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Abstract Background Beta-lactamase production remains the most contributing factor to beta-lactam resistance. Extended-Spectrum Beta-Lactamase-Producing Enterobacterales (ESBL-PE) are associated with risk factors both in hospital and community settings. Objectives To assess the incidence and risk factors for intestinal carriage of ESBL-PE among patients admitted to orthopedic ward of Mulago National Referral Hospital, and to analyze the acquisition of ESBL-PE during hospital stay and associated factors. Methods We screened 172 patients aged 18 years old and above who got admitted to the orthopedic ward of Mulago National Referral Hospital between May to July 2017. Stool samples or rectal swabs were collected at admission, every 3 days until fourteen days and screened for ESBL-PE. Data on demographic status, antibiotic use, admission and travel, length of hospital stay, hygiene practices and drinking boiled water were analyzed by logistic regression and cox regression model. Results At admission, 61% of patients showed intestinal ESBL-PE carriage. Co- resistance was common but no Carbapenem resistance was detected. Of the ESBL-PE negative, 49% were colonized during hospitalization. On admission, prior antibiotic use was significantly associated with carriage, but none was associated with acquisition during hospitalization at p-value