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基于四面体DNA框架的循环信号放大策略用于microRNA的电化学检测

Tetrahedral DNA scaffold-based cyclic signal amplification strategy for electrochemical detection of microRNA
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摘要 基于四面体DNA框架和G-四链体(G4)结构,利用靶标介导的无酶链置换循环信号放大策略,构建了一种电化学传感策略,用于microRNA-21的高灵敏检测。根据靶标(microRNA-21)序列设计四面体DNA框架发夹探针,当靶标存在时,其与四面体DNA框架上的发夹探针互补配对杂交,进而启动3条发夹DNA的链置换循环反应,在电极表面形成大量的“Y”型DNA。每个“Y”型DNA结构中的2个末端均有G4结构,可与血红素(hemin)形成一种具有类辣根过氧化物酶特性的G4/hemin DNAzyme,催化3,3',5,5'-四甲基联苯胺和过氧化氢的反应并产生较强的电流信号,从而用于microRNA-21的检测。当靶标不存在时,链置换反应无法启动,因而电化学信号较弱。结果显示,在1.00 fmol/L~1.00 nmol/L浓度范围内,峰电流差值(各浓度电流值-空白组电流值)与microRNA-21浓度的对数值具有良好的线性关系,线性方程为ΔI=45.04lgc+756.1,线性相关系数R^(2)=0.9987,检测限为17.92 amol/L。该方法有望用于与miRNA-21相关的肿瘤早期诊断的临床应用。 The use of specific biomarkers highly expressed on the surface of extracellular vesicles as targets to identify extracellular vesicles can detect cancer-associated extracellular vesicles. Based on this, an assay for capture and quantification of extracellular vesicles was proposed. By modifying nucleic acid aptamers on monodisperse polystyrene microspheres, specific capture of extracellular vesicles was achieved. The sensitivity of the assay was increased by amplification of the fluorescent signal using an aptamer-triggered hybridization chain reaction. Using this method to detect MCF-7 cell-derived extracellular vesicles under optimized conditions, there was a good linear relationship between the fluorescence change ratio and extracellular vesicle concentration in the range of 1.7×10~3-1.7×10~6 particle/μL, with the correlation coefficient(R^(2)) of 0.9984, and the relative standard deviation(RSD) of 1.6%.
作者 黄静 戴志龙 陈观宇 饶盼详 杨宁 许丽兰 陈敬华 章溪 HUANG Jing;DAI Zhilong;CHEN Guanyu;RAO Panxiang;YANG Ning;XU Lilan;CHEN Jinghua;ZHANG Xi(Department of Clinical Pharmacy and Pharmacy Administration,School of Pharmacy,Fujan Medical University,Fuzhou 350108,China;Department of Pharmaceutical Analysis,School of Pharmacy,Fujian Medical University,Fuzhou 350108,China)
出处 《分析试验室》 EI CAS CSCD 北大核心 2024年第2期274-281,共8页 Chinese Journal of Analysis Laboratory
基金 福建省自然科学基金面上项目(2022J01207)资助。
关键词 微小RNA 四面体DNA框架 循环信号放大 DNAZYME 电化学传感 microRNA tetrahedral DNA scaffold cyclic signal amplification reaction DNAzyme electrochemical biosensing
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