Anand Sanu K, Ziółkowski Robert
Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Stanisława Noakowskiego 3, 00-664 Warsaw, Poland.
Int J Mol Sci. 2025 Jul 6;26(13):6505. doi: 10.3390/ijms26136505.
DNA methylation, as a critical epigenetic modification, plays a central role in gene regulation and has emerged as a powerful biomarker for early disease diagnostics, particularly in cancer. Owing to the limitations of traditional bisulfite sequencing-such as high cost, complexity, and chemical degradation-electrochemical biosensors have gained substantial attention as promising alternatives. This review summarizes recent advancements in electrochemical platforms for bisulfite-free detection of DNA methylation, encompassing direct oxidation strategies, enzyme-assisted recognition (e.g., restriction endonucleases and methyltransferases), immunoaffinity-based methods, and a variety of signal amplification techniques such as rolling circle amplification and catalytic hairpin assembly. Additional approaches, including strand displacement, magnetic enrichment, and adsorption-based detection, are also discussed. These systems demonstrate exceptional sensitivity, often down to the attomolar or femtomolar level, as well as high selectivity, reproducibility, and suitability for real biological matrices. The integration of nanomaterials and redox-active probes further enhances analytical performance. Importantly, many of these biosensing platforms have been validated using clinical samples, reinforcing their translational relevance. The review concludes by outlining current challenges and future directions, emphasizing the potential of electrochemical biosensors as scalable, cost-effective, and minimally invasive tools for real-time epigenetic monitoring and early-stage disease diagnostics.
DNA甲基化作为一种关键的表观遗传修饰，在基因调控中起着核心作用，并已成为疾病早期诊断的有力生物标志物，尤其是在癌症诊断方面。由于传统亚硫酸氢盐测序存在局限性，如成本高、操作复杂和化学降解等问题，电化学生物传感器作为一种有前景的替代方法受到了广泛关注。本文综述了用于无亚硫酸氢盐检测DNA甲基化的电化学平台的最新进展，包括直接氧化策略、酶辅助识别（如限制性内切酶和甲基转移酶）、基于免疫亲和的方法以及各种信号放大技术，如滚环扩增和催化发夹组装。还讨论了其他方法，包括链置换、磁富集和基于吸附的检测。这些系统具有出色的灵敏度，通常低至阿托摩尔或飞摩尔水平，以及高选择性、重现性和对实际生物样品的适用性。纳米材料和氧化还原活性探针的整合进一步提高了分析性能。重要的是，许多这些生物传感平台已通过临床样本验证，增强了它们的转化相关性。本文最后概述了当前面临的挑战和未来的发展方向，强调了电化学生物传感器作为可扩展、经济高效且微创的工具用于实时表观遗传监测和疾病早期诊断的潜力。
