S. aureus produces extracellular protein toxins and virulence factors which contribute to the pathogenicity of the organism. The present work aimed to analysis the prevalence of enterotoxin producing S. aureus strains isolated from bovine and humans origin using reverse passive latex agglutination (RPLA) kit and PCR. A total of 25 S. aureus isolates out of 374 samples collected from cattle, buffaloes and human were identified, 20 from bovine and 5 from human. Using RPLA, 10 out of the 25 S. aureus isolates were found to be toxigenic with an incidence of 40%. They were distributed as enterotoxin C (50%), a (20%), A& B, A& C and enterotoxins A, B, C and D (10 % each). Using PCR 7 enterotoxigenic S. aureus isolates (28%) were detected. sea gene was detected in 2 isolates (28.6%), while sec, seb, sea & seb, sec & see and sea, sec, sed genes were detected in one isolate each (14.3% each). Analysis of the results obtained by RPLA and PCR for the productivity of classical enterotoxins A-D revealed approximately correlation between each other. It could be concluded that cows' and buffaloes' milk are of public health risk due to potent staphylococcal food poisoning strains.