PCR-荧光探针法在成人HIV/AIDS接受长期高效抗逆转录病毒治疗后的应用研究

HIV-1 provirus in HIV/AIDS patients receiving long-term HAART

目的探讨PCR-荧光探针法检测成人艾滋病病毒感染者和艾滋病患者（HIV/AIDS）接受高效抗逆转录病毒治疗（HAART）后病毒储存库及病毒储存库与治疗效果的关系。方法对91例规范接受HAART治疗5~10年的HIV-1患者采集外周血标本进行横断面研究。通过分支DNA（bDNA）法检测血浆HIV-1RNA病毒载量;用PCR-荧光探针法检测外周血HIV-1DNA水平;用流式细胞术检测外周血CD4＋T细胞数量;对数据进行统计学处理。结果 91例接受5~10年HAART治疗的HIV-1患者外周血HIV-1RNA均低于检测下限（〈50CPs/ml）;细胞中HIV-1前病毒DNA的平均含量为1.77±0.76（lgCPs/106个细胞）;CD4＋T细胞平均数量为（479.56±188.11）个/μl。结论在血浆HIV-1RNA低于检测下限时,PCR-荧光探针法能够检测出经过长期HAART治疗（5~10年）后成人HIV/AIDS的前病毒HIV-1DNA,对于抗病毒治疗疗效的判断以及病情进展的预测都有重要的意义。

Objective To study changes of the virus reservoir in HIV/AIDS patients receiving highly active antiretroviral therapy（HAART）using HIV-1DNA detection kit. Methods A group of 91 chronic HIV-1infected adults who received 5-10 years of HAART treatment,participated in this cross-sectional study.Peripheral whole blood sample was obtained from each patient.The HIV-1RNA viral load was detected by the bDNA method;DNA was extracted using human DNA detection kit.PCR-fluorescent probing was used to detect HIV-1DNA.The T cell counts were obtained by flow cytometry.SPSS software was used for data analysis.Results Among 91 patients with 5-10 years of HAART treatment,plasma HIV-1RNA samples were undetectable;however,HIV-1DNA was detected in the peripheral blood mononuclear cells（PBMCs）of all patients[mean（1.77±0.76）lg copies/million cells]by using HIV-1DNA detection kit;the average number of CD4＋T was（479.56±188.11）/μl. Conclusions HIV-1patients showed significant antiviral and immune reconstitution after long-term treatment with HAART（5-10years）,the HIV-1DNA detection kit can detect the virus reservoir,and HIV-1provirus DNA may play an important role in the efficacy of treatment and monitoring the disease progression.