Sensitive electrochemical biosensor for MicroRNAs based on duplex-specific nuclease-assisted target recycling followed with gold nanoparticles and enzymatic signal amplification.

The detection of sequence-specific microRNAs (miRNAs) is an important factor to the diseases diagnosis. Herein, a triple signal amplification electrochemical biosensor for highly sensitive detection of miRNA-21 was developed based on a duplex-specific nuclease (DSN)-assisted target recycling combined with gold nanoparticles (NPs), horseradish peroxidase (HRP) enzymatic signal amplification. The electrochemical biosensor generated significantly amplified amperometric current changes (Δi) for the detection of miRNA-21 down to 43.3 aM, and Δi was proportional to the logarithm of the concentration of miRNA-21 within the range of 0.1 fM to 100 pM. Meanwhile, the inherent selectivity of the term hairpin capture probe endowed the biosensor with high differentiation of similar miRNAs. The good feasibility of the proposed strategy for cell miRNA detection was confirmed by analyzing miRNA-21 in A549 lysates, which indicates its promising potential in biomedical research and clinical analysis.