To determine the prevalence and the antibiotic resistant patterns of the multi-drug resistant Extended-Spectrum Β-Lactamase(ESBL) producing E. coli isolates from children urine samples, in Queen Rania Al-Abdullah Hospital for children.A total of 61 non-repetitive urine samples from various outpatient clinics and inpatient wards were collected retrospectively over a period of 5 months (May 2012 to September 2012). The resistant patterns, screening and confirmatory tests for phenotypic detection of ESBL-producers were studied using the VITEK 2 system against a set of antibiotics found on the antimicrobial susceptibility extend card AST-EXN8.Children were nearly equally infected by both types of E. coli isolates, ESBL-producers 31 (50.8%) and non ESBL-producers 30 (49.2%). ESBL-producing E. coli showed maximum rate resistance to Cefuroxime and Piperacillin (100%), Aztreonam, Cefixime, Ceftriaxone plus Levofloxacin (96.8%), Ampicillin/Sulbactam and Cefepime (93.5%), and Moxifloxacin (90.3%), while minimum resistance rate was seen with Tigecycline (12.9%), Colistin (3.2%) and meropenem (0%). ESBL-producing isolates were significantly more resistant than Non-ESBL-producers (p < 0.05) to the following antimicrobials (Ampicillin/Sulbactam, Aztreonam, Cefepime, Cefixime, Ceftriaxone, Levofloxacin, Moxifloxacin, Piperacillin and Tetracycline). Multi-drug resistance was found to be higher in ESBL-producing isolates, which were resistant to at least 9 antibiotics. To limit the spread of the multi-drug resistant ESBL-producers E. coli isolates, we should perform screening test for these isolates on daily basis, isolate the infected patients and choose the best therapeutic option.According to the resistant pattern and safety issue, Morepenem can be considered as first line treatment and colistin as last resort therapy.