One of the most important pathogens that threaten human health all over the world is Staphylococcus spp. characterization and identification of such pathogen considers a useful tool to control of some serious problems resulting by these bacteria. Therefore one hundred and twenty samples including blood, urine, abscess, semen, pus, sputum, ears, vaginal and spit swabs were collected from patients of Tanta University Hospital and outpatient clinics. A total of 126 Gram positive bacterial isolates were obtained from these clinical specimens. Of these isolates 107 bacterial strains were identified as Staphylococcus spp and 15 strains identified as Streptococcus spp in addition to 4 strains were identified as Enterococcus spp, preliminary identification was conducted on the basis of colony characteristics such as Gram staining, pigment production, hemolysis, catalase activity, coagulase test and fermentation of manitol sugar. Out of these strains, 30.95% have the potency to make alpha hemolysis while 30.95% have the ability to make beta hemolysis and 38.09% posse the capacity to make gamma hemolysis on blood agar medium. Beta hemolysis Staphylococcus spp were selected for study of some virulence factors on basis of coagulase production in which 17.24% of Staphylococcus spp  were coagulase positive and 82.76% were coagulase negative Staphylococcus spp. Studying the susceptibility pattern of these strains to some commercial antibiotics was carried out. Further future studies are recommended to investigate the effect of some natural compounds on gene regulation that responsible for hemolysis process.