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一种用于胃癌 miRNA 分析的简单、灵敏、无标记方法——催化发夹组装辅助分割 G-四联体的程序化设计。

A simple, sensitive and label-free method for miRNA analysis in gastric cancer catalytic hairpin assembly assisted programming of split-G-quadruplexes.

机构信息

Gastroenterology Department, People's Hospital of Chong Qing Liang Jiang New Area, No. 199 Renxing Road, Renhe Street, Yubei District, Chongqing, 401120, China.

出版信息

Anal Methods. 2023 Aug 31;15(34):4236-4242. doi: 10.1039/d3ay00989k.

Abstract

Accurate analysis of miRNA is valuable for the diagnosis of various diseases. Herein, a sensitive and accurate fluorescence method was developed for miRNA detection based on catalytic hairpin assembly (CHA) and split-G-quadruplex (split-G4) based signal reactions. The presence of target miRNA activated the CHA process through unfolding the H1 probe, which could continuously induce the proximity of split-G4. The formed intact G4 can be specifically recognized by the commercial fluorescent dye ThT (thioflavin T), allowing for the highly sensitive, label-free detection of miRNAs. By utilizing split-G4 to generate a signal, the method exhibited a low background signal and a high reliability. In addition, the method is demonstrated to be applied for clinical sample detection, implying its promising prospect for disease diagnosis.

摘要

准确分析 miRNA 对于各种疾病的诊断具有重要意义。本研究基于催化发夹组装(CHA)和分裂 G-四链体(split-G4)信号反应,开发了一种用于 miRNA 检测的灵敏、准确的荧光方法。靶 miRNA 的存在通过展开 H1 探针激活 CHA 过程,从而持续诱导 split-G4 的接近。形成的完整 G4 可以被商用荧光染料 ThT(硫黄素 T)特异性识别,从而实现 miRNA 的高灵敏度、无标记检测。通过利用 split-G4 产生信号,该方法表现出低背景信号和高可靠性。此外,该方法还被证明可用于临床样本检测,表明其在疾病诊断方面具有广阔的应用前景。

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