Target label-free, reagentless electrochemical DNA biosensor based on sub-optimum displacement

One of the most time consuming and complex steps in the detection of DNA target with a biosensor is the previous labeling of the target. In this paper, a novel target label-free, reagentless and easy to use DNA biosensor is reported. Electrochemical transduction (cyclic voltammetry, differential pulse voltammetry and impedance spectroscopy) and optical red out by surface plasmon resonance were chosen for the platform optimization. This target label-free DNA detection method is based on displacement of sub-optimum labeled oligonucleotide. This strategy requires the pre-hybridization of the capture probe immobilized on the electrode surface with a sub-optimum mutated oligonucleotide pre-labeled with an electrochemically active ferrocene moiety. Due to the higher affinity of the target that is fully complementary to the capture probe, the sub-optimum ferrocene-labeled sequence is displaced when the fully complementary target is introduced into the system. The decrease of the electrochemical signal from the ferrocene verifies the presence of the target, which is proportional to the target concentration. A variation of this strategy was employed to enhance the ferrocene signal. A diffusional mediator, ferrocyanide, was introduced in the system to help in this purpose. This platform attains a stable, specific and reproducible response (5-15%), with a detection limit in the range of microM. This electrochemical sensor is the first example of this kind of sensor to detect cystic fibrosis, however, this configuration could be generically applied to any application where the detection of a DNA target is involved.