The present work demonstrates fabrication of a non-enzymatic glucose sensor based on CuO nanoparticles deposited over poly(3,4-ethylenedioxythiophene) (PEDOT) conducting polymer infiltrated with MoS2. Structural, morphological and elemental analysis of the fabricated sensor electrodes were performed via different characterization techniques like x-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), elemental dispersive x-ray spectroscopy (EDX), and Fourier transform infra-red spectroscopy (FTIR). The cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) studies of the hybrid nanoelectrode (CuO/PEDOT-MoS2) exhibited better redox activity and electron transfer kinetics, as compared with the CuO/PEDOT and CuO only systems. Moreover, the electrochemical parameters of all the systems were determined and compared accordingly while CuO/PEDOT-MoS2 hybrid system offers a significant enhancement in the electroactive area (~1.47 cm-
2) and rate constant (0.76 s-1) towards oxidizing glucose into gluconic acid. In the CV responses, an augmented activity was monitored at +0.6 V which was considered as the dc bias potential in the chronoamperometric experiment for detecting glucose. The sensor electrode yielded a low LOD of 0.046 μM and with a sensitivity magnitude as high as 829 µA mM-1 cm-2 over a wide linear range, between 30 μM to 1.06 mM of glucose concentration.