检测HIV-1逆转录酶活性的ELISA方法的建立

Establishment of HIV-1 reverse transcriptase ELISA

目的 建立检测HIV 1逆转录酶活性的ELISA方法。方法 应用包被的模板和逆转录酶将生物素标记的dUTP掺入 ,用辣根过氧化物酶反应系统检测酶活性。结果 建立了检测HIV 1逆转录酶活性的ELISA方法 ,并与同位素掺入检测法进行了比较 ,用ELISA法检测了PFA等药物对HIV 1逆转录酶活性的抑制作用 ,对ELISA法的重复性、稳定性及用于抑制剂研究的特异性和敏感性进行了评价。结论 HIV 1RT活性的ELISA检测法具有简便、快速、重复性好、特异性强的特点 ,适用于抗HIV 1逆转录酶药物的大样品量筛选工作 。

Objective To establish an ELISA method for det ec ting the activity of HIV-1 reverse transcriptase. Methods The template was immobilized onto Covalink microtiter plate, a synthesized bio tinlated dUTP was incorporated by reverse transcriptase. The products were detec ted and quantified using a colorimetric streptavidin-horseradish peroxidase rep orter system. Results We have established a HIV-1 revers e transcriptase ELISA assay and measured the inhibition activity of PFA on HIV- 1 reverse transcriptase, comparing this method with the traditional isotope inco rporation assay. At the same time, we have evaluated the stability, repeatabilit y, specificity and sensitivity of this ELISA method. Conclusion We have established a simple and fast ELISA method for screening and stud y of specific inhibitors of HIV-1 reverse transcriptase. [