Development and Validation of a Real-Time PCR Assay for Rapid Detection of Candida auris from Surveillance Samples

J Clin Microbiol. 2018 Jan 24;56(2):e01223-17. doi: 10.1128/JCM.01223-17. Print 2018 Feb.

Abstract

Candida auris is an emerging multidrug-resistant yeast causing invasive health care-associated infection with high mortality worldwide. Rapid identification of C. auris is of primary importance for the implementation of public health measures to control the spread of infection. To achieve these goals, we developed and validated a TaqMan-based real-time PCR assay targeting the internal transcribed spacer 2 (ITS2) region of the ribosomal gene. The assay was highly specific, reproducible, and sensitive, with the detection limit of 1 C. auris CFU/PCR. The performance of the C. auris real-time PCR assay was evaluated by using 623 surveillance samples, including 365 patient swabs and 258 environmental sponges. Real-time PCR yielded positive results from 49 swab and 58 sponge samples, with 89% and 100% clinical sensitivity with regard to their respective culture-positive results. The real-time PCR also detected C. auris DNA from 1% and 12% of swab and sponge samples with culture-negative results, indicating the presence of dead or culture-impaired C. auris The real-time PCR yielded results within 4 h of sample processing, compared to 4 to 14 days for culture, reducing turnaround time significantly. The new real-time PCR assay allows for accurate and rapid screening of C. auris and can increase effective control and prevention of this emerging multidrug-resistant fungal pathogen in health care facilities.

Keywords: Candida auris; TaqMan chemistry; assay validation; real-time PCR assay; surveillance samples.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Candida / genetics
  • Candida / isolation & purification*
  • Candidiasis / diagnosis*
  • Candidiasis / microbiology
  • DNA, Fungal / genetics
  • DNA, Ribosomal Spacer / genetics
  • Epidemiological Monitoring*
  • Female
  • Humans
  • Limit of Detection
  • Molecular Diagnostic Techniques / methods*
  • Real-Time Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Time Factors

Substances

  • DNA, Fungal
  • DNA, Ribosomal Spacer