A novel method was employed to directly and hom ogeneously attaching cupric oxide nanoparticles (CuONPs) on carbon nanofibers (CNFs) for sensitive amperometric nonenzymatic sensing of glucose. The obtained CuONPs-CNFs nanocomposite was characterized by transmission electron microscopy and X-ray diffraction. The CuONPs-CNFs nanocomposite modified glassy carbon electrode showed high electrocatalytic activity toward the oxidation of glucose in alkaline media and a nonenzymatic glucose sensor was constructed based on the functional nanocomposite modified electrode. Under optimal experimental conditions, the designed sensor exhibited a wide linear response to glucose ranging from 5.0 x 10(-7) to 1.1 X 10(-2) M with a high sensitivity of 2739 mu A mM(-1) cm(-2) and a low detection limit down to 0.2 mu M at the signal to noise ratio of 3. This sensor showed good accuracy, acceptable precision and reproducibility. Moreover, the proposed sensor was successfully applied in the detection of glucose in human blood serum indicating its possibility in practical application. (C) 2013 Elsevier Ltd. All rights reserved.