Candida dubliniensis is a pathogenic yeast of the genus Candida closely related to Candida albicans. The phenotypic similarity of these two species often leads to misidentification of C. dubliniensis isolates in clinical samples. DNA-based methods continue to be the most effective means of discriminating accurately between the two species. Here, we report on the identification of nine unusual Candida isolates that showed ambiguous identification patterns on the basis of their phenotypic and immunological traits. The isolates were categorized into two groups. Group I isolates were unable to produce germ tubes and chlamydospores, and to agglutinate commercial latex particles coated with a mAb highly specific for C. dubliniensis. Group II isolates grew as pink and white colonies on CHROMagar Candida and ChromID Candida, respectively. Carbohydrate assimilation profiles obtained with API/ID32C together with PCR amplification with specific primers and DNA sequencing allowed reliable identification of the nine unusual clinical isolates as C. dubliniensis.