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基于适体识别诱导的多 DNA 释放增强的原位猝灭电化学发光生物传感器用于灵敏检测致病菌

An in situ quenching electrochemiluminescence biosensor amplified with aptamer recognition-induced multi-DNA release for sensitive detection of pathogenic bacteria.

机构信息

School of Chemistry and Materials Science, Jiangsu Normal University, Xuzhou, 221116, PR China.

School of Chemistry and Materials Science, Jiangsu Normal University, Xuzhou, 221116, PR China.

出版信息

Biosens Bioelectron. 2022 Jan 15;196:113744. doi: 10.1016/j.bios.2021.113744. Epub 2021 Oct 29.

DOI:10.1016/j.bios.2021.113744
PMID:34736100
Abstract

An in situ quenching electrochemiluminescence (ECL) biosensor sensitized with the aptamer recognition-induced multi-DNA release was designed for pathogenic bacterial detection. Benefitting from the high binding ability of the aptamer to targets and large enrichment capacity of magnetic bead separation, the proposed sensing system not only exhibited outstanding identification to Staphylococcus aureus (S. aureus) among various bacteria, but also released abundant signal transduction DNAs. One S. aureus initiated the dissociation of four kinds of DNA sequences, achieving a one-to-multiple amplification effect. These multi-DNA strands were further hybridized with capture DNA, which were assembled to an electrode modified with Ru(bpy)-conjugated silica nanoparticles (RuSi NPs). Then, glucose oxidase (GOD) was introduced via the functional conjugation of GOD-multi-DNA, leading to the presence of HO by in situ catalysis of GOD on glucose. Relying on the ECL quenching of HO in the Ru(bpy) system, S. aureus was quantified with a linear range from 10 to 10 CFU/mL. In addition, the negative results of non-target bacteria and good recovery efficiency in real samples revealed the system's remarkable selectivity and potential application in infectious food tests.

摘要

基于适配体识别诱导的多 DNA 释放的原位猝灭电化学发光(ECL)生物传感器用于致病性细菌检测。得益于适配体与靶标之间的高结合能力和磁珠分离的大容量富集,所提出的传感系统不仅对各种细菌中的金黄色葡萄球菌(S. aureus)表现出优异的识别能力,而且还释放了丰富的信号转导 DNA。一个 S. aureus 引发了四种 DNA 序列的解离,实现了一到多的放大效应。这些多 DNA 链进一步与捕获 DNA 杂交,组装到修饰有 Ru(bpy)-接枝二氧化硅纳米颗粒(RuSi NPs)的电极上。然后,通过 GOD-multi-DNA 的功能共轭引入葡萄糖氧化酶(GOD),导致 HO 通过 GOD 在葡萄糖上的原位催化而存在。依赖于 Ru(bpy)体系中 HO 的 ECL 猝灭,通过 S. aureus 的 10 至 10 CFU/mL 的线性范围进行定量。此外,非靶标细菌的阴性结果和真实样品中的良好回收率表明该系统具有显著的选择性和在传染性食品检测中的潜在应用。

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