A sensitive electrochemical immunoassay for rapid detection of Escherichia coli has been developed by anodic stripping voltammetry (ASV) based on core-shell Cu@Au nanoparticles (NPs) as anti-E. coli antibody labels. The characteristics of Cu@Au NPs before and after binding with antibody were confirmed by transmission electron microscopy (TEM). After Cu@Au-labeled antibody reacted with the immobilized E. coli on Polystyrene (PS)-modified ITO chip, Cu@Au NPs were dissolved by oxidation to the metal ionic forms, and the released Cu(2+) ions were determined at GC/Nafion/Hg modified electrode by ASV. The utilization of GC/Nafion/Hg modified electrode could enhance the sensitivity for Cu(2+) detection with a concentration as low as 9.0 x 10(-12)mol/L. Since Cu@Au NPs labels were only present when antibody reacted with E. coli, the amount of Cu(2+) directly reflected the number of E. coli. The technique could detect E. coli with a detection limit of 30CFU/mL and the overall analysis could be completed in 2h. By introducing a pre-enrichment step, a concentration of 3CFU/10mL E. coli in surface water was detected by the electrochemical immunoassay.