An electrochemical DNA biosensor was fabricated by integrating probe with novel chitosan(CH)-V0.5Ti0.5O2(VTO) nanocomposite onto indium-tin-oxide (ITO) coated glass plate.The results reveal the presence of V0.5Ti0.5O2 (VTO) can enhance electroactive surface area of the CH-V0.5Ti0.5O2 nano-biocomposite that provides desirable environment for loading of DNA with better conformation leading to increased electron transfer kinetics between the medium and ITO electrode.The properties of modified electrodes were studied by electrochemical impedance spectroscopy (EIS) using [Fe(CN)6]3?/4? as the redox indicator.Differential pulse voltammetric (DPV) studies have been conducted to monitor the hybridization events using methylene blue (MB) as a hybridization redox indicator.The observed results show promising application of the CH-V0.5Ti0.5O2 nanocomposite for the detection of other biological and chemical molecules.