中国HIV-1主要结构基因的序列变异性以及耐药相关性突变分析

发布时间：2018-04-30 05:40

本文选题：HIV-1 + 变异　；参考：《复旦大学》2005年博士论文

【摘要】：第一部分:上海及河南HIV-1分离株p24和gp41基因编码区序列变异性及亚型分析目的了解目前我国上海及河南地区HIV分离株p24蛋白编码区的基因变异性以及亚型的分子流行病学。方法收集上海及河南地区38份已经确诊的HIV-1感染患者的血浆标本并抽提出RNA,其中23份采自于河南,感染途径主要是非法献血;15份采自于上海,主要为经输血制品感染的血友病患者以及经性接触感染者。采用RT-PCR和Nested-PCR的方法扩增gag基因的p24以及env基因的gp41免疫支配区(immunodominant region, IDR),使用DNA测序仪直接测序;应用CLUSTAL X、PHYLIP等软件对序列进行比对及进化树分析。结果亚型分析表明所有样本中81.6%为(31/38)为B亚型,在23例河南患者中全部(100%)为B亚型,;在15例上海患者中8例(53.3%)为B亚型,1例为A亚型,5例为CRF01_AE亚型,1例为CRF02_AG亚型。分别对河南与上海地区的B亚型序列进行分析,与国际共享序列相比,23例河南B亚型的p24编码区中发生核苷酸改变的位点比例为1.6%～4.2%,平均为3.2%;8例上海B亚型核苷酸位点改变的比例为2.0%～3.8%,平均为3.4%。在所有变异位点中均未发现有连续G-A的高度突变。通过对所有的序列进行配对比较计算基因离散率,B亚型内p24区以及gp41 IDR的平均基因离散率分别为3.6%和4.4%。对B亚型gp41IDR进一步分析表明,河南HIV-1分离株之间的平均变异率为3.7%,而上海分离株之间的平均变异率为4.6%。B亚型与其他亚型之间的基因离散率则为14.5%～16.8%。应用CLUSTAL-X分别得出上海及河南地区各自的B亚型共享序列并与国际共享序列进行比对,发现这两个共享序列产生氨基酸变异的比例均为2.2%(5/231),其中有3个变异位点相同,分别为A14P、I91V及E180D,而这两个共享序列之间的差异却极小,仅为0.9%(2/231)。进化树分析亦表明所有样本中大多为B亚型,且所有来自河南的B亚型样本以及大部分上海的B亚型样本多与源自泰国的B’亚型相距较近。结论我国上海及河南地区HIV感染以B亚型较为多见,两地区的B亚型之间有较高的同源性,p24基因仍然较为保守。第二部分:中国部分地区HIV-1耐药相关性突变的研究目的了解我国应用高效抗逆转录病毒治疗(HAART)的HIV感染者中耐药性
[Abstract]:Part one: sequence variation and subtype analysis of p24 and gp41 gene coding region of HIV-1 isolates from Shanghai and Henan Objective to investigate the gene variability and molecular epidemiology of p24 protein coding region of HIV isolates from Shanghai and Henan. Methods 38 plasma samples of patients with HIV-1 infection were collected from Shanghai and Henan, 23 of them were collected from Henan province. 15 samples were collected from Shanghai by illegal blood donation. Hemophilia patients infected with blood transfusion products and infected with sexual contact. RT-PCR and Nested-PCR were used to amplify p24 of gag gene and gp41 dominant region of env gene, and DNA sequencer was used to sequence the sequence directly, and CLUSTAL X-PHYLIP software was used to analyze the sequence and phylogenetic tree. Results the subtype analysis showed that 81.6% of the samples were subtype B, and all of the 23 patients in Henan were B subtype, and 8 out of 15 cases in Shanghai were subtype B and 1 case was subtype A and 5 cases were subtype CRF01_AE. One case was subtype CRF02_AG. The B subtype sequences in Henan and Shanghai were analyzed, respectively. Compared with the internationally shared sequence, the percentage of nucleotide changes in the p24 coding region of Henan B subtype in 23 cases was 1.6 / 4.2, with an average of 3.2 / 8 / 8 / 8 / 8 in Shanghai. The percentage of nucleotide changes in the p24 coding region of Henan B subtype was 2.0 / 3.8 and 3.4 / 3.00, respectively. No continuous G-A mutation was found in all mutation sites. The average gene dispersion rates of p24 region and gp41 IDR in subtype B of gene dispersion were 3.6% and 4.4%, respectively. Further analysis of subtype B gp41IDR showed that the average variation rate among Henan HIV-1 isolates was 3.7, while the average variation rate among Shanghai isolates was 4.6.B subtype and other subtypes, the gene dispersion rate between subtype B and other subtypes was 14.5% and 16.8% respectively. Using CLUSTAL-X, the B subtype shared sequences in Shanghai and Henan were obtained and compared with the international shared sequences. It was found that the amino acid variation rate of the two shared sequences was 2.2 / 231, and 3 of them were the same. The difference between the two shared sequences is very small, which is only 0.920 / 231g / 2. The difference between the two shared sequences is very small, which is A14PU I91V and E180D, respectively, and the difference between the two shared sequences is very small. The phylogenetic tree analysis also showed that most of the samples were subtype B, and most of the samples from Henan province and most of the samples from Shanghai were close to the subtype B 'from Thailand. Conclusion the B subtype of HIV infection is more common in Shanghai and Henan regions of China, and the homology of p24 gene between the B subtypes of the two regions is still relatively conservative. Part two: a study on HIV-1 drug-resistance related mutations in some parts of China Objective to investigate the drug resistance in patients with HIV infected with HAART treated with highly effective antiretroviral therapy (HAART) in China.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R373

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