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A cyclodextrin host-guest recognition approach to a label-free electrochemical DNA hybridization biosensor

机译:无标记电化学DNA杂交生物传感器的环糊精主客体识别方法。

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摘要

A novel label-free electrochemical DNA hybridization biosensor using a β-cyclodextrin/poly(N-acetylaniline)/carbon nanotube composite modified screen printed electrode (CD/PNAANI/CNT/SPE) has been developed. The proposed DNA hybridization biosensor relies on the intrinsic oxidation signals of guanine (G) and adenine (A) from single-stranded DNA entered into the cyclodextrin (CD) cavity. Due to the binding of G and A bases to complementary cytosine and thymine bases in dsDNA, the signals obtained for ssDNA were much higher than that of dsDNA. The synergistic effect of the multi-walled carbon nanotubes provides a significantly enhanced voltammetric signal, and the CD encapsulation effect makes anodic peaks of G and A shift to less positive potentials than that at the bare SPE. The peak heights of G and A signals are dependent on both the number of the respective bases in oligonucleotides and the concentration of the target DNA sequences. Hybridization of complementary strands was monitored through the measurements of oxidation signal of purine bases, which enabled the detection of target sequences from 0.01 to 1.02 nmol μl ~(-1) with the detection limit of target DNA as low as 5.0 pmol μl ~(-1) (S/N = 3). Implementation of label-free and homogeneous electrochemical hybridization detection constitutes an important step toward low-cost, simple, highly sensitive and accurate DNA assay. Discrimination between complementary, noncomplementary, and two-base mismatch targets was easily accomplished using the proposed electrode.
机译:开发了一种新型的无标记电化学DNA杂交生物传感器,该传感器使用β-环糊精/聚(N-乙酰基苯胺)/碳纳米管复合修饰的丝网印刷电极(CD / PNAANI / CNT / SPE)。提出的DNA杂交生物传感器依赖于进入环糊精(CD)腔的单链DNA的鸟嘌呤(G)和腺嘌呤(A)的固有氧化信号。由于G和A碱基与dsDNA中互补的胞嘧啶和胸腺嘧啶碱基结合,因此ssDNA所获得的信号远高于dsDNA。多壁碳纳米管的协同效应提供了显着增强的伏安信号,并且CD封装效应使G和A的阳极峰转移到比裸露SPE处更小的正电位。 G和A信号的峰高取决于寡核苷酸中相应碱基的数目和靶DNA序列的浓度。通过测量嘌呤碱基的氧化信号来监测互补链的杂交,从而能够检测到0.01至1.02 nmolμl〜(-1)的靶序列,而靶DNA的检测限低至5.0 pmolμl〜(- 1)(S / N = 3)。实现无标记且均质的电化学杂交检测是朝着低成本,简单,高度灵敏和准确的DNA分析迈出的重要一步。使用拟议的电极可轻松实现互补,非互补和两碱基错配靶的区分。

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