Abstract: Objective To develop a microtitration plate enzyme immunoassay to differentiate Candida albicans from Candida dubliniensis isolates using two specific probes simultaneously.Methods The fun-gus-specific universal primers derived from the internal transcribed spacer region of fungal rDNA were labeled with biotin,while the C.albicans or C.dubliniensis specific capture probes were coated on the microplates.Genomic DNA purified from the two species was amplified by PCR.The biotinylated products were captured by the probes coated on the microplates.The A₄₀₅ value was finally determined by the colorimetric assay.Results The two species of Candida could be detected specifically.Out of 108 clinical isolates originally identified as C.albicans on the basis of germtube formation,two isolates were positive for C.dubliniensis and negative for C.albicans.The other106isolates were positive for C.albicans and negative for C.dubliniensis.Conclusions Two-specific-probe hybridization method is rapid and reliable for differentiating C.albicans from C.dubliniensis.

Key words: Candida albicans, Candida dubliniensis, Nucleic acid hybridization, DNA probe