An immunogenic aminopeptidase of Candida albicans was purified by high-performance liquid chromatography. It was then used for the development of an enzyme-linked immunosorbent assay to detect antibodies directed against this antigen in sera from patients with candidiasis. This enzyme specifically cleaves the L-Arg-7-amino-4-methyl-coumarin substrate at pH 7.4 and was detected in the crude extract of different C. albicans isolates. Sera used for this study were obtained from healthy blood donors or from patients with one of the following: systemic candidiasis, aspergillosis, cryptococcosis, toxoplasmosis, or malaria. The statistical analysis demonstrates significant differences between absorbency values obtained with sera from patients with candidiasis and with sera from the other groups (P = 0.000001). Diagnostic parameters show high diagnostic specificity of 97% and a sensitivity of 83% at a cutoff value of 0.425 and suggest the usefulness of this aminopeptidase for the diagnosis of systemic candidiasis.