In this study, a nano Ag-CuO/rGO (reduced graphene oxide) composite was designed and constructed as a novel nonenzymatic glucose sensor. The composite was fabricated through a one-step synthesis process. In the process, Ag-CuO with an average particle size of 10 nm formed and dispersed homogeneously on the surface of rGO sheets. When used for nonenzymatic glucose sensing, the resultant Ag-CuO/rGO composite showed a high sensitivity of 214.37 μA mM −1 cm −2 and an extremely wide linear response from 0.01 to 28 mM with a 0.76 μM detection limit (S/N = 3) at +0.6 V. The excellent sensing properties of the composite are probably due to the synergistic effect of the combination of Ag, CuO nanoparticles and rGO. The electron transfer is improved by the addition of Ag nanoparticles, and the composite electrode possesses larger surface area due to the rGO. The Ag-CuO/rGO composite doe not only show the good catalytic activity, excellent selectivity but also outstanding long term stability, good reproducibility, which makes it a novel type of composite for nonenzymatic glucose sensing.