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基于金和银纳米粒子共同修饰电极界面的双特异性核酸酶辅助 miRNA 分析。

Duplex-specific nuclease assisted miRNA assay based on gold and silver nanoparticles co-decorated on electrode interface.

机构信息

Suzhou Blood Center, Suzhou, 215006, PR China.

Department of Clinical Laboratory, The Affiliated Suzhou Science & Technology Town Hospital of Nanjing Medical University, Suzhou, 215153, PR China.

出版信息

Anal Chim Acta. 2020 Apr 22;1107:23-29. doi: 10.1016/j.aca.2020.01.041. Epub 2020 Jan 20.

Abstract

miRNAs are small non-coding RNAs for gene regulation, which serve as promising biomarkers for the diagnosis of certain diseases. In this contribution, we have proposed a convenient electrochemical biosensing strategy based on the interaction between DNA modified gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs). In principle, citrate capped AuNPs and AgNPs can be co-decorated on the electrode successively. However, with the modification of DNA on AuNPs surface, a strong negative layer is formed. AuNPs@DNA modified electrode could then inhibit subsequent adsorption of AgNPs due to the electrostatic repulsion and steric hindrance effect. As a result, electrochemical response from AgNPs is significantly decreased. On the other hand, in the presence of target miRNA, DNA on AuNPs hybridizes with miRNA and can thus be cyclically digested by duplex-specific nuclease (DSN). Without the shield of DNA, AgNPs can be relaunched at the AuNPs modified electrode. By analyzing the silver stripping peak, highly sensitive detection of miRNA can be achieved. This biosensor exhibits the limit of detection as low as 0.62 fM and a broad linear range from 1 fM to 1 pM. It may hold great potential utility for miRNA assay in the applications of biomedical researches and early clinical diagnosis.

摘要

miRNAs 是一种用于基因调控的小型非编码 RNA,它们可作为某些疾病诊断的有前途的生物标志物。在本研究中,我们提出了一种基于 DNA 修饰金纳米粒子(AuNPs)与银纳米粒子(AgNPs)相互作用的简便电化学生物传感策略。原则上,可以将柠檬酸钠封端的 AuNPs 和 AgNPs 先后共修饰在电极上。然而,由于 AuNPs 表面修饰了 DNA,形成了强负电层。由于静电排斥和空间位阻效应,AuNPs@DNA 修饰电极随后会抑制 AgNPs 的后续吸附。因此,AgNPs 的电化学响应显著降低。另一方面,在存在靶 miRNA 的情况下,AuNPs 上的 DNA 与 miRNA 杂交,因此可以被双链特异性核酸酶(DSN)循环消化。没有 DNA 的保护,AgNPs 可以重新在 AuNPs 修饰的电极上释放。通过分析银剥离峰,可以实现对 miRNA 的高灵敏度检测。该生物传感器的检测限低至 0.62 fM,线性范围从 1 fM 到 1 pM。它在生物医学研究和早期临床诊断中的 miRNA 检测方面具有很大的应用潜力。

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