Single-tube classical PCR for Candida auris and Candida haemulonii identification

Background: Candida auris and Candida haemulonii are emerging and multiresistant pathogens. C auris has produced hospital outbreaks and is misidentified by phenotypic-based methods. The only reliable identification methods are DNA sequencing and MALDI-TOF. Aims: To develop a classical-PCR method capable of rapidly and accurately identify C. auris and C haemulonii. Methods: A multiplex PCR was carried out in one tube that included an internal control and oligonucleotides that specifically hybridize to the ITS2 region of C auris and C. haemulonii. The usefulness of the new method was verified by testing a collection of 50 strains of 20 different species (previously identified by ITS sequencing). The selection of species was made in order to emulate the C auris panel used by the CDC to validate diagnostic tools. In addition, other yeast species not included in the aforementioned panel were incorporated based on reported identification errors. Results: The results obtained with the proposed protocol were in total agreement with those obtained by ITS sequencing. Conclusions: We present a PCR method able to unequivocally identify C. auris and differentiate it from C. haemulonii. It is inexpensive, fast and it could be a useful tool to reduce the chances of a C. auris outbreak. (C) 2018 Asociacion Espanola de Micologia. Published by Elsevier Espana, S.L.U. All rights reserved.