Duplex-specific nuclease assisted miRNA assay based on gold and silver nanoparticles co-decorated on electrode interface

Abstract miRNAs are small non-coding RNAs for gene regulation, which serve as promising biomarkers for the diagnosis of certain diseases. In this contribution, we have proposed a convenient electrochemical biosensing strategy based on the interaction between DNA modified gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs). In principle, citrate capped AuNPs and AgNPs can be co-decorated on the electrode successively. However, with the modification of DNA on AuNPs surface, a strong negative layer is formed. AuNPs@DNA modified electrode could then inhibit subsequent adsorption of AgNPs due to the electrostatic repulsion and steric hindrance effect. As a result, electrochemical response from AgNPs is significantly decreased. On the other hand, in the presence of target miRNA, DNA on AuNPs hybridizes with miRNA and can thus be cyclically digested by duplex-specific nuclease (DSN). Without the shield of DNA, AgNPs can be relaunched at the AuNPs modified electrode. By analyzing the silver stripping peak, highly sensitive detection of miRNA can be achieved. This biosensor exhibits the limit of detection as low as 0.62 fM and a broad linear range from 1 fM to 1 pM. It may hold great potential utility for miRNA assay in the applications of biomedical researches and early clinical diagnosis.