Validation of the PCR–dHPLC method for rapid identification of Candida glabrata phylogenetically related species in different biological matrices

Since two new species phylogenetically related to Candida glabrata with slightly different phenotypes and antifungal susceptibility profiles have been described, it seems to be necessary from clinical point of view, to develop a rapid and accurate identification system in order to distinguish between these three fungal species. We studied the performance of denaturing high performance liquid chromatography (dHPLC) as a faster (less than 7 min) and alternative novel technique for simultaneous analysis of Candida species in different biological matrices. The analyses show the good low limit of detection (LLOD) in all biological matrices studied (5.16–9.56 ng μL−1, 4.14–4.70 ng μL−1 and 3.99–4.66 ng μL−1 for Candida bracarensis, Candida nivariensis and C. glabrata, respectively). 180 Candida isolates were analyzed in order to demonstrate the method suitability for screening analysis to identify C. glabrata and its cryptic species (C. bracarensis and C. nivariensis) in clinical routine.

► We tried to differentiate two new species (C. bracarensis and C. nivariensis) close related to C. glabrata with different phenotype and antifungal susceptibility profile.
► We studied the performance of denaturing high performance liquid chromatography (dHPLC) as a fast and alternative novel technique for simultaneous identification of these three Candida species in different biological matrices.
► dHPLC is a suitable method for screening analysis to identify C. glabrata and its cryptic species.