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A high-throughput and rapid method for accurate identification of emerging multidrug-resistant Candida auris

Ausaf Ahmad1, Jonathan E Spencer1, Shawn R Lockhart2

  • 1Enhanced Capacity Surge Laboratory, Reagent and Diagnostic Services Branch, Division of Scientific Resources, National Center for Emerging and Zoonotic Infectious Diseases (NCEZID) Centers for Disease Control & Prevention (CDC), Atlanta, Georgia.

Mycoses
|February 26, 2019
PubMed

Insights

A new rapid DNA extraction and real-time PCR method accurately identifies the multidrug-resistant yeast Candida auris (C. auris). This high-throughput assay significantly reduces turnaround time for clinical and public health applications.

Area of Science:

  • Medical Mycology
  • Infectious Diseases
  • Molecular Diagnostics

Background:

  • Candida auris is an emerging multidrug-resistant yeast causing invasive infections in healthcare settings.
  • Increasing C. auris cases in the US necessitate rapid diagnostic tools for patient care and infection control.
  • Accurate and timely identification of C. auris is crucial for public health interventions.

Purpose of the Study:

  • To develop and validate a rapid DNA extraction method using the Roche MagNA Pure 96 instrument.
  • To create and validate a TaqMan real-time PCR assay for high-throughput C. auris identification.
  • To assess the diagnostic accuracy and efficiency of the developed method.

Main Methods:

  • Evaluated 247 patient dermal swab samples previously analyzed by culture/MALDI-TOF.
  • Utilized Roche MagNA Pure 96 for DNA extraction and TaqMan real-time PCR for C. auris detection.
  • Performed receiver operating characteristic (ROC) curve analysis to determine optimal cut-off values.

Main Results:

  • The assay demonstrated high diagnostic sensitivity (93.6%) and specificity (97.2%).
  • Achieved a low limit of detection (1 C. auris CFU/10 µL) and high reproducibility.
  • The method offers a high throughput of approximately 200 samples per day with a significantly reduced turnaround time.

Conclusions:

  • A rapid, high-throughput DNA extraction and real-time PCR method for C. auris identification was successfully validated.
  • The developed assay provides accurate and reproducible results, outperforming traditional methods in terms of speed.
  • This method is suitable for clinical laboratories and public health surveillance of C. auris infections.
Keywords:
Candida aurisTaqMan real-time PCRhigh-throughput rapid DNA extraction

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