Ratiometric Fluorescent Biosensor Based on Forster Resonance Energy Transfer between Carbon Dots and Acridine Orange for miRNA Analysis

[Image: see text] The expression level of miRNA is highly correlated with the pathological process of malignant tumors. Therefore, the abnormal expression of miRNA in serum is considered as reliable evidence for the existence of tumor cells. Here, a ratiometric fluorescent biosensor based on the Forster resonance energy transfer between fluorophores is proposed for detecting colorectal cancer-specific miRNA (miR-92a-3p). The miRNA in serum was first isolated by carboxyl-modified SiO(2) microspheres. Then, the addition of miRNA to the detection system resulted in the distance change between the donor acridine orange (AO) and the acceptor fluorescent carbon dots (CDs), which made the fluorescence signal change. The physicochemical properties, especially the fluorescence characteristics of CDs and AO, which enabled the ratiometric fluorescence detection, were comprehensively studied. The ratiometric fluorescent biosensor could detect miRNA in the concentration range of 1–9 nM and showed a detection limit of 0.14 nM. Moreover, the ratiometric fluorescent biosensor exhibited high selectivity for the target miRNA. The validity of the ratiometric fluorescent biosensor was also verified using the serum sample, demonstrating its potential for enzyme-free miRNA analysis.