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基于杂交链式反应和原子转移自由基聚合的miRNA-21高灵敏检测

Highly sensitive detection of miRNA-21 based on hybridization chain reaction and atom transfer radical polymerization.

作者信息

Zhou Liang, Yu Shuaibing, Tian Zhidan, Kong Jinming, Zhang Xueji

机构信息

School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, 210094, Jiangsu, China.

Department of Pathology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, Jiangsu, China.

出版信息

Anal Chim Acta. 2025 Oct 1;1369:344356. doi: 10.1016/j.aca.2025.344356. Epub 2025 Jun 22.

Abstract

BACKGROUND

Early cancer diagnosis is crucial for improving therapeutic outcomes and survival rates. Research in molecular oncology has shown that tumor-associated biomarkers, particularly miRNAs, are promising key indicators for early detection. However, traditional miRNA detection methods are hindered by complex procedures, insufficient sensitivity, and high costs, limiting their clinical application. Thus, there is an urgent need to develop sensitive, simple, and cost-effective detection platforms.

RESULTS

This study introduces a novel biosensing platform combining Hybridization Chain Reaction (HCR) and Atom Transfer Radical Polymerization (ATRP) for ultrasensitive electrochemical detection of miRNA-21. HCR-driven DNA probe hybridization facilitates target miRNA recognition, while ATRP-mediated signal amplification further enhances the electrochemical response. The HCR mechanism generates multiple initiator anchoring sites via programmable DNA self-assembly, enabling covalent immobilization of ATRP initiators (α-bromophenylacetic acid, BPAA). The ATRP process facilitates the growth of electroactive polymers on the electrode surface, significantly enhancing signal stability and sensitivity. The biosensor exhibits exceptional sensitivity for miRNA-21 detection, with a linear range from 100 pM to 10 aM and a low detection limit of 4.96 aM, outperforming most existing electrochemical methods. Furthermore, the sensor demonstrates strong anti-interference in serum analysis, with a recovery rate of 98.48 %-105.22 %.

SIGNIFICANCE

The HCR-ATRP dual signal amplification strategy offers a robust, cost-effective, and scalable solution for early cancer screening. Its performance in miRNA-21 detection demonstrates exceptional sensitivity, strong anti-interference, excellent stability, and ease of use. This not only highlights its superior detection capability but also underscores the platform's broad potential in identifying other cancer-related biomarkers.

摘要

背景

早期癌症诊断对于改善治疗效果和提高生存率至关重要。分子肿瘤学研究表明,肿瘤相关生物标志物,特别是微小RNA(miRNA),是早期检测的有前景的关键指标。然而,传统的miRNA检测方法存在操作复杂、灵敏度不足和成本高的问题,限制了它们的临床应用。因此,迫切需要开发灵敏、简单且经济高效的检测平台。

结果

本研究引入了一种将杂交链式反应(HCR)和原子转移自由基聚合(ATRP)相结合的新型生物传感平台,用于超灵敏电化学检测miRNA-21。HCR驱动的DNA探针杂交有助于识别目标miRNA,而ATRP介导的信号放大进一步增强了电化学响应。HCR机制通过可编程的DNA自组装产生多个引发剂锚定位点,实现ATRP引发剂(α-溴苯乙酸,BPAA)的共价固定。ATRP过程促进了电极表面电活性聚合物的生长,显著提高了信号稳定性和灵敏度。该生物传感器对miRNA-21检测表现出卓越的灵敏度,线性范围为100 pM至10 aM,检测限低至4.96 aM,优于大多数现有电化学方法。此外,该传感器在血清分析中表现出很强的抗干扰能力,回收率为98.48%-105.22%。

意义

HCR-ATRP双信号放大策略为早期癌症筛查提供了一种强大、经济高效且可扩展的解决方案。其在miRNA-21检测中的性能表现出卓越的灵敏度、强大的抗干扰能力、出色的稳定性和易用性。这不仅突出了其优越的检测能力,也强调了该平台在识别其他癌症相关生物标志物方面的广阔潜力。

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