LAUSR

MicroRNAs (miRNAs) are considered as significant biomarkers in early diagnosis and treatment of diseases. Herein, an electrochemical biosensor that uses ferrocene (Fc)-functionalized covalent organic frameworks (COFs), a DNA tetrahedron nanostructure (DTN) biosensing interface, and a target catalyzed hairpin assembly (CHA) strategy has been fabricated and successfully developed for the sensitive and specific determination of microRNA-21 (miR-21). The COF served as a linked substrate for immobilization of gold nanoparticles (AuNPs), Fc-COOH, and complementary DNA probe L1 to prepare the electrochemical signal probe COF/Au/Fc/L1, which has a large surface area, extraordinary catalytic properties, and superior biocompatibility to amplify the current signal. The DTN containing a hairpin sequence H1 at one vertex was elaborately designed to construct the biosensing interface; thus, the CHA could be implemented on the electrode surface. In the presence of miR-21, the CHA reaction between H1 and the hairpin H2 was triggered to produce a great number of duplex DNA (H1/H2) with sticky ends. Then, the signal probe COF/Au/Fc/L1 was modified on the electrode surface through the hybridization between L1 and the sticky end of H1/H2, thereby obtaining an amplified Fc current signal. Under optimal conditions, the biosensor showed a wide linear response ranging from 1 fM to 10 nM miR-21, with a low detection limit of 0.33 fM (S/N = 3). Meanwhile, the method showed acceptable accuracy and precision for the determination of miR-21 in human serum.