Nucleocapsid-like Au@NiO@CuO nanocomposites (NCs) with a size of similar to 164 nm were synthesized by via a simple situ precipitation. Field scanning electron microscopy (FESEM), X-ray powder diffraction (XRD) and Raman were employed to characterize the morphologies and analyse the composition of the as-prepared samples. And according to electrochemical studies, MWCNT, as a carrier, showed good chemical stability and excellent electronic conductivity. Therefore, in our work, differential pulse voltammetry (DPV) and cyclic voltammetry (CV) demonstrated that the synthesized nanocomposites were applied to the electrochemical determination of glucose owing to its capability, such as good chemical stability and excellent electronic conductivity. The non-enzymatic glucose sensor displayed low detection limit of 0.15 mu M with linear range of 0.001-5.6 mM in 0.20 M NaOH as the solvent, suggesting that this sensor possessed excellent accuracy and stability. The good recoveries achieved in human serum samples indicated that the novel MWCNT-Au@NiO@CuO/GCE biosensor has a good application prospect in glucose detection device.