Yi Wenjuan, Chen Yujie, Gao Caiyun, Qin Qiufang, Qin Heping
Department of Gynecology, Liuzhou People's Hospital, Liuzhou City, Guangxi Zhuang Autonomous Region, 545000, China.
Department of Gastroenterology, Liuzhou People's Hospital, No. 8 Wenchang Road, Central District, Liuzhou City, Guangxi Zhuang Autonomous Region, 545000, China.
Anal Methods. 2025 Jun 19;17(24):5027-5033. doi: 10.1039/d5ay00532a.
MicroRNAs (miRNAs) are pivotal regulators in disease progression and have emerged as significant biomarkers for the early detection, therapeutic intervention, and management of ovarian cancer. The integration of highly sensitive and reliable miRNA detection techniques with ultrasound imaging has the potential to significantly improve the diagnostic accuracy of cancers. In this study, we developed a novel colorimetric approach for the sensitive and reliable detection of miRNAs. This method combines self-priming-mediated DNA polymerization with target recycling and a SYBR Green I (SG)-induced colorimetric response. The sensor operates through a three-stage mechanism: (i) target recognition initiates the formation of a self-priming structure; (ii) an exponential isothermal amplification process, driven by DNA polymerase, facilitates signal amplification; and (iii) a photo-catalyzed color change enables label-free signal generation. Using miRNA-21 as a model target, this approach allows for precise miRNA-21 detection without the need for primers, while the SG-based photo-catalyzed color reaction minimizes background signal interference. The sensor demonstrates the ability to distinguish single-base mismatches in homologous sequences and maintains robust performance in complex biological environments. Furthermore, the sensor has been successfully applied to accurately quantify miRNA-21 levels from constructed samples, highlighting its substantial potential for clinical diagnostics and disease monitoring applications.
微小RNA（miRNA）是疾病进展中的关键调节因子，已成为卵巢癌早期检测、治疗干预和管理的重要生物标志物。将高度灵敏且可靠的miRNA检测技术与超声成像相结合，有可能显著提高癌症的诊断准确性。在本研究中，我们开发了一种用于灵敏且可靠地检测miRNA的新型比色法。该方法将自引发介导的DNA聚合与靶标循环以及SYBR Green I（SG）诱导的比色反应相结合。该传感器通过三阶段机制运行：（i）靶标识别引发自引发结构的形成；（ii）由DNA聚合酶驱动的指数等温扩增过程促进信号放大；（iii）光催化颜色变化实现无标记信号产生。以miRNA-21作为模型靶标，该方法无需引物即可精确检测miRNA-21，而基于SG的光催化颜色反应可将背景信号干扰降至最低。该传感器能够区分同源序列中的单碱基错配，并在复杂生物环境中保持稳健性能。此外，该传感器已成功应用于准确量化构建样本中的miRNA-21水平，凸显了其在临床诊断和疾病监测应用中的巨大潜力。
