Ji Ze, Cheng Suhang, Li Wensi, Xing Yufei, Tang Zhichao, Zhu Xiang, Wang Dan, Hao Chunyan, Wang Beijuan, Shi Minhua
Department of Respiratory and Critical Care Medicine, Suzhou Kowloon Hospital, Shanghai Jiao Tong University School of Medicine, Suzhou 215028, China.
Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Bioelectrochemistry. 2026 Feb;167:109059. doi: 10.1016/j.bioelechem.2025.109059. Epub 2025 Jul 30.
MicroRNAs (miRNAs) have emerged as critical biomarkers for early cancer diagnosis due to their high specificity and stability in biological fluids. In this study, we report a highly sensitive and specific electrochemiluminescence (ECL) biosensor for the detection of miR-31, a miRNA closely associated with non-small cell lung cancer. The sensing platform integrates a PEI-Ru@Ti₃C₂@AuNPs-modified electrode with a ferrocene-labeled DNA probe (DNA1-Fc) to construct a target-responsive signal-on system. Upon recognition of miR-31, an isothermal strand displacement amplification (ISDA) reaction is initiated, producing abundant double-stranded DNA (dsDNA) that activates the CRISPR/Cas12a complex. The trans-cleavage activity of Cas12a then cleaves the DNA1-Fc probes on the electrode surface, removing the quenching ferrocene moiety and restoring the Ru-based ECL signal. Under optimized conditions, the biosensor exhibited a wide dynamic range from 10 aM to 100 pM and a remarkably low detection limit of 1.67 aM. The system also showed excellent specificity against homologous miRNAs, and its applicability was successfully validated in spiked human serum samples, achieving high recovery and reproducibility. The synergistic combination of nanomaterial-enhanced ECL emission, isothermal nucleic acid amplification, and CRISPR-based enzymatic cleavage provides a powerful strategy for ultrasensitive nucleic acid detection. This work offers a promising approach for early cancer diagnosis and has great potential for clinical translation and point-of-care testing.
微小RNA（miRNAs）由于其在生物体液中的高特异性和稳定性，已成为早期癌症诊断的关键生物标志物。在本研究中，我们报道了一种用于检测miR-31的高灵敏度和特异性的电化学发光（ECL）生物传感器，miR-31是一种与非小细胞肺癌密切相关的微小RNA。该传感平台将聚乙二醇化钌@碳化钛@金纳米粒子修饰电极与二茂铁标记的DNA探针（DNA1-Fc）相结合，构建了一个目标响应信号开启系统。在识别miR-31后，引发等温链置换扩增（ISDA）反应，产生大量双链DNA（dsDNA），激活CRISPR/Cas12a复合物。然后，Cas12a的反式切割活性切割电极表面的DNA1-Fc探针，去除淬灭的二茂铁部分，恢复基于钌的ECL信号。在优化条件下，该生物传感器表现出从10 aM到100 pM的宽动态范围和低至1.67 aM的检测限。该系统对同源微小RNA也表现出优异的特异性，其在加标的人血清样本中的适用性得到成功验证，回收率高且重现性好。纳米材料增强的ECL发射、等温核酸扩增和基于CRISPR的酶切的协同组合为超灵敏核酸检测提供了一种强大的策略。这项工作为早期癌症诊断提供了一种有前景的方法，在临床转化和即时检测方面具有巨大潜力。
