A new electrochemical immunosensor for serum myeloperoxidase (MPO) has been developed based on the self-assembly multilays of multi-walled carbon nanotubes (MWNTs), thionine (THI), gold nanoparticles (GNPs) and chitosanon (CHIT) on the glassy carbon electrode (GCE). The antibody of MPO (anti-MPO) was absorbed on the surface of GNPs monolayer. Horseradish peroxidase (HRP) was employed to block non-specific binding and amplify the response current signal. It was observed that the peak current was linear with the MPO concentration in a range of 2.5-125 µgl-1. The detection limit was 1.425 µgl-1 (S/N=3). Correlation analysis showed that this new immunosensor assay has a significant correlation with enzyme-linked immunosorbent assay (ELISA) (r=0.96, p>0.05) for 40 clinical specimens.