An electrochemical biosensor for microRNA was constructed on the basis of direct growth of electroactive Prussian Blue (PB) on graphene oxide (GO). A mercapto-modified probe DNA that is complementary to the hepatocellular carcinoma biomarker microRNA-122 was firstly anchored on a gold electrode (AuE). Then, GO (with its large surface and multiple active sites) was adsorbed on probe DNA through π-interaction. Subsequently, the PB nanoparticles were directly grown on GO via alternative dipping the electrode in solutions of FeCl3 and hexacyanoferrate(III). Upon incubation of the resulting electrode with a solution of microRNA-122, the probe DNA on the electrode interacts with microRNA-122 to form a rigid duplex. This results in the release of electroactive PB/GO from the sensing interface and a decrease in current, typically measured at 0.18 V (vs. Ag/AgCl (3 M KCl)). The sensor covers the 10 fM to 10 nM microRNA-122 concentration range and has a 1.5 fM detection limit. The method was successfully applied to the determination of microRNA-122 in real biological samples. Graphical abstract Graphene oxide with in-situ grown Prussian Blue is applied as an electrochemical probe for the analysis of microRNA-122.
Keywords: Atomic force microscopy; Base-pairing principle; Liver injury biomarker; Nanosize effect; Scanning electron microscopy; UV-vis spectra; π-stacking.