Mycoses. 2018;61:877–884. wileyonlinelibrary.com/journal/myc
|
877 © 2018 Blackwell Verlag GmbH
1 | INTRODUCTION
The multidrug-resistant ascomycete yeast pathogen Candida auris
was first described from Japan, followed by South Korea in the
same year.
1,2
This new species was closely related to Candida lu-
sitaniae, C. pseudohaemulonii, C. duobushaemulonii and C. haem-
ulonii, known as the C. haemulonii species complex.
3,4
Currently,
C. auris has been reported in >25 countries on five continents:
North America, South America, Europe, Africa, and Asia.
5,6
Unlike
other yeasts, they are transmitted within and between hospi-
tals, posing a serous public health threat.
7
Early detection of
C. auris infections may be beneficial for early initiation of appro-
priate antifungal therapy and prevention of cross infections.
8-12
Nevertheless, the inability of most available commercial identifi-
cation methods to quickly diagnose C. auris remains a drawback
to early therapy.
13
There are currently no therapeutic guidelines,
Received: 18 May 2018
|
Revised: 23 July 2018
|
Accepted: 23 July 2018
DOI: 10.1111/myc.12834
ORIGINAL ARTICLE
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
Antonio Martínez-Murcia
1,2
| Aaron Navarro
2
| Gema Bru
2
| Anuradha Chowdhary
3
|
Ferry Hagen
4,5
| Jacques F. Meis
4,6
1
Department of Microbiology, University
Miguel Hernández, Orihuela, Alicante, Spain
2
Genetic PCR Solutions™, Elche, Alicante,
Spain
3
Department of Medical Mycology,
Vallabhbhai Patel Chest Institute, University
of Delhi, Delhi, India
4
Department of Medical Microbiology and
Infectious Diseases, ECMM Excellence
Center, Canisius-Wilhelmina Hospital (CWZ),
Nijmegen, The Netherlands
5
Department of Medical Mycology,
Westerdijk Fungal Biodiversity Institute,
Utrecht, The Netherlands
6
Centre of Expertise in Mycology
Radboudumc/CWZ, Nijmegen, The
Netherlands
Correspondence: Antonio Martínez-Murcia,
Genetic PCR Solutions™, CEEI – Ronda Vall
d’Uxó 125, 03206-Elche (Alicante) Spain
(ammurcia@geneticpcr.com).
Funding information
European Union Seventh Framework
Programme [FP7/2007-2011], Grant/
Award Number: 311846; IVACE, Generalitat
Valenciana, Grant/Award Number:
IMIDTA/2018/16
Summary
Candida auris is an emerging multidrug resistant pathogenic fungus that causes can-
didaemia with high mortality rates and exhibits the ability to persist within the hos-
pital environment. Candida auris is phylogenetically closely related to Candida
haemulonii, C. lusitaniae, C. pseudohaemulonii and C. duobushaemulonii and is fre-
quently 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
6
standard DNA copies), reliability (repeatability/reproducibility) and sensitiv-
ity (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 labo-
ratories by testing strain JCM15448
T
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 con-
siderable 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.
KEYWORDS
Candida auris, detection, qPCR, UNE/EN ISO/IEC 17025:2005, validation