Staphylococcus causes disease conditions in poultry and widely distributed. Broilers may suffer from Staphylococcus aureus infection and its enterotoxins resulting in a major public health hazard and economic losses. This study aimed to detect the presence of Staphylococcus aureus producing Enterotoxin A in some broiler farms. Samples were collected from 200 broiler chickens, 60 apparently healthy (nasal swabs, tracheal swabs & cloacal swabs), and 140 diseased broilers (lung, liver, heart blood, spleen & thigh bone marrow).   Samples were collected aseptically from 10 randomly selected broiler farms in Bohera Governorate, Egypt, and were subjected for bacteriological examination for isolation and identification of S. aureus. The results of isolation of Staphylococcus from broiler chicken samples revealed that out of 200 bacteriologically examined broilers obtained from a total of 98 (49%) Staphylococcus isolates were isolated, 23  (38.3%) from apparently healthy and 75 (53.6%) from diseased birds. Also,  out of the  98 Staphylococcus isolates 20 (20.4%) were identified as coagulase positive S. aureus of which 4 isolates (17.4%) isolated from apparently healthy birds and 16 isolates (21.3%) from diseased ones. In apparently healthy birds, S. aureus was isolated from nasal swab, tracheal swab and cloacal swab samples with percentages of 10%, 12.5% and 40%, respectively. In diseased birds, S. aureus was isolated from lung, liver, heart blood, spleen and thigh bone marrow samples with  percentages of 15%, 24%, 20%, 20% and 26.7%, respectively. Antimicrobial resistance against 10 antimicrobial agents was performed and revealed that ciprofloxacin was the most effective antibiotic with susceptibility percentage of 95% followed by levofloxacin with sensitivity percent 75% and enrofloxacin, gentamycin (70% for each). All the S. aureus isolates were completely resistance against trimethoprim-sulfamethoxazole (100% resistance), followed by streptomycin, tetracycline and erythromycin with resistant percentages 80% for each. Variable degrees of sensitivity were reported for colistin sulfate and penicillin with  percentages of  55 % and 45%, respectively. Molecular identification of S. aureus Enterotoxin A (SEA) gene by PCR using specific primers revealed that 3 isolates out of 10 (30%) were positive for Enterotoxin A (SEA) gene. Our study concluded that broilers may act as a source of S. aureus which causes diseased problems leading to large economic losses, and broilers may be a source of (SEA) gene which can transmit to human causing illness.