HIV-1前病毒基因扩增诊断早期HIV感染的研究

Study on the early diagnosis of HIV by amplification of HIV pro-virus DNA.

目的建立HIV前病毒核酸的检测方法,并探讨其早期诊断断HIV-1感染的效果。方法从HIV1+2蛋白印迹法确定HIV抗体阳性、并经病毒载量检测阳性的HIV感染者血浆中提取基因组DNA,应用合成HIV-1基因组gag区九条引物随机组合RT-PCR扩增HIVgag区核酸片断,筛选扩增灵敏度最高的三个扩增片断。其引物组合扩增样本核酸PCR产物经凝胶电泳观察判断结果。结果该方法的敏感性为94·17%,特异性为100%,三个片断扩增灵敏度分别为76·67%、87·5%、69·17%。结论PCR扩增HIV前病毒保守区多个基因片断,具有较高的灵敏度和特异度,方法简单,可以应用到HIV感染的早期诊断中。

Objective To establish a method for detection of HIV-1 pro-virus DNA and early infection. Methods Nine primers from HIV-1 nucleic acid sequence gag region were synthesized . The samples confirmed to be positive by western blotting and virus loading were selected . HIV gag region fragments were amplified by RT-PCR and three most sensitive fragments were screened and used for amplification of HIV-1 DNA . The PCR product was determined by gel electrophoresis. Results The sensitivity of this method was 94.17% ad the specificity was 100%. The sensitivity of the three selected fragments were 76.67%, 87.50% and 69.17%. Conclusion This method is highly sensitive and specific an can be used for early diagnosis of HIV-1 infection .