中文摘要：

目的 建立SYBR Green I荧光定量PCR技术测定HIV-1前病毒载量方法,并以其观察相关病毒进入抑制剂对HIV-1前病毒的作用.方法 选择HIV-1的gag和env保守区域设计引物,建立SYBR Green I荧光定量PCR测定HIV-1前病毒载量的体系,并以其观察6 μg/ml～6 ng/ml的重组病毒巨噬细胞炎症蛋白（rvMIP）、逆转录酶抑制剂（AZT）、膜融合抑制剂（T-20）、rvMIP＋AZT、rvMIP＋T-20对HIV-1前病毒载量的影响.结果 所建立的荧光定量PCR体系的检测下限为101 Copies,其标准曲线的相关系数为0.998 9,斜率为-4.925,截距为35.70;除6 ng/ml的 rvMIP、AZT、T-20外,三种药物处理后HIV-1前病毒载量均显著减小（P〈0.05）,并呈质量浓度依赖性;联合用药者前病毒载量显著低于单用rvMIP者（P〈0.05）,HIV-1前病毒载量rvMIP＋T-20 〈rvMIP＋AZT〈rvMIP〈T-20〈AZT.结论 本研究建立的SYBR Green I荧光定量PCR检测方法具有灵敏、快速、准确及成本低廉等优点;与T-20或AZT联用可增强rvMIP对HIV-1前病毒的抑制作用.

英文摘要：

Objective To set up a SYBR Green I fluorescent quantitative PCR method for the detection of HIV-I provirus viral load, and use this method to study the effect of virus entry inhibitors on the HIV-1 provirus. Methods The SYBR Green I fluorescent quantitative PCR method for the detection of HIV-1 provirus viral load were encompassed the conservative region of gag and env in the HIV-1, the effects to the HIV-1 proviras viral load by 6ug/ml - 6 ng/ml recombinant viral maerophage inflammation protein （ rvMIP）, reverse transcriptase inhibitor （AZT） , membrane infusion inhibitor （ T- 20）, rvMIP ＋ AZT or rvMIP ＋ %20 were detected respectively. Results The detection limit of the fluorescent quantitative PCR was 101 Copies, The correlation coefficient of the standard curves was 0.998 9, the slope was -4.925, and the intercept was 35.70;except for the rvMIP,AZT,T-20（6 ng/ml） ,the HIV-1 provirus viral load decreased after treated by the three drugs （P 〈 0.05 ） , and showed concentration-depended manner;the provirus viral load after treated by drug combination were lower than that treated by rvMIP only （ P 〈 0.05 ）, HIV-1 provirus viral load rvMIP ＋ T-20 〈 rvMIP ＋ AZT 〈 rvMIP 〈 T-20 〈 AZT. Conclusions The SYBR Green I fluorescent quantitative PCR detection method is sensitive, quick, accu- rate and cheap;rvMIP combined with T-20 or AZT can boost up the inhibition of HIV-1 provirus.