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As the safety in the food supply becomes critical, the demand for rapid, low volume, sensitive and economic biosensor devices have dramatically increased. A novel immunosensor based on electrochemical sandwich immunoassay has been developed for pathogen detection in food. The immunosensor design is based primarily upon the specific nature of labeled antibodyantigen binding. The architecture of the biosensor demonstrates the advantages of using lateral flow format strip attached to a portable circuitry for signal measurement. The ability to change the specificity of the antibodies enables the biosensor to be used as a semi-quantitative detection device for many types of foodborne pathogens. Results show that the biosensor can detect a very low concentration of cells in less than 10 minutes. In addition, the biosensor gives a higher specificity and sensitivity than conventional approaches. Details on the device fabrication and performance of the biosensor in detecting pathogenic organisms in fresh produce are presented. Application of the device can be used to enhance food safety and biosecurity of the food supply chain, as well as it can be used for medical diagnostics and bio-defense measures.