Spinel-type MnCo2O4 nanofibers (MCFs) were successfully synthesized by electrospinning and sequential calcination. The crystal structure, composition and morphology of the synthesized MCFs were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, energy-dispersive spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy and transmission electron microscopy. Due to the outstanding electron-transfer ability of spinel MnCo2O4 and the large surface area of nanofibers, the synthesized MCFs were employed as electrocatalysts for the oxidation of glucose. Cyclic voltammetry and amperometry were used to evaluate the electrocatalytic activities of the MCFs towards glucose. The non-enzymatic glucose sensor showed a wide linear range of 0.05-800 mu M with a low detection limit of 0.01 mu M (S/N = 3).