Background: Extended-spectrum beta-lactamases (ESBLs) are rapidly evolving group of beta-lactamase enzymes commonly produced by the Gram negative bacteria. Objectives: This work aimed to detect the bacterial pathogens that cause urinary tract infection and to detect ESBL producing bacteria among them in Pediatric Department at Benha University Hospital. Methodology: This study was conducted on 118 children patients suffering from UTI collected from Outpatient and Inpatient of pediatric department at Benha University during the period from January 2015 to the end of July 2015. They were 77 girls and 41 boys and their ages ranged from 1 to 13 years.118 Urine samples were subjected to isolation and identification of causative organisms, screened for ESBL production by the pattern of antimicrobial susceptibilities. Phenotypic identification for ESBL production was confirmed by double disc diffusion test, bla SHV and bla CTX-M genes in ESBL, producing isolates were detected by using multiplex PCR. Results: This study showed that the most common cause of children UTI was E.coli (55.1%) followed by klebsiella pneumoniae (21.2%). Thirty seven (31.4%) of 118 bacterial isolates that tested by double disc confirmatory test were ESBL positive. Among the ESBL producing bacteria, 16 out of 37 (43.24%) have CTX-m and SHV genes together while 3 out of 37 (8.1%) have SHV gene only. Conclusion: Our findings showed that majority of ESBL isolates were positive for both CTXm and SHV genes (43.24%) but SHV gene alone was positive only in (8.1%) of isolates, So studies should be done in different regions of Egypt to find the common ESBL genes present in that geographical area for epidemiological purposes. Also ESBL producing isolates have been increasingly recognized in hospitals in Egypt and are associated with multiple drug resistance. Thus, ESBL producing organisms should be identified quickly so that appropriate antibiotic usage and infection control measures can be implemented.