Introduction: Extended Spectrum Beta-lactamases (ESBLs) are responsible for multi-drug resistant strains of Escherichia coli and Klebsiella species worldwide. They are not routinely tested for and may be responsible for clinical failure in treatment of urinary tract infection in University College Hospital, Ibadan. Aim: The aim of this study was to evaluate the prevalence of ESBL-producing Escherichia coli and Klebsiella species in urinary tract infection in University College Hospital, Ibadan and identify the associated factors. Methods: In this cross-sectional survey, Escherichia coli and Klebsiella species causing urinary tract infection at the University College Hospital were studied with corresponding clinical details obtained from clinical notes. Associated factors were determined using correlation and multivariate analyses.