All methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from colonized or infected patients in a 625-bed public teaching hospital during an epidemic, and for 3 years thereafter, underwent susceptibility testing to mupirocin. Mupirocin resistance among MRSA increased markedly over this period (1990, 2.7%; 1991, 8.0%; 1992, 61.5%; 1993, 65%) in association with increased use of mupirocin ointment as an adjunct to infection control measures.