Foodborne disease is one of the major public health problems worldwide. The conventional methods used to detect foodborne pathogens require multiple incubation steps and take a long time to results. Thus, there is an urgent need for biosensors that can determine pathogens without special sample preparation. This paper outlines a method for discriminating E. coli contaminated meat since this bacterium has been implicated as one of the main causative agents of food illness. The method is based on the laser-induced fluorescence response of the Flavin group that is presented among E. coli bacteria’s metabolites. The fluorescence activity of metabolites produced by five E. coli strains was investigated. The list of analyzed strains contains ATCC 25922 E. coli, Enteropathogenic E. coli, Enteroinvasive E. coli, Enterotoxigenic E. coli, and Enteroaggregative E. coli. The preliminary results have allowed developing the statistical model using a part of the fluorescence spectra in the range of 520-560 nm when excited at 450 nm. The proposed model is aimed to differentiate contaminated and uncontaminated meat samples. It has been confirmed that the examined technique provides detection of a bacterial concentration of 106 CFU∙cm-2 in five hours after initial contamination at room temperature. The further improvement of the method by using fluorescent probes is discussed.