Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, 221004 Xuzhou, China; Department of Neurology, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002 China.
Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, 221004 Xuzhou, China.
Biosens Bioelectron. 2017 Oct 15;96:62-67. doi: 10.1016/j.bios.2017.04.024. Epub 2017 Apr 19.
We describe a novel label-free and signal-on electrochemical DNA sensing platform via proximity hybridization triggered hemin/G-quadruplex formation based on the direct electron transfer of hemin. The thiolated modified G-DNA1 was first immobilized onto the Au electrode surface. In the presence of target DNA, Y-junction-structure ternary complex can be formed to trigger the proximity assembly of G-DNA1, hemin, and G-DNA2, which leads to the formation of hemin/G-quadruplex for generation an amplified electrochemical signal by differential pulse voltammetry. The observed signal gain was sufficient to achieve a demonstrated detection limit of 54 fM, with a wide linear dynamic range from 10 to 10 M and discriminated mismatched DNA from perfect matched target DNA with a high selectivity. Benefiting from the one step proximity dependent hemin/G-quadruplex formation, the assay method is extremely simple and can be carried out within 40min. The advantages of free of any label conjugation step, and versatility make it a promising candidate for point-of-care testing and commercial application.
我们描述了一种新型的无标记和信号开启的电化学 DNA 传感平台,通过基于血红素直接电子转移的近邻杂交触发血红素/G-四链体形成。首先将巯基化修饰的 G-DNA1 固定在 Au 电极表面。在存在靶 DNA 的情况下,可以形成 Y 型结构的三元复合物,触发 G-DNA1、血红素和 G-DNA2 的近邻组装,导致血红素/G-四链体的形成,通过差分脉冲伏安法产生放大的电化学信号。观察到的信号增益足以实现 54 fM 的检测限,具有从 10 到 10 M 的宽线性动态范围,并具有高选择性区分错配 DNA 和完全匹配的靶 DNA。受益于一步近邻依赖的血红素/G-四链体形成,该测定方法非常简单,可在 40min 内完成。该方法无需任何标记偶联步骤,通用性强,使其成为即时检测和商业应用的有前途的候选方法。