The present study was carried out to spotlight on the presence of Staphylococcus aureus (Staph aureus) among rabbit farm and to characterize the bacterial isolates for the presence of blaZ, coa, mecA, tetK and nuc gene, It also aimed to demonstrate the antimicrobial susceptibility of S. aureus in vitro and detect its pathogenicity in weaned rabbits. A total of 426 samples were collected included 98, 257 and 71 samples from apparently healthy rabbit, diseased rabbits and freshly dead rabbits. Staph aureus was isolated in 20(20.4%) out of 98 apparently healthy rabbits, 78(30.4%) out of 257 diseased rabbit and 22(31%) out of 71 dead rabbits and its presence was confirmed through cultural and biochemical characterization. Multiplex PCR was performed on the isolated strains of S. aureus which showed the presence of blaZ, coa, mecA, tetK and nuc virulent genes. All isolates were positive for the gene coa, mecA, tetK and nuc virulent genes. This results indicates that the coagulase-positive of S. aureus was the most predominant cause of skin disease in rabbits disease and carried the mecA and nuc virulent gene. Antibiotic sensitivity against conventional antimicrobials revealed that Staphylococcus aureus strains were sensitive to streptomycine (70%) and penicillin, gentamycin   (63.3%) in addition to doxycycline (60%) and resistance to ciprofloxacin and Amoxicillin (75%). As well, high degree of resistance to Trimethoprim /Sulphathoxazol, Cefotaxime, Cephradin (100%) and Colstine sulphate were detected 72.3%. The pathogenicity test conducted on of 5 week old weaned rabbits showed that S. aureus was highly pathogenic resulting in mortality rates of 65 and 35%, respectively. The rabbits exhibited septicaemia with congestion and peticheal hemorrhage throughout the intestine, sever congestion of the liver, spleen and kidneys,as well as the congestion of the intestine blood vessels and lung. Pneumonia or abscesses were also observed in the lungs or heart. Histopathological changes supported the so observed lesions. In conclusion, the detection of S. aurues virulent genes with overlapping functions and the reproduction of antibiotic resistance genes in the Egyptian field is very importance additionally, in vivo pathogenicity studies. S. aureus was identified as the most significant pathogen causing death of rabbits in rabbit farms. Therefore, accurate, rapid and molecular diagnostic methods of must be implemented. Finally, it is essential to conduct in vitro sensitivity tests of S. aurues to different antimicrobial agents due to the observed resistance to the commonly used antibiotics for prevention and control S. aureus in rabbits.