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目的建立一种合理、详尽、可行的HIV-1基因分型方法。方法采集南昌地区部分男性性接触人群(MSM)人类免疫缺陷病毒(HIV)感染者血浆,测量HIV病毒载量。提取其病毒RNA,设计引物并对HIV的Env区和Gag区进行反转录-聚合酶链反应。对扩增后的c DNA进行测序,比对序列并分别构建Env区和Gag区的分子进化树,确定样本的基因亚型,并分析p V3环氨基酸序列特征。结果本研究的3例样本中,1例样本的HIV病毒载量为43.9 copies/ml,其余2例均<20 copies/ml。3例样本经核酸序列分析,均鉴定为CRF01_AE亚型。结论此HIV-1基因分型方法合理、详细,可为国内HIV分子流行病学研究提供一些参考。
Objective To establish a reasonable, detailed and feasible HIV-1 genotyping method. Methods The plasma of HIV-infected part of men who had sex with men (MSM) in Nanchang area was collected to measure HIV viral load. The viral RNA was extracted, primers were designed and reverse transcription-polymerase chain reaction was performed on HIV Env and Gag regions. The amplified cDNAs were sequenced, the sequences were aligned, and the molecular phylogenetic tree of Env region and Gag region was constructed respectively to determine the gene subtype of the sample, and the amino acid sequence of p V3 loop was analyzed. Results Of the three samples in this study, one had HIV viral load of 43.9 copies / ml and the remaining two had <20 copies / ml. 3 cases of samples by nucleic acid sequence analysis, were identified as CRF01_AE subtype. Conclusion The HIV-1 genotyping method is reasonable and detailed, which can provide some references for HIV molecular epidemiology in China.