Escherichia coli account approximately to 85% of the Urinary tract infection. UTI affect the different parts of the urinary tract and is considered as a common bacterial infection. The infection is caused as a consequence of the urinary tract bacterial invasion. E. coli were isolated from the urine of UTI patients referred to King Abdulaziz Specialist Hospital, Taif, Saudi Arabia. Antibiotics susceptibility was tested on 25 antibiotics using the disc diffusion method. The prevalence of antibiotics resistance genes was realized by Polymerase Chain Reaction (PCR). Results showed heterogenicity in the percentage of antibiotics resistance from 100% for penicillin to 2% for imipenem. 30% of the isolates appeared as for Extended-Spectrum Beta-Lactamase (ESBL) positive and 74% are multidrug resistance strains. Distribution of antibiotic resistance genes showed that aac(3)-IV and blaSHV genes were identified in 33.33% of isolates. In addition, qnrA, blaCMY and dfrA1 genes were founded in 37.25%, 19.60% and 17.64% of the isolates respectively. In total, 17 different genotypes were detected, and 12 isolates (24%) do not include any genes in their genomes. Multi-drug resistant E. coli have antibiotics profiles highly variable and the mechanism of resistance was not correlated to the investigated genes.