Highly selective and efficient nonenzymatic electrochemical glucose sensors were fabricated by using electrospun polyvinylidenefluoride-co-hexafluoropropylene (PVdF-HFP) composite nanofiber membranes. The homogeneous, smooth and strongly interconnected nanofibers were identified for the bare PVdF-HFP membrane and beads in the PVdF-HFP nanofibers were observed for the PVdF-HFP/Ni and PVdF-HFP/Ni/Co nanofiber membranes. The face-centered cubic structure of Ni nanoparticles doped in the PVdF-HFP nanofibers has not been altered, even after the alloy formation with Co. The electrochemical oxidation of glucose in an alkaline medium was chosen as a probe for the detection of glucose and was achieved through the fabricated nanofiber membranes. Among the fabricated nanofiber membranes, the PVdF-HFP/Ni/Co membrane exhibited an excellent sensing behavior toward glucose with the low limit of detection and linear range of 0.26 mu M and 1 mu M to 7 mM, respectively. Furthermore, the fabricated nanofiber membrane exhibited good selectivity, high stability and reproducibility, which promises its applications in nonenzymatic glucose sensors.