Internal validation of GPS^™MONODOSE CanAur dtec‐qPCR kit following the UNE/EN ISO/IEC 17025:2005 for detection of the emerging yeast Candida auris

Abstract

Candida auris is an emerging multidrug resistant pathogenic fungus that causes candidaemia with high mortality rates and exhibits the ability to persist within the hospital environment. Candida auris is phylogenetically closely related to Candida haemulonii, C. lusitaniae, C. pseudohaemulonii and C. duobushaemulonii and is frequently misidentified by commercial identification methods. In the present study, the GPS MONODOSE dtec-qPCR kit (dried single-dose PCR tubes) for the detection of C. auris was validated following the guidelines of the UNE/EN ISO/IEC 17025:2005, the French Standard NF T90-471:2010 and using fast-cycling protocols. Validation terms included in vitro specificity (inclusivity/exclusivity), quantitative phase analysis (10-10 standard DNA copies), reliability (repeatability/reproducibility) and sensitivity (detection/quantification limits). GPS dtec-qPCR kits passed validation with strict acceptance criteria (n ≥ 10 repetitions). In silico specificity was proven by the designer (GPS ). Experimental inclusiveness was achieved in two independent laboratories by testing strain JCM15448 and 117 clinical C. auris isolates from Asia, the Middle-East Africa, Latin America and Europe. Exclusiveness was evaluated with 25 strains of closely related Candida spp. Use of the MONODOSE format provided considerable advantages allowing the detection of C. auris to be accurately achieved in less than an hour. The GPS MONODOSE dtec-qPCR kit is ready to undergo clinical evaluation.

Candida auris: A Review of Recommendations for Detection and Control in Healthcare Settings

“…At the time of writing, clinical evaluations of three culture-independent tests using culture-based gold standards have been published. These include a Taqman quantitative PCR (qPCR), a SYBR green qPCR, and a T2 Magnetic Resonance assay; all performed well with clinical sensitivities and specificities close to or exceeding 90% [19][20][21][22][23]. In the United States, the Taqman-based qPCR is currently the mostly widely used culture-independent test and is employed for C. auris surveillance at CDC and the Wadsworth Center in New York, as well as an increasing number of Antibiotic Resistance Laboratory Network (AR Lab Network) laboratories.…”

Antifungal Resistance: Specific Focus on Multidrug Resistance in Candida auris and Secondary Azole Resistance in Aspergillus fumigatus

“…It is therefore of utmost importance to quickly identify contaminated surfaces and screen specimens of patients. Real-time detection and identification of C. auris is the target of several molecular kits [51,52,53,54,55,56,57,58,59]. The survival of C. auris for weeks, even months, within the hospital confirms the importance of infection prevention programs [60,61,62].…”

Candida auris—the growing menace to global health

“…Recently, GPS MONODOSE dtec‐qPCR kit (Alicante, Spain) for the detection of C. auris have been introduced with many advantages compared to other PCR methods. This kit is user friendly and detects the target in less than an hour …”