本文選題：HIV-1感染　切入點：基因分型　出處：《南方醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：研究目的了解廣州地區(qū)靜脈吸毒人群和男男同性性行為人群HIV-1基因亞型分布特征和進化關(guān)系,尋找HIV-1的新亞型,探索各亞型可能的流行趨勢;分析P6蛋白氨基酸變異特征及7個氨基酸缺失的流行分布,為艾滋病的治療與預(yù)防提供基礎(chǔ)數(shù)據(jù)。研究方法收集來自廣東省皮膚病防治中心MSM(Men who have sex with men)和IDUs(Intravenous drug users)血液樣本182份,提取病毒核酸,運用巢式RT-PCR法擴增HIV-1三個基因片段p17(670bp),pol(840bp),和gp41(461bp)。將擴增產(chǎn)物純化后測序,構(gòu)建ML系統(tǒng)發(fā)育進化樹,確定樣本基因亞型以及毒株之間的進化關(guān)系。對所獲未知基因亞型URFs(unique recombinant forms)毒株進行近全長基因組序列擴增,分析基因組結(jié)構(gòu)和進化特征。擴增CRF07_BC亞型樣本gag全長序列,將P6蛋白翻譯為氨基酸與HXB2比對,分析蛋白的缺失突變,并結(jié)合數(shù)據(jù)庫中P6序列,分析p6A7(7-aa deletion)在不同人群,時間和地區(qū)的流行分布特征,構(gòu)建系統(tǒng)進化樹。研究結(jié)果樣本人群中,HIV-1基因亞型以CRF01_AE,CRF07_BC和CRF08_BC為主,分別占32.4%,27.5%,17.6%。同時還存在CRF55_01B,B,C和CRF59_01B等亞型以及6%的獨特重組株URF。IDU人群病毒基因亞型分布與MSM人群不同。MSM人群基因亞型以CRF01_AE和CRF07_BC為主,分別占45.5%和30.1%。而IDU人群主要為CRF08_BC(54.3%)和CRF07_BC(22.1%)。進化分析顯示,廣州MSM人群中,CRF07 BC和CRF01_AE序列分別聚集為一或二個毒株簇,新疆,泰國,云南等地IDU和異性性行為人群序列位于外圍。IDU人群中,CRF07_BC序列變異較大,散在分布;CRF01_AE毒株與異性性行為人群聚集成簇,存在密切傳播關(guān)系。無論是MSM人群或IDUs,兩個亞型毒株在各地區(qū)之間呈現(xiàn)交叉分布,沒有地域限制。CRF08_BC流行于IDU人群中,形成特有的廣州簇。在所有亞型中,env和gag的基因距離大于pol。同一基因不同亞型的進化距離不同,subtypeB和CRF01_AE距離較大,CRF07_BC和CRF08_BC居中,CRF55_01B距離最小。研究得到一條來自MSM人群的近全長基因組序列GD698,由CRF07_BC和CRF55 01B重組構(gòu)成,是一種新型的URF。與其他CRF07_BC/CRF55 01B重組株相比,GD698兩個片段均來自MSM人群,有著獨特的重組斷點和鑲嵌結(jié)構(gòu)。另外,在296條HIV-1CRF07_BC亞型毒株中,54.1%的序列含有7個氨基酸的缺失(aa30-36,PIDKELY)。IDU人群p6△7流行率為37.3%,MSM人群為92.3%。IDU人群p6△7序列數(shù)量隨時間而增加。2002年以前約為27.3%,2003-2007年增長為31.4%,到2013年流行率約為66.7%。各地區(qū)流行情況也不一致,以南部較高(68.8%),北部居中(41.2%),西部最低(15.4%)。MSM人群則一直保持較高流行水平,不隨時間或地區(qū)變化。進化分析顯示,MSM人群的p6A7毒株來自其他含p6△7的序列,隨病毒快速傳播,形成單一的毒株簇。而IDU人群p6△7序列起源于沒有缺失的毒株,呈散在分布。研究結(jié)論廣州地區(qū)MSM和IDU人群HIV-1基因亞型分布呈現(xiàn)多樣性,兩人群的基因亞型有所不同,IDU人群主要流行的基因亞型為CRF08_BC,MSM人群主要流行的基因亞型為CRF01_AE。IDU人群中HIV-1序列變異程度大,流行時間長;MSM人群毒株聚集成簇,傳播速度快。MSM人群中首次發(fā)現(xiàn)一株由CRF07 BC和CRF55 01B重組構(gòu)成的新型URF,命名為GD698。CRF07_BCP6蛋白的7個氨基酸缺失毒株的流行情況存在人群、時間、地域差異,廣州流行率較高。
[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.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R512.91;R181.3

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