The application of typing procedures for the purpose of strain differentiation among isolates of Candida albicans obtained from hospitalized patients has been limited. We have applied biotyping and DNA restriction fragment analysis (DNA fingerprinting) by using EcoRI to the study of C. albicans isolates obtained from hospitalized patients. A total of 68 isolates from 15 patients were studied. Thirteen subtypes were identified by biotyping, 8 by DNA fingerprinting, and 21 by a combination of the biotyping and DNA fingerprinting approaches (composite subtype). Both techniques were highly reproducible. In examining the strain variation among isolates obtained from multiple anatomic sites over time, we found that similar, if not identical, strains were recovered from the oropharynx, urine, stool, and blood in a given patient, and these strains persisted. Only rarely did two patients share the same composite subtype suggesting sporadic nosocomial transmission. The combination of biotyping and DNA fingerprinting improved strain discrimination compared to either method alone. Further investigation with these and other epidemiologic typing methods will be necessary to enhance the understanding of the epidemiology and pathogenesis of candidiasis in hospitalized patients.