Multiplex polymerase chain reaction assay for simultaneous detection of Candida albicans and Candida dubliniensis

A multiplex polymerase chain reaction (PCR) assay was developed for the identification of two Candida species-albicans and dubliniensis. Three sets of primers were selected from different genomic sequences to specifically amplify a 516 bp fragment within the top2 gene, specific for several species of the genus Candida (CCL primers); a 239 bp fragment within the alphaINT gene, specific for Candida albicans (CAL primers); and a 175 bp fragment within the ALSDI gene, specific for Candida dubliniensis (CDL primers). Using the primers in conjunction (multiplex PCR), we were able to detect both C. albicans and C. dubliniensis and to differentiate between them. The detection limit of the PCR assay was approximately 10 cells per milliliter of saline. Thus, this multiplex PCR assay can be applied for differentiation of C. albicans and C. dubliniensis from clinical specimens.