Glovi Alessandra, Kalligosfyri Panagiota M, Miglione Antonella, Singh Sima, Iula Gabriella, Giordano Antonio, De Laurentiis Michelino, Cinti Stefano
Scuola Superiore Meridionale, University of Naples "Federico II", Naples, Italy.
Department of Pharmacy, University of Naples Federico II, Naples, Italy.
Discov Oncol. 2025 Jun 6;16(1):1022. doi: 10.1007/s12672-025-02846-z.
Liquid biopsy has emerged as a promising non-invasive, cost-effective and real-time approach for cancer diagnosis and monitoring, allowing for the detection of biomarkers in bodily fluids. Among these, microRNAs (miRNAs) are a valuable choice due to their stability and ability to reflect the tumor's heterogeneity. Concerning triple negative breast cancer (TNBC), an aggressive subtype, several studies have demonstrated consistent upregulation of miRNA-21. Its elevated levels, linked to poor prognosis, make it valuable for early detection, risk stratification, and targeted therapies. While traditional miRNA quantification methods are accurate, they often require complex procedures and skilled personnel, limiting their accessibility in low-resource environments. These challenges can be addressed by electrochemical point-of-care (POC) platforms, inspired by the glucose strip, offering a good alternative by reducing matrix effects, integrating cost-effective and eco-friendly substrates. In this work, miRNA-21 was selectively detected using a complementary DNA probe modified with methylene blue, as a redox mediator, immobilized onto an AuNPs-functionalized, paper-based screen-printed electrode. Significant experimental parameters and sensor's selectivity were carefully evaluated, allowing miRNA-21 detection in both standard solution and human serum with limit of detection (LOD) 1.2 nM and satisfactory repeatability of about 8%. The platform's performance improved tenfold with an external paper-based origami pre-concentration device, enabling pM-level miRNA detection and advancing its potential for real clinical practice applications. The platform is envisioned as a starting point for developing accessible, rapid, cost-effective POC testing, with significant implications for personalized medicine and early TNBC detection.
液体活检已成为一种有前景的非侵入性、经济高效且实时的癌症诊断和监测方法，可用于检测体液中的生物标志物。其中，微小RNA（miRNA）因其稳定性和反映肿瘤异质性的能力而成为一种有价值的选择。关于三阴性乳腺癌（TNBC）这种侵袭性亚型，多项研究已证实miRNA-21持续上调。其水平升高与预后不良相关，这使其在早期检测、风险分层和靶向治疗方面具有重要价值。虽然传统的miRNA定量方法准确，但通常需要复杂的程序和技术人员，限制了其在资源匮乏环境中的可及性。受葡萄糖试纸启发的电化学即时检测（POC）平台可以解决这些挑战，通过减少基质效应、整合经济高效且环保的底物提供了一个很好的替代方案。在这项工作中，使用用亚甲蓝修饰的互补DNA探针作为氧化还原介质，将其固定在金纳米颗粒功能化的纸质丝网印刷电极上，选择性地检测miRNA-21。仔细评估了重要的实验参数和传感器的选择性，能够在标准溶液和人血清中检测miRNA-21，检测限（LOD）为1.2 nM，重复性约为8%，令人满意。使用外部纸质折纸预浓缩装置后，该平台的性能提高了十倍，能够实现pM水平的miRNA检测，并提升了其在实际临床应用中的潜力。该平台被设想为开发可及、快速、经济高效的POC检测的起点，对个性化医疗和早期TNBC检测具有重要意义。
