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基于靶触发杂交链式反应的超灵敏双信号 miRNA 检测方法

Target-triggered hybridization chain reaction for ultrasensitive dual-signal miRNA detection.

机构信息

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China.

State Key Laboratory of Bioelectronics, National Demonstration Center for Experimental Biomedical Engineering Education, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China.

出版信息

Biosens Bioelectron. 2022 Nov 1;215:114572. doi: 10.1016/j.bios.2022.114572. Epub 2022 Jul 16.

Abstract

A signal amplification sensing system with target-triggered DNA cascade reaction combined with dual-signal readout technology was designed for ultrasensitive analysis of miRNA. The highly conductive metal organic frameworks (MOFs) derivative, N-doped carbon dodecahedron (N-PCD) was deposited with gold nanoparticles as the electrode substrate, which could assist the electron transfer between the molecular probe and the electrode surface, and could remarkably enhance electrochemical response. Tetrahedral DNA nanostructure (T4-DNA) with high structural stability and mechanical stiffness was designed to improve the loading capacity and binding efficiency of the target, thus increasing the sensitivity of the system. The non-enzymatic amplification method based on the DNA cascade reaction allows the electrochemical responses from dual signal DNA probes labeled with ferrocene (Fc) and methylene blue (MB), respectively in turn to improve the reliability of detection. Under optimal conditions, the sensor has a linear range of 5-1.0 × 10 fM, and the limit of detection is as low as 1.92 fM and 3.74 fM for Fc and MB labeled probe, respectively. This strategy raises the promising application for the rapid detection of miRNA targets with low abundance in complex biological systems.

摘要

一种基于目标触发的 DNA 级联反应与双信号读取技术的信号放大传感系统被设计用于 miRNA 的超灵敏分析。高导电性金属有机骨架(MOFs)衍生物 N 掺杂十二面体(N-PCD)沉积有金纳米粒子作为电极基底,可辅助分子探针与电极表面之间的电子转移,并能显著增强电化学响应。设计了具有高结构稳定性和机械刚性的四面体 DNA 纳米结构(T4-DNA),以提高目标物的载物量和结合效率,从而提高系统的灵敏度。基于 DNA 级联反应的非酶扩增方法允许分别用二茂铁(Fc)和亚甲基蓝(MB)标记的双信号 DNA 探针进行电化学响应,从而提高检测的可靠性。在最佳条件下,传感器的线性范围为 5-1.0×10 fM,Fc 和 MB 标记探针的检测限分别低至 1.92 fM 和 3.74 fM。该策略为快速检测复杂生物体系中低丰度 miRNA 靶标提供了有前景的应用。

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