A novel glucose biosensor, based on the modification of well-aligned polypyrrole nanowires array (PPyNWA) with Pt nanoparticles (PtNPs) and subsequent surface adsorption of glucose oxidase (GOx), is described. The distinct differences in the electrochemical properties of PPyNWA-GOx, PPyNWA-PtNPs, and PPyNWA-PtNPs-GOx electrodes were revealed by cyclic voltammetry. In particular, the results obtained for PPyNWA-PtNPs-GOx biosensor showed evidence of direct electron transfer due mainly to modification with PtNPs. Optimum fabrication of the PPyNWA-PtNPs-GOx biosensor for both potentiometric and amperometric detection of glucose were achieved with 0.2M pyrrole, applied current density of 0.1mAcm-2, polymerization time of 600s, cyclic deposition of PtNPs from -200mV to 200mV, scan rate of 50mVs-1, and 20 cycles. A sensitivity of 40.5mV/decade and a linear range of 10μM to 1000μM (R2=0.9936) were achieved for potentiometric detection, while for amperometric detection a sensitivity of 34.7μAcm-2mM-1 at an applied potential of 700mV and a linear range of 0.1-9mM (R2=0.9977) were achieved. In terms of achievable detection limit, potentiometric detection achieved 5.6μM of glucose, while amperometric detection achieved 27.7μM.