There is still a significant outbreak of diseases linked to foodborne pathogens such as bacterial, fungi despite safety measures, and rigorous food preservation regime by authorities in charge. These pathogens constitute a significant risk to overall public health. Early detection of these pathogenic microbes is, therefore, essential for food safety and to mitigate the health risks that they pose. Most of the detection techniques used include biochemical techniques, culture-based techniques, biosensors, and methods based on spectroscopy, immunology, and molecular-based methods (polymerase chain reaction and sequencing). For this chapter, we focused on the biochemical techniques used to analyze or detect gram-positive toxin producing microorganisms.