Dual-Mode Aptasensor Assembled by a WO₃/Fe₂O₃ Heterojunction for Paper-Based Colorimetric Prediction/Photoelectrochemical Multicomponent Analysis

The programed bimodal photoelectrochemical (PEC)-sensing platform based on DNA structural switching induced by targets binding to aptamers was innovatively designed for the simultaneous detection of mucin 1 (MUC1) and microRNA 21 (miRNA-21). To promote excellent current intensity as well as enhance the sensitivity of aptasensors, the evenly distributed WO3/Fe2O3 heterojunction was prepared as a transducer material, notably reducing the background signal response and extending the absorption of light. The multifunctional paper-based biocathode was assembled to provide a visual colorimetric assay. When introducing the integrated signal probe (ISP) composed of signal probe 1 (sP1) and signal probe 2 (sP2) on paper-based working units modified with gold nanoparticles (AuNPs), recognition sites of two targets were formed. In the presence of MUC1 protein, both sP1 and the target on the working unit were released into the corresponding colorimetric unit because of the DNA specific recognition. The horseradish peroxidase–streptavidin (HRP–SA) carried by free sP1 could oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) to turn a blue-colored oxidized TMB (oxTMB) in the presence of hydrogen peroxide (H2O2), which ultimately gained a higher photocurrent signal. Furthermore, miRNA-21 was modified on another working unit by binding with sP2, leading to changes in the current signal and thus enabling real-time detection of analytes with the assistance of a digital multimeter. The PEC aptasensor offered a wide dynamic range of 10 fg·mL–1–100 ng mL–1 for MUC1 and 0.1 pM–10 nM for miRNA-21, with a low detection limit of 3.4 fg·mL–1 and 36 fM, respectively. It laid the foundation for synchronous detection of multiple analytes and initiated a new way for the enhancement in modern next-generation disease diagnosis.