BACKGROUND Endoscope-associated infections are reported despite following proper reprocessing methods. Microbiological testing can confirm the adequacy of endoscope reprocessing. Multiple controversies related to the method and interpretation of microbiological testing cultures have arisen that make their routine performance a complex target. OBJECTIVE We conducted a pilot study using disposable bronchoscopes (DBs) to simulate different reprocessing times and soaking times and to compare high-level disinfection versus ethylene oxide sterilization. We also reviewed the time to reprocessing and duration of the procedures. METHODS Bronchoscopes were chosen because an alternative disposable scope is commercially available and because bronchoscopes are more prone to delays in processing. Disposable bronchoscopes were contaminated using a liquid bacterial suspension and were then incubated for 1-4 hours. Standard processing and high-level disinfection were performed on 36 endoscopes. Ethylene oxide sterilization was performed on 21 endoscopes. Endoscope cultures were performed using the standard brush, flush, brush technique. RESULTS After brushing was performed, a final water-flush culture procedure was the most effective method of detecting bacterial persistence on the disposable scopes. Klebsiella pneumoniae was the most commonly recovered organism after reprocessing. Ethylene oxide sterilization did not result in total elimination of viable bacteria. CONCLUSION Routine endoscopy cultures may be required to assess the adequacy of endoscopic processing.