This study was conducted to evaluate the rate of urinary tract infection at Assiut University Hospitals, and to determine the prevalence of extended-spectrum β-lactamase enzymes producing community- and hospital-acquired E. coli uropathogens at Urology Department in Assiut University Hospitals. Also, to perform the antimicrobial sensitivity testing for E. coli isolates to extended-spectrum cephalosporins according to CLSI. Phenotypic confirmatory tests (combined disc method, double disc diffusion method and ESBL-E-Test) were performed to test E. coli isolates for ESBL production and it was concluded that 30 (39.47%) of community isolates and 42 (70%) of nosocomial isolates were ESBLs producers.