This study reports a novel CuO nanoparticles (NPs) supported on S-doped graphene (CuO/SG) as an electrode material for nonenzymatic glucose detection. The CuO/SG was successfully synthesized by a facile microwave-assisted approach, and CuO/SG composed of ∼13nm of CuO NPs being relatively well-distributed and strongly anchored on SG was obtained. Electrochemical test revealed that the CuO/SG based electrode exhibited a higher electrocatalytic activity toward glucose oxidation than CuO NPs and CuO/reduced graphene oxide (rGO) modified electrodes owing to the synergistic effect between CuO NPs and SG. As a consequence, the CuO/SG based sensor exhibited a rapid response of 2s, a wide linear range of 0.1–10.5mM, a high sensitivity of 1298.6μA·mM−1 ·cm−2, and a low detection limit of 80nM, along with the excellent selectivity, high stability, and good accuracy in real sample analysis, thus leading to a promising candidate for nonenzymatic glucose detection.