Multi-walled carbon nanotubes (MWNTs) were employed as the rigid template for the assembly of gold/copper alloy nanoparticles (GCNPs). The MWNT/copper composite film was codeposited on glassy carbon electrode at a constant current and subsequently immersed in aqueous chloroauric acid. The GCNPs, prepared by a simple galvanic reaction, are strongly bound to the MWNTs to form a uniform and compact film on glassy carbon electrode (GCE). Cyclic voltammetry and scanning electron microscopy were used to characterize GCNP/MWNTs. The oxidation process of glucose at the GCNP/MWNT shows two well-defined peaks at -0.30 V and 0.023 V in alkaline aqueous solution. The experimental results indicate that the modification of GCNP/MWNT on the surface of GCE significantly improved the electrocatalytic activity towards the oxidation of glucose.