We describe a central venous catheter-related (Port-A-Cath; Smiths Industries Medical Systems [SIMS] Deltec, Inc., St. Paul, Minn.) infection caused by Rhodotorula glutinis in a 51-year-old man with nasopharyngeal carcinoma. He was treated with fluconazole for 8 weeks and had the catheter removed. Two isolates of R. glutinis recovered from blood specimens (one obtained via peripheral veins and one via the catheter) before administration of fluconazole and one recovered from the removed catheter 17 days after initiation of fluconazole therapy exhibited high-level resistance to fluconazole (MICs, >256 microg/ml). These three isolates were found to belong to a single clone on the basis of identical antibiotypes determined by the E test (PDM Epsilometer; AB Biodisk, Solna, Sweden) and biotypes determined by API ID32 C (bioMerieux, Marcy I'Etoile, France) and their identical random amplified polymorphic DNA patterns.