基于核酸适配子的高灵敏的ATP检测研究

Strategies for high sensitivity detection of ATP based on aptamer

目的:根据ATP核酸适配子与ATP结合的能力强于其互补核酸的原理,构建基于核酸适配子的高灵敏的ATP检测方法。方法:以ATP核酸适配子作为识别分子,对检测的捕获探针固定浓度、杂交时间、ATP反应时间、银染增强时间等进行优化,在优化的反应条件下,对其敏感性、特异性进行评价。结果:在优化的反应条件下,通过对不同浓度的ATP样本进行检测,发现ATP浓度在10-14M至10-9M范围内时,ATP浓度的对数与灰度值的相对强度呈线性关系,相关系数为0.996。并且,在优化的反应条件下,该方法对ATP的检测具有较高的特异性。结论:所构建的基于核酸适配子的ATP检测方法具有灵敏度高、特异性高的优点,可为其他生物分子的检测提供新的思路。

Objective ：According to the principle that the combining capacity of ATP aptamer with ATP is stronger than its complementary nucleic acid ,we built high sensitive detection method for ATP based on aptamer .Methods ：ATP aptamer was taken as recognition molecule ,we optimized reaction conditions of the capture probe fixed concentration , hybridization time ,ATP response time and silver enhancement time , and evaluated the sensitivity and specificity of this method .Results ：Tested the different concentrations of ATP under the optimal conditions ,we found that the relative intensity of the greyscale was proportional to the logarithmic value of ATP concentration over a 5-decade range from 10-14M to 10 -9M with a linear correlation coefficient of 0 .996 .Under the optimal conditions ,the proposed method has high specificity for ATP detection .Conclusion ：This study built a aptamer based ATP detection method with high specificity and high sensitivity ,and can provide a fresh perspective for the detection of other materials .