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. 2018 Nov 27;56(12):e01337-18.
doi: 10.1128/JCM.01337-18. Print 2018 Dec.

Direct Detection of Emergent Fungal Pathogen Candida auris in Clinical Skin Swabs by SYBR Green-Based Quantitative PCR Assay

D Joseph Sexton  1 , Milena Kordalewska  2 , Meghan L Bentz  3 , Rory M Welsh  3 , David S Perlin  2 , Anastasia P Litvintseva  1
Affiliations

Direct Detection of Emergent Fungal Pathogen Candida auris in Clinical Skin Swabs by SYBR Green-Based Quantitative PCR Assay

D Joseph Sexton et al. J Clin Microbiol. .

Abstract

The recent emergence of the multidrug-resistant and pathogenic yeast Candida auris continues to cause public health concern worldwide. C. auris is alarming because it causes health care-associated outbreaks and can establish invasive infections with high mortality rates. Transmission between patients is facilitated by the ability of C. auris to persistently colonize multiple body sites, including the skin, and survive for weeks on surfaces in health care settings. Rapid identification of colonized patients is needed to implement timely infection control measures. Currently, CDC laboratories use an enrichment culture-based approach that can take up to 2 weeks to identify C. auris from composite swabs from the bilateral axillae and groin. A rapid SYBR green quantitative PCR (qPCR) assay that can identify C. auris in a single day was recently described. In this study, we developed the SYBR green qPCR assay further by incorporating a DNA extraction procedure for skin swabs and by including an internal amplification control based on the distinguishable melt curve of a lambda DNA amplicon. The assay was conducted using 103 clinical axilla/groin skin swab samples. Using the enrichment culture-based approach as a gold standard, we determined that the SYBR green C. auris qPCR has a sensitivity of 0.93 and specificity of 0.96. Overall, we found that the SYBR green C. auris qPCR assay can be successfully applied for rapid and accurate detection of C. auris in patient skin swabs, thereby increasing diagnostic options for this emerging pathogen.

Keywords: Candida auris; SYBR; diagnostic.

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Figures

FIG 1
FIG 1
Melt curve analysis of two reactions that both contain 0.01 ng of C. auris gDNA. The original qPCR master mix includes only C. auris primers and is shown by the dotted line. The new master mix also includes lambda DNA and primers serving as an internal amplification control (IAC), shown with the solid line. The presence of C. auris is indicated by a melt peak at 83.8°C, and the presence of lambda DNA is indicated by a melt peak at 88.4°C.
FIG 2
FIG 2
Melting curve analysis of 103 clinical swabs samples (left) and the ROC curve analysis (right) that was performed to establish the optimum threshold at 245 −d(RFU)/dT. Left, the presence of C. auris is indicated by a melt peak at 83.8°C, and the presence of lambda DNA is indicated by a melt peak at 88.4°C.
See this image and copyright information in PMC

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