| 题名 | 我国云南省HIV 临床株的分离培养及其生物学特性的研究 |
| 作者 | 姚志红 |
| 学位类别 | 博士 |
| 答辩日期 | 2014-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 郑永唐 |
| 关键词 | 云南 HIV 临床株 生物学特性 亚型 生物学嗜性 耐药性 |
| 其他题名 | Isolation and characterization of the HIV-1 isolates in Yunnan province, China |
| 学位专业 | 细胞生物学 |
| 中文摘要 | 我国云南省HIV-1临床株的分离培养及其生物学特性的研究 博士研究生：姚志红（细胞生物学） 导师：郑永唐 研究员 获得性免疫缺陷综合征(Acquired Immune Deficiency Syndrome，AIDS)是由人类免疫缺陷病毒(Human Immunodeficiency Virus，HIV)感染引起的一种免疫缺陷性疾病。该疾病的显著特征是HIV特异性攻击辅助性T淋巴细胞，造成免疫系统功能进行性破坏，导致各种机会性感染和相关肿瘤的发生而致死。目前，我国HIV感染的人数已超过100万人。因此，有效地预防和防治艾滋病的感染已成为当务之急。多项研究表明，云南省是我国HIV主要流行株的起源地。所以，全面、系统和持续地进行云南省HIV分子流行病学研究，对于云南省乃至全国AIDS的预防策略、临床治疗以及抗HIV药物和疫苗的研发具有重要的科学意义。 本研究主要目的是扩充和完善云南省不同地区、不同感染途径的HIV临床病毒株，建立云南省流行的HIV的血浆、核酸、病毒等复合型毒种库。通过对分离培养的HIV临床病毒株进一步开展生物学特性的研究，能够更好地掌握云南省艾滋病的流行特征、传播来源及变异趋势。为艾滋病的预防控制措施提供科学依据、为医学上抗病毒药物的合理使用以及制定有效的抗病毒方案提供指导。HIV-1毒种库的建立也将为研发具有中国特色的抗HIV药物和疫苗及机制的研究奠定基础。 为了开展该研究，2009年-2011年，我们从云南省昆明市收集HIV-1阳性者的调查问卷共3247份，采集HIV-1阳性全血259人份。我们从德宏、保山、开远、昭通和曲靖等地区采集静脉吸毒者（IDUS）的全血共2098人份，其中德宏盈江480人份、保山474人份、开远329人份、昭通581人份，曲靖234人份。昆明来源的3247人已确定为HIV-1阳性。从年龄来看，样本中年龄最小为1岁，最大为88岁。85.2%人年龄在18-50岁之间，表明性活跃者较容易受到感染。从感染途径来看，包括异性接触94.46%（3067/3247）、静脉吸毒2.87%（93/3247）、同性性接触1.35%（44/3247）和母婴传播0.83%（27/3247）。 本研究，我们用HIV、HBV和HCV检测试剂盒检测了云南省不同地区来源的2080名IDUS的病毒感染情况。结果表明：25.5%的人是HIV-1阳性、77.7%的人是HCV阳性、19.2%的人是HBV阳性。15%的人是HIV/HCV共感染、0.3%的人是HIV/HBV共感染、7.8%的人是HBV/HCV共感染、7.1%的人是HIV/HBV/HCV共感染。通过数据分析显示：云南省不同地区HIV、HBV和HCV的流行情况有显著差异。与其他地区相比，无论是HIV、HBV和HCV单感染还是它们之间的共感染情况，保山地区的感染率都是最低的地区。开远地区HIV和HCV的阳性率最高，德宏盈江HBV的阳性率最高。 本研究中，我们采用传统的PBMC共培养的方法和微量全血共培养的方法从我国云南省642人份HIV-1阳性样本中成功分离421株HIV-1临床分离株，分离率为65.57%。其中，采用微量全血共培养的方法成功分离293株，分离率为77.3%。采用PBMC共培养的方法成功分离128株，分离率为49.4%。为了进一步研究分离株的生物学特性，我们从HIV-1原代培养上清中提取病毒基因组RNA，反转录PCR扩增病毒的gag、pol、env区基因并进行序列测定。使用软件Clustal W（http://www.ebi.ac.uk/Tools/msa/clustalw2/）、Bioedit 和MEGA5.0等软件对基因序列特征进行分析。使用simplot软件进行基因序列重组分析。使用Geno2pheno[coreceptor]2.5（http://coreceptor.bioinf.mpi-inf.mpg.de/index.php）进行生物学嗜性的分析。使用HIV Drug Resistance Database（http://hivdb.stanford.edu/）进行耐药性分析。通过分析，我们确定了云南省92株HIV-1临床株的生物学特性。 本研究我们成功获得92株背景清楚的不同亚型、不同生物学嗜性的HIV-1临床分离株。其中，有一株13.CN.iso145病毒株是CRF01_AE亚型、X4嗜性并且具有Y181C突变位点的分离株。为研发具有中国特色的抗HIV-1药物和疫苗及机制的研究奠定了基础。 关键词：云南，HIV-1临床株，生物学特性，亚型，生物学嗜性，耐药性 |
| 英文摘要 | Isolation and characterization of HIV-1 isolates in Yunnan province, China Ph.D candidate: Zhi-Hong Yao (Cell Biology) Directed by Prof. Yong-Tang Zheng （Laboratory of Molecular Immunopharmacology, Kunming Institute of Zoology, graduate school of the Chinese Academy of Sciences,CAS） Acquired immunodeficiency syndrome (AIDS) caused by human immunodeficiency virus (HIV) is an important public health concern. The salient feature of this disease is that HIV specific attack helper T lymphocytes leading to immune system progressive damage and all sorts of opportunistic infections and related tumors. At present, there are one millions HIV-1 infected patients in China, and it is important to prevent and treat HIV infection effectively. A number of research results showed that Yunnan province is the origin site of China's major epidemic strains. Thus, comprehensive, systems, and ongoing HIV-1 molecular epidemiological studies in Yunnan province will play an important role in making an effective prevention