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基于量子点的电化学发光和阳极溶出伏安法 DNA 杂交。

Quantum dot-based DNA hybridization by electrochemiluminescence and anodic stripping voltammetry.

机构信息

Key Lab of Analytical Chemistry for Life Science (MOE), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, P. R. China.

出版信息

Analyst. 2010 Jul;135(7):1773-8. doi: 10.1039/c0an00108b. Epub 2010 May 17.

Abstract

Simple and convenient assays with quantum dots (QDs) as the labels for DNA detection are developed. The probe DNA modified with thiol was first immobilized on a pretreated Au electrode, and then the complementary DNA (cDNA) oligonucleotides were hybridized with the immobilized probes by immersing the probe-modified Au electrode into the cDNA oligonucleotide solution. Finally, the avidin-modified QDs were bound to the biosensor in the presence of biotin-modified cDNA. The fabrication process for the biosensor was monitored by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Different from the traditional sandwich-structure strategy, the QDs bind to the target DNA directly via the biotin-avidin-system. By observing the ECL signal and determination of the cadmium component in QDs, the DNA hybridization event was detected by ECL and square wave anodic stripping voltammetric technique (SWASV) respectively. For SWASV detection, the signal linearly increased with the increase of the logarithm of the cDNA concentration over the range of 50 nM-5 microM. The minimum detectable concentration is 50 pM. For ECL, it showed wider linearity range over 5 nM-5 microM and lower detectable concentration of 10 pM. This indicated that the ECL assay could be comparable to the conventional electrochemical assay. Furthermore, this biosensor possesses high selectivity over different sequences of target DNA oligonucleotides.

摘要

开发了以量子点(QD)为标记物用于 DNA 检测的简单便捷的分析方法。首先将巯基修饰的探针 DNA 固定在预处理的 Au 电极上,然后将互补 DNA(cDNA)寡核苷酸通过将探针修饰的 Au 电极浸入 cDNA 寡核苷酸溶液中与固定的探针杂交。最后,在生物素修饰的 cDNA 存在下,将亲和素修饰的 QD 结合到生物传感器上。通过电化学阻抗谱(EIS)和循环伏安法(CV)监测生物传感器的制造过程。与传统的三明治结构策略不同,QD 通过生物素-亲和素系统直接与靶 DNA 结合。通过观察 ECL 信号和 QD 中镉成分的测定,通过 ECL 和方波阳极溶出伏安法(SWASV)分别检测 DNA 杂交事件。对于 SWASV 检测,信号对数与 cDNA 浓度的增加呈线性关系,范围为 50 nM-5 microM。最低检测浓度为 50 pM。对于 ECL,它在 5 nM-5 microM 范围内表现出更宽的线性范围和更低的检测浓度 10 pM。这表明 ECL 测定法可与传统电化学测定法相媲美。此外,该生物传感器对不同靶 DNA 寡核苷酸序列具有高选择性。

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