Electrochemical deoxyribonucleic acid biosensor based on carboxyl functionalized graphene oxide and poly-L-lysine modified electrode for the detection of tlh gene sequence related to vibrio parahaemolyticus

A carboxyl functionalized graphene oxide (GO-COOH) and electropolymerized ploy-L-lysine (PLLy) modified glassy carbon electrode (GCE) was fabricated and used for the construction of an electrochemical deoxyribonucleic acid (DNA) biosensor. The NH2 modified probe ssDNA sequences were immobilized on the surface of GO-COOH/PLLy/GCE by covalent linking with the formation of amide bonds, which was stable and furthur hybridized with the target ssDNA sequence. Differential pulse voltammetry (DPV) was used to monitor the hybridization events with methylene blue as electrochemical indicator, which gave a sensitive reduction peak at -0.287 V (vs. SCE). Under the optimal conditions the reduction peak current was proportional to the concentration of tlh gene sequence in the range from 1.0 x 10(-12) to 1.0 x 10(-6) mol L-1 with a detection limit as 1.69 x 10(-13) mol L-1 (3 sigma). The polymerase chain reaction products of tlh gene from oyster samples were detected with satisfactory results, indicating the potential application of this electrochemical DNA sensor. (c) 2012 Elsevier B.V. All rights reserved.