Herein, we report a facile multiplex enzyme-free electrochemical immunoassay for the detection of foodborne pathogens such as Listeria monocytogenes (Lm) using thionine labeled anti-Lm and Enterobacter cloacae (Ec) using ferrocene labeled anti-Ec as redox probes with Au nanoparticles- (NPs) decorated multiwalled carbon nanotubes (AuNPs-MWCNTs) composites as a detection platform. These nanocomposites have been characterized by Raman spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy dispersive X-ray analysis (EDAX) techniques. The individual linear range for Lm and Ec detection has been estimated to vary from 10^1-10^11 and 10^1-10^12 colony forming units (CFU)/mL, respectively. Conversely, the limit of detection (LOD) for Lm and Ec has been estimated to be 3.98 and 5.39 CFU/mL, respectively. Similarly, the multiplex detection of Lm and Ec showed a linear range of 10^1-10^7 (Lm) and 10^1-10^6 CFU/mL (Ec) with the LOD of 3.22 (Lm) and 4.17 CFU/mL (Ec). We have also demonstrated the application of our sandwich electrochemical immunoassay for the detection of Lm in Caenorhabditis elegans and Ec in milk and juice samples. © 2022 Elsevier B.V., All rights reserved.