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Development and Analysis of qPCR for the Identification of Arthroconidial Yeasts of the Genus Magnusiomyces |
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| Authors: | Kaplan Engin Akta? Deniz Dö?en Aylin Hilmio?lu-Polat Süleyha Gümral Ramazan Hagen Ferry Ilkit Macit de Hoog G Sybren |
| Institution: | 1.Department of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Zonguldak Bülent Ecevit, Zonguldak, Turkey ;2.Department of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Mersin, Mersin, Turkey ;3.Department of Microbiology, Faculty of Medicine, University of Ege, Izmir, Turkey ;4.Department of Microbiology, Gülhane Military Medical Academy, Ankara, Turkey ;5.Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands ;6.Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands ;7.Laboratory of Medical Mycology, Jining No. 1, People’s Hospital, Jining, Shandong, P.R. China ;8.Division of Mycology, Department of Microbiology, Faculty of Medicine, University of ?ukurova, Adana, Turkey ;9.Centre of Expertise in Mycology, Radboud University Medical Centre/Canisius Wilhelmina Hospital, Nijmegen, The Netherlands ; |
| Abstract: | The arthroconidial yeasts Magnusiomyces capitatus and M. clavatus are emerging opportunistic pulmonary pathogens. They are closely related and difficult to distinguish based on morphological and physiological traits. We applied an SYBR® green-based quantitative PCR (qPCR) assay to identify the species. We analyzed 30 reference strains originating from clinical and environmental sources by targeting the Rpb2 gene encoding the second largest subunit of RNA polymerase II. The qPCR assays were tested by direct identification of M. capitatus and M. clavatus in spiked sputum and household dishwasher swabs, respectively, as models for clinical and environmental samples. The assays were proved to be reliable for species-level identification of both species, with 100% sensitivity and 100% specificity, lowest inter-assay deviations (RSDr?≤?1.65%, R2 values >0.99), detection limit of 10 theoretical copy number of target DNA, and detection cell limit of ≥5000 yeast cells from spiked sputum samples. The developed qPCR assay is a practical molecular approach for the detection of M. capitatus and M. clavatus that can be used as a stand-alone assay or in conjunction with culture-dependent approaches. |
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