•Ni-Co NWAs electrode was fabricated by polycarbonate template.•Ni-Co NWAs electrode with 10% of Co molar ratio shows best catalytic activity.•Direct urea/H2O2 fuel cell shows high output performance with Ni-Co NWAs anode. Nickel-cobalt nanowire arrays (Ni-Co NWAs) electrode is prepared by one-step galvanostatic electrodeposition with a polycarbonate membrane as the template. By adjusting the Co proportion in the Ni and Co bath solution into 10%, the optimal Ni-Co NWAs electrode in terms of relatively lower onset potential and highest current density towards urea electro-oxidation is obtained. Its catalytic performance is investigated by constructing single direct urea/hydrogen peroxide (H2O2) fuel cell. Results show that a peak power density of 7.4mWcm−2 and an open circuit voltage of 0.92V are achieved at room temperature when 9.0molL−1 KOH and 0.33molL−1 urea are used as the anolyte, H2SO4 and H2O2 as the catholyte. Additionally, the urea/H2O2 fuel cell also demonstrates excellent stability during short term duration test.