Candida auris PCR for high-throughput infection control screening

Abstract

Candida auris has significant implications for infection control due to its multidrug resistance and spread in healthcare settings. Current culture-based screening methods are laborious and risks muco-cutaneous colonisation of laboratory staff. We describe the adaptation of a published real-time PCR for the identification of C. auris in skin swabs for high throughput infection control screening. Two published primer and probe sets were analysed utilising serial 10-fold dilutions of fifteen C. auris strains to assess the PCR limit of detection. One primer and probe set was compatible with our laboratory workflow and was selected for further development yielding a limit of detection of 1 colony forming unit (CFU) per reaction. Non-C. auris isolates, as well as routine skin swabs (n = 100) were tested by culture and PCR to assess specificity, where no cross-reactivity was detected. Skin swabs from a proven C. auris case (n = 6) were all both culture positive and PCR positive, while surveillance swabs from close contacts (n = 46) were all both culture negative and PCR negative. Finally, use of a lysis buffer comprising 4 M guanidinium thiocyanate rendered swab-equivalent quantities of C. auris non-viable, providing assurance of the safety benefit of PCR over culture. The development of a PCR assay for high throughput infection control screening is a promising method for rapid detection of C. auris with utility in an outbreak setting.

Development of a Novel Method for the Clinical Visualization and Rapid Identification of Multidrug-Resistant Candida auris

“…In addition to these advantages, the reaction time is also considerably reduced. The RAA-LFS detection system takes only 30 min from sample preparation to obtaining the results, saving valuable time for the treatment of critically ill patients while providing accurate results ( 42 , 43 ).…”

Strategies to Prevent Transmission of Candida auris in Healthcare Settings

“…Recent developments in molecular diagnostic procedures now allow rapid identification of invasive C. auris infections within a few hours with high sensitivity and specificity [ 52 , 57 , 97 ]. Quantitative real-time PCR assays have been developed for cost-effective and rapid detection of live C. auris cells from the hospital environment and from skin and other patient swab samples for the detection of colonized patients [ 98 – 100 ]. A portable quantitative real-time PCR-based point-of-care test for C. auris detection in clinical samples, reporting results in ≤ 30 min, has also been developed recently [ 101 ].…”