Background . The need to reduce the transmission of infectious diseases makes the use of personal protective equipment and safety medical devices compulsory among hospital laundry staff. The practice, however, remains to be low among hospital laundry staff members. Globally, not many studies seem to have been carried out to sufficiently tell us about the barriers to personal protective equipment use among hospital laundry workers. Related studies in Ethiopia are even fewer. This study assessed the barriers to personal protective equipment use among laundry staff of government hospitals in Hawassa City, Southern Ethiopia, 2019. Methods . Two qualitative data-gathering methods—focus group discussions and key informant interviews—were used to collect data for this study. Eight focus group discussions were conducted with hospital laundry workers. Similarly, six key informant interviews were held with Infection Prevention and Patient Safety Officers. Thematic analysis was performed using Open Code 4.02. Result . Organizational- and individual-level barriers such as unavailability of essential personal protective equipment, a disharmonious work environment, low perception of susceptibility, and belief about personal protective equipment interference with work performance were identified as the major barriers to personal protective equipment use in the present study. Conclusion . Organizational- and individual-level barriers have been identified as causes for the low level of personal protective equipment use among hospital laundry workers. Therefore, improving institutional supplies in quantity and quality may have a positive implication for the improvement of infection prevention practices in the study area. Also, designing sustainable strategies and raising laundry workers’ awareness of a safe work environment may lead to the improvement of infection prevention practices.