strategy to preventing the HIV-1 prevalence in Yunnan province and even the national and developing effective Anti-HIV drugs and vaccines. The main purpose of this study was to improve and expand the HIV-1 reservoir from different areas of Yunnan province. And we hope to establish the virus seed bank of HIV-1 in Yunnan province including plasma, nucleic acid and HIV-1 primary isolates. Based on the research of HIV-1 biological characteristics, we hope to better grasp the AIDS epidemic characteristics in Yunnan province, investigate the source of transmission and the variation trend of HIV-1, which will provide a scientific basis for AIDS prevention and control measures, for the rational use of antiviral drugs on medicine. Setting up viral seed bank will play an important role in developing Anti-HIV drugs and vaccines and providing important information for research. In this study, a total of 3247 questionnaires and 259 whole bloods were collected in Kunming. A total of 2098 whole bloods from IUDs were recruited, with 480 were from Ying jiang, 474 from Baoshan, 329 from Kaiyuan, 581 from Zhaotong, 234 from Qujing. 3247 participants recruited from Kunming were HIV-1 positive patients. 85.2% of the participants aged from 18 to 50, suggested that sexual activity are more susceptible to infection. And among the participants, 94.46% (3067/3247) of them infected HIV-1 through heterosexual contact, 2.87% (93/3247) are intravenous drug use (IDUs), 1.35% (44/3247) through MSM and 0.83% (27/3247) through mother-to-child transmission (MTCT). In our study, we tested the infection status of HIV, HBV and HCV of 2080 intravenous drug users from different regions of Yunnan province using virus detection kit. The results showed that the infection rates among all participants were 25.5% for HIV, 77.7% for HCV, 19.2% for HBV, 15% for HIV/HCV, 0.3% for HIV/HBV, 7.8% for HCV/HBV and 7.1% for HIV/HBV/HCV. HCV is more prevalent than HIV and HBV among all IDUs. The prevalence of virus infection varied widely by regions in Yunnan province of China. It was notable that Baoshan not only had the lowest HIV infection, but also HBV and HCV infection (70.3% and 7.4% respectively). However, Kaiyuan had the highest ratio of HCV (89.1%), while Yingjiang was the most prevalent region of HBV (44/2%) among 5 sites. This study was carried out to isolate and study the biological characteristics of HIV-1 isolates in Yunnan province, China. A total of 421 HIV-1 clinical isolates was successfully cultured from 642 antiretroviral (ART)-na?ve HIV-1-positive individuals recruited from different regions of Yunnan province performed by co-culture of PBMCs/whole blood of HIV-positive patients with PBMCs of HIV-negative individuals. The separation rate was 65.57%. Among them, 293 strains were cultured by the method of Micro-Whole blood, the separation rate was 77.3%, and 128 strains were cultured by the method of PBMC trained, the separation rate was 49.4%. |
| 语种 | 英语 |
| 公开日期 | 2014-06-20 |
| 内容类型 | 学位论文 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7913]  |
| 专题 | 昆明动物研究所_分子免疫药理学 |
| 推荐引用方式 GB/T 7714 | 姚志红. 我国云南省HIV 临床株的分离培养及其生物学特性的研究[D]. 北京. 中国科学院研究生院. 2014. |
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