In this work, novel NiO–CdO composite nanofibers with excellent conductivity and electrocatalytic property are reported. The hybrid nanofibers are fabricated by a facile two-step synthetic route consisting of electrospinning and subsequent calcination at 500 °C. EDX element mapping indicates the homogeneous distribution of CdO and NiO along the hybrid nanofiber. The incorporation of CdO into the NiO nanofibers is found to significantly suppress the grain size of the NiO crystallites. HRTEM images clearly reveal the dislocation areas with the displacement of lattice lines between CdO and NiO, which might contribute to the significantly improved properties of NiO nanofibers after CdO-doping. The as-prepared NiO–CdO nanofibers are further applied to the non-enzymatic glucose detection and their glucose response is around 9-fold and 1650-fold higher than those obtained with pristine NiO nanofibers and CdO nanofibers, respectively, in the cyclic voltammetry studies. The simple strategy of doping conductive CdO into a semi-conductive metal oxide with dislocation feature opens a new route to generate novel conductive metal oxide composite nanofibers with outstanding functions, which may have great potential applications ranging from sensors to energy area.