Background: The inappropriate use of antibiotics is a global threat. Clinical microbiology laboratory testing can support optimal antibiotic prescribing for many conditions. The purpose of this study was to characterize antibiotic usage in the context of treatment of uncomplicated urinary tract infection (UTI) received through a Ghanaian Accident & Emergency, which found high rates of improper usage. Methods: A prospective cohort study recruited patients >18 years of age who were admitted to a large Ghanaian teaching hospital with suspected UTI. Eligible patients were identified through a daily review of admission logs. Data were collected through a review of medical records and microbiology laboratory urine data. Results: A total of 90 patients were enrolled, but urine cultures were obtained from only 50 (56%) patients. Most positive urine cultures grew typical urinary pathogens (21 of 25, 84%). The most common empiric regimens were cephalosporin-based and did not vary if additional infections were suspected or a urine culture was not sent. The majority of patients with confirmed UTI had isolates that were not susceptible to antibiotics selected for empiric treatment (18 of 21, 86%). Although more than half had their empiric regimen changed between admission and study follow up, only 42% (5 of 12) were switched to a regimen that included agent(s) to which their urinary isolate was known to be susceptible. Conclusions: Establishing hospital-wide guidelines regarding the evaluation and treatment of patients with suspected UTI may help improve antibiotic utilization and patient outcomes by increasing the use of urine cultures and tailoring therapy in response to culture results.