The Aspergillus galactomannan enzyme-linked immunosorbant assay (EIA) has been demonstrated to facilitate rapid and sensitive detection of invasive aspergillosis. However, test specificity has not been fully evaluated in non-Aspergillus fungal species. Of 53 fungal isolates, cross-reactivity was observed with 5 non-Aspergillus spp.: Blastomyces dermatitidis, Nigrospora oryzae, Paecilomyces lilacinus, Penicillium chrysogenum, and Trichothecium roseum.