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目的 :研究HIV 1感染者缺损HIV 1DNA的特性。方法 :用长片段PCR法 (LD PCR)研究分析HIV 1感染者外周血单个核细胞 (peripheralbloodmononuclear,PBMCs)和体外培养感染淋巴细胞中HIV 1基因特征 ,使用位于HIV 1DNA链两端的LTR(U5 )、LTR(R)为引物 ,插入有全长HIV 1基因片段的大肠杆菌质粒PNL4 3等为标准对照 ,并对部分标本克隆测序。结果 :经扩增 9.1kb是LD PCR的主要产物 ,但在 10例HIV 1感染者中有 9例PBMCs检测出大小不一、范围较广的缺失HIV 1基因片段 ,经用基因探针杂交 ,发现近HIV 1基因中心部位缺失频率增加 ,缺失结合点常存在 3～ 4个核苷酸短片段直接重复 ,体外培养中HIV 1基因缺损量减少 ,完整和缺失基因的存在与培养中病毒分离的时间密切相关。结论 :HIV 1感染者PBMCs中存在大量HIV 1基因重组和缺损片段
Objective: To study the characteristics of HIV 1 DNA in HIV-1 infected patients. Methods: The characteristics of HIV-1 gene in peripheral blood mononuclear cells (PBMCs) and in infected lymphocytes of HIV-1 infected individuals were analyzed by long PCR. The LTR (U5) , LTR (R) was used as a primer, and E. coli plasmid PNL4 3 with the full-length HIV 1 gene fragment inserted was used as a standard control. Some of the samples were cloned and sequenced. Results: Amplification of 9.1 kb was the main product of LD PCR. However, in 9 of 10 HIV-1 infected PBMCs, HIV 1 gene fragments with different sizes and wide range were detected. By gene probe hybridization, It was found that the frequency of deletion of HIV 1 gene near the center was increased. The missing 3 to 4 nucleotide fragments were often found in the deletion binding sites. The deletion of HIV 1 gene was reduced in vitro. The presence of complete and deleted genes was separated from the virus in culture Time is closely related. Conclusion: A large number of HIV 1 gene recombinant and defective fragments exist in PBMCs of HIV-1 infected patients