基于核酸适体的试纸条检测三磷酸腺苷

Determination of adenosine triphosphate by aptamer strips

将核酸适体(Aptamer)的特异性与纳米金颗粒的独特光学性质相结合,制备了一种适用于小分子检测的干式试纸条。该试纸条以修饰功能化核酸适体(PloyT13-Aptamer)的纳米金为识别元件,在控制线(C)上修饰ployA13序列,与识别元件结合以判断试纸条的有效性;在测试线(T)上修饰与Aptamer部分互补的DNA序列,与待测物形成竞争关系,通过T线上纳米金显色的深浅来定性或定量分析待测物浓度。结果表明,优化实验条件下,观察T线颜色变化可实现三磷酸腺苷(ATP)的肉眼定性检测,视觉检出限为10μmol/L。使用Image J软件进行定量分析,试纸条检测范围为10~1000μmol/L,检出限为2.4μmol/L。该试纸条生物传感器可以在10 min内得出检测结果且特异性良好,血清中回收率为104.4%~119.0%,为ATP的现场快速检测提供了一种经济有效的方法。

A dry strip for small molecule detection was prepared by combining specific aptamers and gold nanoparticles(AuNPs)with unique optical properties.The ployA13 sequence was modified on the control line(C)and the PloyT13-Aptamer nanogold was used as the recognition element to judge the validity of the strip.The DNA sequence complementary to aptamer was modified on the test line(T)to form a competitive relationship with the object to be tested.The concentrations of the target is capable of being analyzed qualitatively and quantitatively by the color shade of the AuNPs on the T line.The results exhibited that the color change of the T-line can be observed under optimized conditions,and the visual detection limit of ATP was 10μmol/L.The detection range of the strip was 10-1000μmol/L with a detection limit of 2.4μmol/L,when Image J software was used for quantitative analysis.The strip biosensor can produce results within 10 min with excellent specificity,and the serum recovery rates were 104.4%-119.0%,which provides an economical and effective strategy for rapid detection of ATP in relevant fields.