Electrochemical DNA sandwich biosensor based on enzyme amplified microRNA-21 detection and gold nanoparticles

In this work, a novel electrochemical biosensor for miRNA-21 determination, involving a sandwich hybridization assay onto gold nanoparticles modified pencil graphite electrode (PGE) and enzyme signal amplification was reported. The thiol terminated capture probe 1 (SH-P1) was immobilized on the electrode through AuS interaction. In the presence of target miRNA-21, SH-P1 hybridized with the first part of the target, however, the second part hybridizes with a biotinylated probe P2 (B-P2). Then, a streptavidin-conjugated alkaline phosphatase was immobilized by a specific binding of avidin-B-P2. The enzyme catalyzed the electro-inactive α-naphtyl phosphate to an electro-active α-naphtol. The miRNA-21 detection was achieved through the changes of α-naphtol oxidation signals observed at +0.12 V vs Ag/AgCl with Differential Pulse Voltammetry. Under the optimal detection conditions, the biosensor exhibited selective and sensitive detection with a linear range from 200 pM to 388 nM and the detection limit was 100 pM (10 fmol in 100 μL).