Screen-printed copper oxide (CuO) and CuO/few-layer graphene on graphite electrodes were used to fabricate the ultrasensitive nonenzymatic glucose biosensors. Flower-like CuO and flower-like CuO/few-layer graphene composites were prepared by screen-printing method and characterized by X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), and high-resolution transmission electron microscopy (HETEM). On the basis of their cyclic voltammetry (CV) and chronoamperometry results, it was concluded that the addition of graphene to CuO significantly improved the performance of the fabricated glucose sensors, exhibiting high and reproducible sensitivity of 3120 mu AM(-1) cm(-2) with three linear ranges from 4 M to 13.5 mM and the detection limit of 4 mu M (S/N = 3) in a fast response time of 2 s. In addition, the fabricated sensors could effectively avoid the disturbance by interferents, such as Ascorbic Acid (AA), Uric Acid (UA), and Dopamine (DA). Most importantly, the testing results of real blood serum samples demonstrated that the electrodes were applicable and acceptable for the determination of glucose concentrations in human serum. The efficiencies of two non-enzymatic glucose biosensors for glucose determination were comparable with that of a commercial enzymatic sensor. (C) 2016 Elsevier B.V. All rights reserved.