摘要

We describe a supersandwich type of electrochemical DNA biosensor based on the use of a glassy carbon electrode (GCE) modified with reduced graphene oxide (rGO) sheets that are decorated with gold nanoparticles (Au NPs). Thiolated capture DNA (probe DNA) was covalently linked to the Au NPs on the surface of the modified GCE via formation of Au-S bonds. In presence of target DNA, its 3' terminus hybridizes with capture probe and the 5' terminus hybridizes with signal probe labeled with Methylene Blue (MB). On increasing the concentration of target DNA, hybridization between signal probe and target DNA results in the formation of three different DNA sequences that form a supersandwich structure. The signal intensity of MB improves distinctly with increasing concentrations of target DNA in the sample solution. The assembling process on the surface of the electrode was studied by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Differential pulse voltammetry (DPV) was used to monitor the hybridization event by measuring the changes in the peak current for MB. Under optimal conditions, the peak currents in DPV for MB linearly increase with the logarithm of target DNA concentration in the range from 0.1 mu M to1.0 fM, with a detection limit of 0.35 fM (at an signal/noise ratio of 3). This biosensor exhibits good selectivity, even over single-base mismatched target DNA.