UTI is characterized by the evidence of uropathogens and pyuria and is accompanied by various clinical manifestations depending on the area of involvement. The aim of this study - was to isolate the bacterial infection to urinary tract infection in men and women at different ages and to investigate the  susceptibility of bacteria implicated in urinary tract infection to different antimicrobial drugs (antibiotics).Also to determine minimum inhibitor concentration for resistant bacteria to antibiotic and  molecular characterization of S. aureus  associated  . Materials and methods :One hundred  urine specimens were received from patients admitted during January till December 2015. S. aureus isolates were confirmed by typical colonial morphology, and identified by differential tests as well as by the growth  on characteristic agar, Susceptibility testing was carried out by disk diffusion method . Among the 100 isolates, 6 bacterial strain  was isolated from the urine  specimens of infected Patient admitted at Port-said area out-patient clinics, using agar medium and macCkonkey agar, This organism was characterized by  biochemical tests and showed similarity with S. aureus . The genomic level confirmation done with 16S rDNA primer by submitting the genomic sequence to Gene Bank under acc.No-GU046545 after comparing, showed  98 % sequence similarity with S. aureus  . Antibiotic susceptibility test revealed that Amikacin, Amoxicillin and Imepenem are the lowest resistant rate with percent of 100, 83 and 66%respectively and Ciprofloxacin showed the highest resistant rate with 84% .  Pattern on antibiotic susceptibility test showed high resistant rate to some antibiotic that made it difficult for pregnant patients, although its frequency was low. The most prevalent Gram positive bacterial pathogen in Port-saed city was S. aureus and the most sensitive antibiotic against it is Amikacin.