The presence of enterotoxigenic Staphylococcus and its enterotoxins in milk and dairy products is a concern for industry and government agencies responsible for public health. S. aureus is the species most often associated with outbreaks or cases of staphylococcal food poisoning. As this microorganism is an important pathogen of mastitis in cattle, the udder of the infected animal is one of the main sources of contamination of raw milk. Other species of Staphylococcus have also been associated, but with much less frequency with staphylococcal toxinosis of food origin. This study aimed to evaluate the enterotoxigenic potential of 300 Staphylococcus spp. isolated from the mammary gland of cows from herds in the State of Minas Gerais, from pasteurized milk and Minas frescal cheeses sold in Juiz de Fora, MG and carry out the molecular characterization of enterotoxigenic strains. Strains with enterotoxigenic potential were identified using the PCR technique by amplifying fragments of the genes encoding the classic SEA-SEE enterotoxins (i.e. sea, seb, sec, sed, and see) and the new SEG-SEL enterotoxins (i.e. sec, seh, know, sej and sel). The isolates were analyzed by PCR for the femA gene to identify S. aureus. For the other isolates, only those confirmed to be enterotoxigenic by PCR, were identified through the API®-STAPH system (BioMérieux, Marcy-l`Etoile - France). The potentially enterotoxigenic isolates had their electrophoretic profile determined by pulsed field electrophoresis (PFGE) and sequenced gene fragments of enterotoxins. This study contributed to a better understanding of the enterotoxigenic Staphylococcus species, the enterotoxin genes and the predominant clones.