.3%) and CRF07_BC (22.1%). Phylogenetic analysis showed that the Guangzhou MSM group, CRF07 BC and CRF01_AE sequences were clustered into one or Two strains of Xinjiang, Thailand, Yunnan clusters, such as IDU and heterosexuals sequence located in the periphery of the.IDU population, CRF07_BC sequence variation, scattered distribution; CRF01_AE strains with sex population clusters, there is a close relationship between the spread of either MSM or IDUs. Population, two strains showed cross distribution in different regions, no geographical restrictions.CRF08_BC popular in the IDU population, Guangzhou formed a unique cluster. In all subtypes, different genetic distance Env and gag is greater than pol. the same gene in different subtypes of the evolutionary distance, subtypeB and CRF01_AE distance CRF07_BC and CRF08_BC center, CRF55_01B. We got a minimum distance from the MSM population near full-length genome sequences of GD698, CRF07_BC and CRF55 formed by recombinant 01B, is a new type of URF. CRF07_BC/CRF55 01B and other recombinant strain by GD698 two fragments are from MSM The crowd, with recombination breakpoints and mosaic structure unique. In addition, in the 296 HIV-1CRF07_BC subtypes, 54.1% deletion sequence containing 7 amino acids (aa30-36, PIDKELY).IDU population P6 delta 7 prevalence rate was 37.3%, MSM number of 92.3%.IDU people before the crowd P6 delta 7 time increases in.2002 sequence about 27.3%, 2003-2007 years to 2013 31.4% growth rate is about popular regional 66.7%. epidemic situation is also inconsistent, South high (68.8%), middle (41.2%), North West is the lowest (15.4%).MSM group has maintained a high level of popularity, with time or area changes. The phylogenetic analysis showed that MSM, people from other p6A7 strains containing P6 delta 7 sequences, with the rapid spread of the virus, the formation of a single cluster strains. IDU population P6 delta 7 originated in the sequence no missing strains were scattered. The conclusion of the study of HIV-1 in Guangzhou region MSM and IDU population genotype The distribution of the diversity of two population genetic subtypes in different IDU subtypes in the crowd mainly for CRF08_BC, MSM main group of subtypes of HIV-1 sequence variation degree of the CRF01_AE.IDU population, the popular long time; the MSM population strains clustered together, for the first time found that the new URF was composed of CRF07 BC CRF55 and 01B recombinant spread speed.MSM population, named prevalence was 7 amino acid deletion strains GD698.CRF07_BCP6 protein in the presence of people, time, regional difference, Guangzhou prevalence rate is high.

【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.91;R181.3

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