We have produced working fiber-optic-based biosensors that have detected aflatoxin B1(AFB1) in foods which were previously assayed with standard methods. In this study, we compare fiber-optic biosensor detection levels with those of a conventional ELISA. These detections require little specialized training of technicians, and as little as one hour to perform. AFB1concentrations were measured by several fiber-optic approaches based on fluorescence. We detected AFB1by its native fluorescence, and alternatively, in the competition mode with either FITC-labelled antibody or enzyme-labelled antibody. Extractions of AFB1from food sources (peanut and corn meal) were made and successfully tested using fiber-optics. Detection of 0.05 ng/mL AFB1was found with both of these sources using the native fluorescent format.