Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2025 May;68(5):e70065.
doi: 10.1111/myc.70065.

Performance Evaluation of Five Real-Time PCR Assays for the Detection of Candida auris DNA

Jochem B Buil  1   2 , Bart van den Bosch  1   2 , Suzan J van der Maas  1 , Eelco F J Meijer  2   3 , Theun de Groot  2   3 , Joseph Meletiadis  4   5   6 , Paul E Verweij  1   2   7 , Willem J G Melchers  1   2 , Suzan D Pas  1   2
Affiliations
Comparative Study

Performance Evaluation of Five Real-Time PCR Assays for the Detection of Candida auris DNA

Jochem B Buil et al. Mycoses. 2025 May.

Abstract

Objectives: This study aimed to systematically evaluate and compare the performance of two laboratory-developed assays (LDAs) and three commercially available real-time PCR assays for the detection of Candida auris. The analytical sensitivity, specificity and limit of detection (LOD) of each assay were assessed, alongside their clinical sensitivity in identifying C. auris colonisation.

Methods: Ten C. auris strains representing five clades, as well as genetically related yeasts, common yeast species, and dermatophytes, were used to assess assay sensitivity and cross reactivity. Clinical and environmental samples were collected from patients during an outbreak and tested with three commercial PCR assays (AurisID, Fungiplex, FungiXpert) and two LDAs (CDC LDA, EMC LDA). LOD was determined using Probit analysis. Diagnostic sensitivity was evaluated by comparing the detection rate of each individual assay to the total detection rate of all assays combined.

Results: The EMC LDA exhibited the highest analytical sensitivity, with a LOD of 8 conidia/reaction, followed by CDC LDA (16 conidia/reaction), AurisID and FungiXpert (19 conidia/reaction), and Fungiplex (596 conidia/reaction). Specificity testing revealed cross-reactivity in the CDC LDA and AurisID assays with C. pseudohaemulonii at high conidia levels, while no cross-reactivity was observed in the other assays. EMC LDA showed the highest clinical sensitivity (100%), whereas Fungiplex had the lowest positivity rate (71%). No false positives were observed in negative control swabs for any assay.

Conclusions: Real-time PCR is a crucial tool for the rapid and sensitive detection of C. auris , especially in clinical settings where timely identification is essential for effective patient management and infection control. Numerous PCR assays are available for this purpose; however, our study demonstrates that the sensitivity of these assays can vary significantly. The observed differences underscore the importance of establishing international reference standards and proficiency panels to enhance the accuracy and comparability of assay performance across different studies and laboratories.

Keywords: Candida; Candida spp; PCR.

PubMed Disclaimer

Conflict of interest statement

J.B.B. reports grants from Gilead Sciences, BioMérieux, lectures for Gilead Sciences and Pfizer, and advisory board for Gilead Sciences; all grants are paid to the institute. E.F.J.M. discloses speaker engagements for Gilead Sciences, advisory board fees from Pfizer, and research funding from MundiPharma and Scynexis. S.J.M. reports grants from Gilead Sciences and Mundipharma. P.E.V. reports consulting fees, advisory board and speaker fees from Gilead Sciences and consulting fees from Mundipharma, all paid to the institute. All authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Limit of detection analysis of C. auris PCR. Limit of detection is displayed with a 95% confidence interval. Due to low positivity, no 95% confidence interval could be calculated for the Fungiplex assay.
FIGURE 2
FIGURE 2
PCR positivity of patient and environmental samples and negative controls compared to any positive test and culture.
FIGURE 3
FIGURE 3
Comparative Cp‐values of five real‐time PCR assays for C. auris in culture‐positive and culture‐negative patients (A) and environmental (B) samples. −, C. auris culture negative samples; +, C. auris culture positive samples; LDA, Laboratory‐developed assay. Dots indicate individual Cp‐values, the horizontal line indicates the mean Cp‐value of a positive PCR. A Cp of 35 or above has higher uncertainty for the AurisID assay.
See this image and copyright information in PMC

References

    1. Chowdhary A., Sharma C., Duggal S., et al., “New Clonal Strain of Candida auris, Delhi, India,” Emerging Infectious Diseases 19, no. 10 (2013): 1670–1673, 10.3201/eid1910.130393. - DOI - PMC - PubMed
    1. Magobo R. E., Corcoran C., Seetharam S., and Govender N. P., “ Candida auris‐Associated Candidemia, South Africa,” Emerging Infectious Diseases 20, no. 7 (2014): 1250–1251, 10.3201/eid2007.131765. - DOI - PMC - PubMed
    1. Adams E., Quinn M., Tsay S., et al., “ Candida auris in Healthcare Facilities, New York, USA, 2013‐2017,” Emerging Infectious Diseases 24, no. 10 (2018): 1816–1824. - PMC - PubMed
    1. Satoh K., Makimura K., Hasumi Y., Nishiyama Y., Uchida K., and Yamaguchi H., “ Candida auris sp. Nov., a Novel Ascomycetous Yeast Isolated From the External Ear Canal of an Inpatient in a Japanese Hospital,” Microbiology and Immunology 53, no. 1 (2009): 41–44. - PubMed
    1. Chowdhary A., Sharma C., and Meis J. F., “ Candida auris: A Rapidly Emerging Cause of Hospital‐Acquired Multidrug‐Resistant Fungal Infections Globally,” PLoS Pathogens 13, no. 5 (2017): e1006290. - PMC - PubMed

MeSH terms

LinkOut - more resources