Near-infrared DNA biosensors based on polysulfonate coatings for the sensitive detection of microRNAs

Abstract

MicroRNAs (miRNAs) play crucial roles in the regulation of immune cell differentiation and the immune response during allergic rhinitis (AR). Studies have shown that miRNA-155 is significantly upregulated in AR pathogenesis. Therefore, miRNA-155 can be used as a biomarker for AR diagnosis. Although fluorescent biosensors based on upconversion nanoparticles (UCNPs) have made significant advances in the detection of miRNAs, developing UCNPs with polymer coatings, efficient surface passivation, and DNA functionalization for hybrid sensing in biological media remains challenging. Herein, hairpin DNA1 (H1) is modified into a thin polysulfonic acid layer on UCNPs by sulfonamide bonds, and the fluorescence of the UCNPs is quenched by the fluorescence resonance energy transfer (FRET) process of BHQ3 carried by H1. When the target miRNA-155 is present, the hairpin structure of H1 is opened, allowing BHQ3 to move away from the UCNP surface, and the fluorescence of UCNP is restored. At the same time, hairpin DNA1 (H2) can combine with H1 to replace the miRNA-155 that is bound to H1 with the help of the opening stem ring structure of H1, and the replaced miRNA-155 can continue to react with H1 to amplify the fluorescence signal. Under the optimal experimental conditions, the linear range of miRNA-155 is 0.01–3 nM, with a detection limit of 1.14 pM. Furthermore, the constructed biosensor has been applied to determine miRNA-155 in serum samples, and the spiked recoveries range from 99.8% to 104.8%, which indicates that the developed assay has potential applications in monitoring allergic rhinitis or other miRNA related diseases.