Liu Fanglei, Chen Luqing, Zhu Liuhui, Liu Jihan, Li Min, Chen Yao, Yang Guohai, Qu Lulu
School of Chemistry & Materials Science, Jiangsu Normal University, Xuzhou 221116, P. R. China.
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, P. R. China.
Anal Chem. 2025 Jul 15;97(27):14377-14387. doi: 10.1021/acs.analchem.5c01481. Epub 2025 Jun 30.
Inflammatory bowel disease (IBD) has proven to be a critical global health problem characterized by severe life-threatening complications; thus, the development of noninvasive, reliable, and cost-effective diagnostic methods remains an urgent clinical need. Herein, a novel photoelectrochemical (PEC) and surface-enhanced Raman scattering (SERS) dual-mode platform was successfully developed for ultrasensitive detection of IBD-associated biomarkers, matrix metalloproteinases-9 (MMP-9), and intestinal alkaline phosphatase (IAP). A bifunctional covalent organic frameworks/MXene-Au substrate was synthesized with excellent PEC and SERS properties. An aptamer-based amplification strategy was first employed for MMP-9 detection, which was also the basis for the detection of IAP. The magnetic bead-conjugated double-stranded DNA was then designed to generate activator DNA in the presence of IAP, which activated the -cleavage activity of the CRISPR-Cas12a system. The resultant Cas12a specifically cleaved the electrode-immobilized single-stranded DNA (ssDNA), triggering the release of methylene blue as a dual-signal reporter, thereby enabling synchronized PEC-SERS detection for MMP-9 and IAP. The biosensor exhibited a wide linear range with detection limits of 0.074 pg/mL (PEC) and 0.016 pg/mL (SERS) for MMP-9, and 0.38 pg/mL (PEC) and 0.16 pg/mL (SERS) for IAP, respectively. Significantly, clinical validation was performed using a murine IBD model and human intestinal inflammation specimens, confirming the practical utility of the PEC-SERS platform. This study establishes a robust dual-mode biosensing strategy with multicomponent detection, enabling advanced biological analysis and precision health monitoring.
炎症性肠病（IBD）已被证明是一个严重的全球健康问题，其特征是存在严重的危及生命的并发症；因此，开发无创、可靠且具有成本效益的诊断方法仍然是临床的迫切需求。在此，成功开发了一种新型的光电化学（PEC）和表面增强拉曼散射（SERS）双模式平台，用于超灵敏检测IBD相关生物标志物基质金属蛋白酶-9（MMP-9）和肠碱性磷酸酶（IAP）。合成了具有优异PEC和SERS性能的双功能共价有机框架/MXene-Au基底。首次采用基于适配体的扩增策略检测MMP-9，这也是检测IAP的基础。然后设计了与磁珠偶联的双链DNA，以便在IAP存在时产生激活剂DNA，从而激活CRISPR-Cas12a系统的切割活性。产生的Cas12a特异性切割固定在电极上的单链DNA（ssDNA），触发亚甲基蓝作为双信号报告分子的释放，从而实现对MMP-9和IAP的同步PEC-SERS检测。该生物传感器对MMP-9的线性范围宽，检测限分别为0.074 pg/mL（PEC）和0.016 pg/mL（SERS），对IAP的检测限分别为0.38 pg/mL（PEC）和0.16 pg/mL（SERS）。重要的是，使用小鼠IBD模型和人类肠道炎症标本进行了临床验证，证实了PEC-SERS平台的实际应用价值。本研究建立了一种强大的多组分检测双模式生物传感策略，能够进行先进的生物分析和精准健康监测。
