This study investigated the prevalence, risk factors and antimicrobial resistance of Staphylococci and E. coli isolated from diabetic urinary tract infection patients. Urine samples collected from 694 cases admitted to the General Health Insurance Authority Hospital in Port Said City, Egypt.  A total of 302 bacterial strains were isolated from 292 urine samples with glucosuria and infection positive. The prevalence of the S. saprophyticus, S. aureus, E. coli and other species isolates from the urine samples are 54.7%, 34%, 7.6% and 3.7%, respectively. The antibacterial resistance profile of the isolated bacteria was investigated against different antibiotics. Most bacterial isolates were more susceptible to oflaxacin antibiotic while resistance to ampicillin. Multiple antibiotic resistance (MAR) was found highest for bacterial isolates obtained from urine samples. S. aureus showed high resistance percentage than other bacterial strains to studied antibiotics. Vancomycin resistance was detected in 23% of all S. aureus isolates. Out of 165 isolates S. saprophyticus, 71% were ג-lactamase producers, while all S. aureus and E. coli isolates were ג-lactamase positive. Also the purpose of this study was to study any relative between the recognized antibiotic resistance of bacteria and the occurrence of plasmids. Molecular sizes of the detected plasmids were 39.306 kbp in S. aureus and 44.640 kbp in E. coli. Plasmid curing in combination with MIC determination revealed that resistance of staphylococci and E. coli isolates was plasmid-mediated linked. The high MAR recognized makes it essential for antibiotic resistance testing to be accompanied prior to antibiotics remedy for diabetic urinary tract infection patients.