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基于被核苷酸包裹的银纳米簇探针的痕量和无标记 microRNA 检测。

Trace and label-free microRNA detection using oligonucleotide encapsulated silver nanoclusters as probes.

机构信息

Research Center for Bioengineering and Sensing Technology, University of Science & Technology Beijing, Beijing 100083, People's Republic of China.

出版信息

Anal Chem. 2012 Oct 16;84(20):8670-4. doi: 10.1021/ac301860v. Epub 2012 Sep 28.

DOI:10.1021/ac301860v
PMID:22985191
Abstract

A simple, sensitive, and label-free method for microRNA (miRNA) biosensing was described using oligonucleotide encapsulated silver nanoclusters (Ag-NCs) as effective electrochemical probes. The functional oligonucleotide probe integrates both recognition sequence for hybridization and template sequence for in situ synthesis of Ag-NCs, which appears to possess exceptional metal mimic enzyme properties for catalyzing H(2)O(2) reduction. The miRNA assay employs gold electrodes to immobilize the molecular beacon (MB) probe. After the MB probe subsequently hybridizes with the target and functional probe, the oligonucleotide encapsulated Ag-NCs are brought to the electrode surface and produce a detection signal, in response to H(2)O(2) reduction. An electrochemical miRNA biosensor down to 67 fM with a linear range of 5 orders of magnitude was obtained. Meanwhile, the MB probe allows the biosensor to detect the target with high selectivity. The Ag-NCs-based approach provides a novel avenue to detect miRNA with high sensitivity and selectivity while avoiding laborious label and signal amplification. It is convinced that rational introduction of signal amplification strategy to the Ag-NCs-based bioanalysis can further improve the sensitivity. To our best knowledge, this is the first application of the electrocatalytic activity of Ag-NCs in bioanalysis, which would be attractive for genetic analysis and clinic biomedical application.

摘要

一种简单、灵敏且无需标记的 miRNA(microRNA)生物传感方法,使用被寡核苷酸包裹的银纳米簇(Ag-NCs)作为有效的电化学探针。该功能化的寡核苷酸探针整合了杂交的识别序列和原位合成 Ag-NCs 的模板序列,似乎具有出色的类金属酶特性,可催化 H(2)O(2)还原。miRNA 测定采用金电极固定分子信标(MB)探针。随后,MB 探针与靶标和功能探针杂交后,将被寡核苷酸包裹的 Ag-NCs 带到电极表面,并产生检测信号,以响应 H(2)O(2)还原。该电化学 miRNA 生物传感器的检测下限低至 67 fM,线性范围为 5 个数量级。同时,MB 探针使生物传感器能够高选择性地检测靶标。基于 Ag-NCs 的方法提供了一种高灵敏度和选择性检测 miRNA 的新途径,同时避免了繁琐的标记和信号放大。可以相信,通过合理引入信号放大策略,基于 Ag-NCs 的生物分析可以进一步提高灵敏度。据我们所知,这是首次将 Ag-NCs 的电催化活性应用于生物分析,这将对基因分析和临床生物医学应用具有吸引力。

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