婚内性传播的HIV准种传播和进化规律研究

发布时间：2018-04-22 13:30

本文选题：艾滋病病毒 + 分子流行病学　；参考：《中国艾滋病性病》2015年05期

【摘要】：目的研究婚内性传播的艾滋病病毒(HIV)准种传播和进化规律,并探讨样本类型(全血、血浆)对分析结果的潜在影响。方法选择传播关系和传播时间明确的4对婚内新近传播HIV夫妻作为研究对象,采集血液样本,分别提取全血、血浆中的总核酸、核糖核酸(RNA),进行HIV基因亚型分析。以HIV-1包膜蛋白(env)基因区为目的片段,分别将总核酸或RNA反转录产物进行巢式聚合酶链反应(PCR)扩增,构建脱氧核糖核酸(DNA)文库,Miseq高通量测序,分析HIV准种分布,计算样本内、夫妻样本间的基因离散率,并做系统进化树分析。结果4对夫妻的HIV基因亚型分别相同。除1对夫妻的血浆样本PCR扩增失败外,其余14份全血、血浆样本成功进行Miseq测序,获得134 135(67 831~178 392)条有效序列,代表101(12~455)个独特准种。多数样本中前20个准种序列的频数占总序列数的比例80%;6/8的受检者全血样本中,第1个优势序列的频数占了90%以上;同一受检者全血样本中高频准种的分布比血浆中更集中。基因离散率、系统进化树分析结果显示,4对夫妻的婚内HIV传播关系都与预期相符,传播方向除1份血浆样本外同样与预期相符。结论 HIV准种的多样性和频数分布有助于了解HIV的传播和进化规律,全血样本所得的HIV感染溯源分析结果比血浆样本更可靠、更实用。
[Abstract]:Objective to study the law of quasi-species transmission and evolution of sexually transmitted HIV (HIV) in marriage, and to explore the potential influence of sample types (whole blood, plasma) on the results of analysis. Methods four newly transmitted HIV couples with definite transmission relationship and time were selected as study subjects. Blood samples were collected, total nucleic acid in whole blood, total nucleic acid in plasma and RNA RNA in ribonucleic acid were extracted, and the subtypes of HIV gene were analyzed. The total nucleic acid or RNA reverse transcription products were amplified by nested polymerase chain reaction (PCR) with the HIV-1 envelope protein env gene region as the target fragments. The HIV-1 DNA library was constructed by high throughput sequencing. The distribution of HIV quasispecies was analyzed, and the distribution of HIV quasispecies was calculated. The genetic dispersion rate between husband and wife samples was analyzed by phylogenetic tree. Results the subtypes of HIV gene were the same in 4 couples. Except for the failure of PCR amplification in the plasma samples of one couple, the other 14 whole blood samples were sequenced by Miseq successfully, and the effective sequence of 134135G 67 831C 178392 was obtained, representing 101C 12455) unique quasispecies. The frequency of the first 20 quasispecies sequences accounted for more than 90% in the whole blood samples of 80 / 6 / 8 of the subjects, and the distribution of high frequency quasispecies in the same sample was more concentrated than that in the plasma. The results of genetic dispersion and phylogenetic tree analysis showed that the marital HIV transmission relationships of the four couples were in accordance with expectations, and the direction of transmission was the same as that of one plasma sample. Conclusion the diversity and frequency distribution of HIV quasispecies are helpful to understand the law of HIV transmission and evolution. The traceability of HIV infection from whole blood samples is more reliable and practical than that of plasma samples.
【作者单位】：中国疾病预防控制中心性病艾滋病预防控制中心;云南省德宏州疾病预防控制中心;山东大学第二医院;
【基金】：国家十二五科技重大专项(2012ZX10001001-001)~~
【分类号】：R512.91

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