基于核酸适配体-荧光染料EvaGreen^(TM)快速检测ATP的研究

Study of rapid detecting ATP based on aptamer and DNA-specific dye EvaGreen^(TM)

目的利用荧光嵌入染料EvaGreenTM在单双链DNA溶液中不同的荧光性质构建基于核酸适配体的ATP检测体系,建立一种简单、高效的检测ATP的新方法。方法利用ATP核酸适配体双链DNA(dsDNA)和核酸绿色荧光染料Eva GreenTM嵌合作用,实现了对ATP的检测。考察了ATP的孵育温度、pH、浓度等因素对荧光强度的影响,并对该方法的特异性进行了验证。结果反应体系在12℃,pH 7.5条件下具有最佳实验效果,在最优条件下,最低能检测10-6mol/L的ATP,并具有较高的特异性。结论本研究为ATP的快速检测提供了一种易于操作、低成本的新方法。

Objective To develop a simple, efficient fluorescence detection technology for ATP. Methods The DNA-interealating dye Eva GreenTM aptamer-ATP complex was used to detect quickly ATP. The effects of temperature, concentration, pH and reaction time were studied. Results The reacting system was that the best temperature and pH were 12 ℃ and pH7.5, The ATP at limitation 10-6 mol/L was detected with optimized conditions. Conclusion This research provides a kind of easy operation and low cost method for ATP rapid detection.