Multilocus enzyme electrophoresis was adapted for subtyping Cryptococcus neoformans. The two cryptococcal varieties were clearly distinguishable. Isolates of the C. neoformans var. neoformans were sorted according to serotype and were sorted into four to five subtypes within each serotype. Nearly no two isolates of the C. neoformans var. gattii displayed the same enzyme electrophoretic type. This method may be a useful adjunct to current methods for classification and epidemiologic studies of cryptococci.