Food is the primary cause for diseases in humans and carries high risk pathogens. Assessment of the safety in foods is needed to validate the presence of pathogenic bacteria. We used colony PCR for this approach to detect foodborne pathogens such as Escherichia coli, Lactobacillus and Bacillus cereus. Suitable primers were selected based on specific gene 1040 for Escherichia coli, gene S2 for Lactobacillus, and gene NVF for Bacillus cereus. Agarose gel electrophoresis is used for the detection of amplified products against a suitable marker. ImageJ is used for DNA band analysis, enabling precise quantification, normalization, and statistical comparisons. These studies have established a promising role in the detection of pathogens in various environmental samples. The insights gained from this study may serve as the foundation for rapid detection of foodborne diseases in the food industry.