本文選題：HIV-1 + 變異��； 參考：《復(fù)旦大學(xué)》2005年博士論文

【摘要】：第一部分:上海及河南HIV-1分離株p24和gp41基因編碼區(qū)序列變異性及亞型分析 目的 了解目前我國上海及河南地區(qū)HIV分離株p24蛋白編碼區(qū)的基因變異性以及亞型的分子流行病學(xué)。方法 收集上海及河南地區(qū)38份已經(jīng)確診的HIV-1感染患者的血漿標(biāo)本并抽提出RNA,其中23份采自于河南,感染途徑主要是非法獻(xiàn)血;15份采自于上海,主要為經(jīng)輸血制品感染的血友病患者以及經(jīng)性接觸感染者。采用RT-PCR和Nested-PCR的方法擴(kuò)增gag基因的p24以及env基因的gp41免疫支配區(qū)(immunodominant region, IDR),使用DNA測(cè)序儀直接測(cè)序;應(yīng)用CLUSTAL X、PHYLIP等軟件對(duì)序列進(jìn)行比對(duì)及進(jìn)化樹分析。結(jié)果 亞型分析表明所有樣本中81.6%為(31/38)為B亞型,在23例河南患者中全部(100%)為B亞型,;在15例上�；颊咧�8例(53.3%)為B亞型,1例為A亞型,5例為CRF01_AE亞型,1例為CRF02_AG亞型。分別對(duì)河南與上海地區(qū)的B亞型序列進(jìn)行分析,與國際共享序列相比,23例河南B亞型的p24編碼區(qū)中發(fā)生核苷酸改變的位點(diǎn)比例為1.6%～4.2%,平均為3.2%;8例上海B亞型核苷酸位點(diǎn)改變的比例為2.0%～3.8%,平均為3.4%。在所有變異位點(diǎn)中均未發(fā)現(xiàn)有連續(xù)G-A的高度突變。通過對(duì)所有的序列進(jìn)行配對(duì)比較計(jì)算基因離散率,B亞型內(nèi)p24區(qū)以及gp41 IDR的平均基因離散率分別為3.6%和4.4%。對(duì)B亞型gp41IDR進(jìn)一步分析表明,河南HIV-1分離株之間的平均變異率為3.7%,而上海分離株之間的平均變異率為4.6%。B亞型與其他亞型之間的基因離散率則為14.5%～16.8%。應(yīng)用CLUSTAL-X分別得出上海及河南地區(qū)各自的B亞型共享序列并與國際共享序列進(jìn)行比對(duì),發(fā)現(xiàn)這兩個(gè)共享序列產(chǎn)生氨基酸變異的比例均為2.2%(5/231),其中有3個(gè)變異位點(diǎn)相同,分別為A14P、I91V及E180D,而這兩個(gè)共享序列之間的差異卻極小,僅為0.9%(2/231)。進(jìn)化樹分析亦表明所有樣本中大多為B亞型,且所有來自河南的B亞型樣本以及大部分上海的B亞型樣本多與源自泰國的B’亞型相距較近。結(jié)論 我國上海及河南地區(qū)HIV感染以B亞型較為多見,兩地區(qū)的B亞型之間有較高的同源性,p24基因仍然較為保守。 第二部分:中國部分地區(qū)HIV-1耐藥相關(guān)性突變的研究 目的 了解我國應(yīng)用高效抗逆轉(zhuǎn)錄病毒治療(HAART)的HIV感染者中耐藥性
[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.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R373

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 譚建新;HIV-1B'亞型毒株全基因組分子克隆[D];河北大學(xué);2007年

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本文編號(hào)：1823340