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

Rev Iberoam Micol. Apr-Jun 2018;35(2):110-112. doi: 10.1016/j.riam.2018.01.003. Epub 2018 Apr 21.

Abstract

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.

Keywords: Candida auris; Candida haemulonii; Identificación molecular; Molecular identification.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Base Sequence
  • Candida / classification*
  • Candida / genetics
  • Candidiasis / microbiology
  • Communicable Diseases, Emerging / microbiology
  • DNA Primers
  • DNA, Fungal / genetics
  • DNA, Ribosomal Spacer / genetics
  • Humans
  • Multiplex Polymerase Chain Reaction / instrumentation
  • Multiplex Polymerase Chain Reaction / methods*
  • Mycological Typing Techniques / methods*
  • Sequence Analysis, DNA
  • Species Specificity
  • Yeasts / genetics

Substances

  • DNA Primers
  • DNA, Fungal
  • DNA, Ribosomal Spacer