信号探针和内参比探针结合在一个发夹结构DNA中的高效比率电化学方法,Analytical Chemistry

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信号探针和内参比探针结合在一个发夹结构DNA中的高效比率电化学方法
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-12-16 00:00:00 , DOI: 10.1021/acs.analchem.6b04209
Chunyan Deng ₁ , Xiaomei Pi ₁ , Pin Qian ₁ , Xiaoqing Chen ₁ , Wuming Wu ₂ , Juan Xiang ₁

Affiliation

在这项工作中，通过将二茂铁标记的信号探针（Fc-sP）和亚甲基蓝修饰的内部参比探针（MB-rP）组合在一起，开发了基于双信号标记的发夹结构DNA（dhDNA）的比例探针。发夹结构的DNA。在此基础上，提出了一种用于生物标志物检测的高性能比例电化学方法。与传统的比例式电化学探针相比，这种dhDNA比例式探针将sP和rP整合为一个结构，从而确保了完全相同的修饰条件以及sP和rP在一个传感界面上的相互依赖性。结果，dhDNA比例探针具有消除环境变化干扰的强大能力，这由以下事实证明：表面粗糙度和pH值的变化对传感器的重现性和稳定性没有显着影响。此外，在提出的策略中，Fc-sP对MB-rP的初始比率响应（（I _Fc-sP / I _MB-rP）⁰）是可控制的，并且可以保持为1：1恒定，这有利于信噪比和灵敏度的增加。当将Fc-sP序列设计为粘蛋白1（MUC1）的适体时，带有Au纳米颗粒信号放大的dhDNA比率传感器可通过一步温育程序用于MUC1的灵敏检测。与传统的比例式传感器相比，提出的dhDNA传感器具有更高的重现性，准确性，稳定性，灵敏性和简便性，这对于传感器在各个领域的实际应用开发具有重要意义。

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High-Performance Ratiometric Electrochemical Method Based on the Combination of Signal Probe and Inner Reference Probe in One Hairpin-Structured DNA

In this work, the dual signal-tagged hairpin structured DNA (dhDNA)-based ratiometric probe was developed by the combination of ferrocene-labeled signal probe (Fc-sP) and methylene blue-modified inner reference probe (MB-rP) in one hairpin-structured DNA. On the basis of this, a high-performance ratiometric electrochemical method was proposed for biomarker detection. In contrast to the conventional ratiometric electrochemical probe, this dhDNA ratiometric probe integrated sP and rP into one structure, which ensured the completely same modification condition and the interdependence of sP and rP on one sensing interface. As a result, the dhDNA ratiometric probe possesses a stronger ability to eliminate the disturbance of environmental change, which was proven by the fact that the changes of the surface roughness and pH value had no significant effects on the reproducibility and stability of the sensor. Moreover, in the proposed strategy, the initial ratio responses of Fc-sP to MB-rP ((I_Fc-sP/I_MB-rP)⁰) are controllable and can be kept constant at 1:1, which is favorable for the increase in signal-to-noise ratio and sensitivity. When the sequence of Fc-sP is designed as the aptamer of mucin 1 (MUC1), the dhDNA ratiometric sensor with signal amplification of Au nanoparticles becomes feasible for the sensitive detection of MUC1 by one-step incubation procedure. Compared with the conventional ratiometric sensor, the proposed dhDNA sensor has higher reproducibility, accuracy, stability, sensitivity, and simplicity, which are significant for the development of the sensor in various fields for practical applications.

更新日期：2016-12-16

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