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A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance

L Leach1, A Russell1, Y Zhu1

  • 1Mycology Laboratory, Wadsworth Center New York State Department of Health, University at Albany School of Public Health, Albany, New York, USA.

Journal of Clinical Microbiology
|August 9, 2019
PubMed

Insights

A new automated real-time PCR assay using the BD Max system efficiently detects Candida auris. This method enhances laboratory implementation for high-throughput screening of this multidrug-resistant yeast pathogen.

Area of Science:

  • Clinical microbiology
  • Molecular diagnostics
  • Infectious diseases

Background:

  • Candida auris is a multidrug-resistant yeast causing global outbreaks.
  • A previously developed real-time PCR assay aided C. auris outbreak investigations.
  • Need for an automated, easily implementable assay for C. auris detection.

Purpose of the Study:

  • To develop and optimize an automated sample-to-result real-time PCR assay for C. auris detection using the BD Max open system.
  • To improve laboratory workflow and high-throughput screening capabilities for C. auris surveillance.

Main Methods:

  • Optimized DNA extraction parameters (temperature and incubation time).
  • Validated sensitivity and specificity using patient samples and various yeast strains.
  • Assessed assay precision through independent extractions and replicate testing.

Main Results:

  • Established optimal BD Max assay conditions: DNA extraction at 75°C for 20 min with PerfeCTa multiplex qPCR ToughMix.
  • Achieved a limit of detection of one C. auris CFU/PCR.
  • Demonstrated 96% clinical sensitivity and 94% accuracy on 110 patient samples, detecting all four C. auris clades without cross-reactivity.

Conclusions:

  • The automated BD Max real-time PCR assay provides a precise, sensitive, and specific method for C. auris detection.
  • This high-throughput assay can screen 180 surveillance samples within a 12-hour workday.
  • Facilitates broader laboratory implementation for effective C. auris outbreak surveillance and control.
Keywords:
BD MaxCandida aurisautomatedhigh-throughputreal-time PCRsample-to-answersurveillance

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