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Comparison of Two Commercially Available qPCR Kits for the Detection of Candida auris
Janko Sattler1,2, Janina Noster3, Anne Brunke1,2
1Institute for Medical Microbiology, Immunology and Hygiene, University Hospital of Cologne, University of Cologne, 50935 Cologne, Germany.
Abstract:
Candida auris is an emerging pathogen with resistance to many commonly used antifungal agents. Infections with C. auris require rapid and reliable detection methods to initiate successful medical treatment and contain hospital outbreaks. Conventional identification methods are prone to errors and can lead to misidentifications. PCR-based assays, in turn, can provide reliable results with low turnaround times. However, only limited data are available on the performance of commercially available assays for C. auris detection. In the present study, the two commercially available PCR assays AurisID (OLM, Newcastle Upon Tyne, UK) and Fungiplex Candida Auris RUO Real-Time PCR (Bruker, Bremen, Germany) were challenged with 29 C. auris isolates from all five clades and eight other Candida species as controls. AurisID reliably detected C. auris with a limit of detection (LoD) of 1 genome copies/reaction. However, false positive results were obtained with high DNA amounts of the closely related species C. haemulonii, C. duobushaemulonii and C. pseudohaemulonii. The Fungiplex Candida Auris RUO Real-Time PCR kit detected C. auris with an LoD of 9 copies/reaction. No false positive results were obtained with this assay. In addition, C. auris could also be detected in human blood samples spiked with pure fungal cultures by both kits. In summary, both kits could detect C. auris-DNA at low DNA concentrations but differed slightly in their limits of detection and specificity.
Insights
Two commercial PCR assays were evaluated for detecting the emerging fungal pathogen Candida auris. Both kits detected C. auris DNA, but differed in sensitivity and specificity, impacting clinical diagnosis and outbreak control.
Area of Science:
- Clinical Microbiology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Candida auris is a multidrug-resistant emerging pathogen causing severe infections.
- Rapid and accurate detection of C. auris is crucial for treatment and outbreak containment.
- Conventional identification methods are often slow and error-prone, necessitating molecular approaches.
Purpose of the Study:
- To evaluate the performance of two commercially available PCR assays for C. auris detection.
- To compare the limit of detection and specificity of the AurisID and Fungiplex Candida Auris RUO Real-Time PCR kits.
- To assess the utility of these assays in detecting C. auris in spiked human blood samples.
Main Methods:
- Two commercial PCR assays, AurisID and Fungiplex Candida Auris RUO Real-Time PCR, were tested.
- A panel of 29 C. auris isolates from five clades and eight other Candida species were used.
- Limit of detection and specificity were determined, and assays were challenged with spiked human blood samples.
Main Results:
- AurisID detected C. auris with a limit of detection of 1 genome copy/reaction but showed false positives with high DNA amounts of related species.
- Fungiplex Candida Auris RUO Real-Time PCR detected C. auris with a limit of detection of 9 copies/reaction and exhibited no false positives.
- Both assays successfully detected C. auris DNA in spiked human blood samples.
Conclusions:
- Both evaluated PCR kits can detect C. auris DNA at low concentrations.
- The Fungiplex assay demonstrated superior specificity compared to AurisID.
- These findings highlight the importance of assay selection for accurate C. auris diagnosis and surveillance.
