Jianhong Zhang , Dan Bai , Guoming Xie , Yaxing Xie , Yu Lin , Yulei Hou , Ying Yu , Yaoyi Zhang , Rong Zhao , Zhongzhong Wang , Luojia Wang , Hui Chen
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引用次数: 0
Abstract
Accurate microRNA (miRNA) detection is pivotal in the diagnosis and monitoring of cancer. Entropy-driven catalysis (EDC) has attracted widespread attention as an enzyme-free, isothermal technique for miRNA detection owing to its inherent simplicity and reliability. However, conventional EDC is a single-output mode, limiting the efficiency of signal amplification. In this study, a novel EDC dual-output mode was employed in conjunction with DNAzyme, resulting in the development of an EDC dual-end DNAzyme (EDC-DED) approach for highly sensitive miRNA detection. In this system, miRNA-21 initiated the EDC reaction, producing a large amount of catalytically active dual-end Mg2+-dependent DNAzyme. The DNAzyme further cleaved the reporter cyclically, generating a notably amplified fluorescence signal. The proposed method achieved a low detection limit of 2 pM. Compared with the traditional EDC single-end DNAzyme (EDC-SED) strategy, the present method exhibited superior amplification efficiency, enhancing detection sensitivity by approximately 46.5-fold. Furthermore, this platform demonstrated ideal specificity, satisfactory reproducibility and acceptable detection capabilities in clinical serum samples. Therefore, the straightforward and convenient strategy is a potential tool for miRNA analysis, which may provide a new perspective for biological analysis and clinical application.
精确的微RNA（miRNA）检测在癌症诊断和监测中至关重要。熵驱动催化（EDC）作为一种无酶、等温的 miRNA 检测技术，因其固有的简便性和可靠性而受到广泛关注。然而，传统的 EDC 是一种单输出模式，限制了信号放大的效率。本研究采用了一种新颖的 EDC 双输出模式与 DNA 酶相结合，从而开发出一种用于高灵敏度 miRNA 检测的 EDC 双端 DNA 酶（EDC-DED）方法。在该系统中，miRNA-21 启动 EDC 反应，产生大量具有催化活性的双端 Mg2+ 依赖性 DNA 酶。DNA 酶进一步循环裂解报告基因，产生显著放大的荧光信号。该方法的检测限低至 2 pM。与传统的 EDC 单端 DNA 酶（EDC-SED）策略相比，本方法的扩增效率更高，检测灵敏度提高了约 46.5 倍。此外，该平台在临床血清样本中表现出理想的特异性、令人满意的重现性和可接受的检测能力。因此，这种简单方便的策略是一种潜在的 miRNA 分析工具，可为生物分析和临床应用提供新的视角。
期刊介绍：
Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome.
Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.
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