Development and Analysis of qPCR for the Identification of Arthroconidial Yeasts of the Genus Magnusiomyces

Engin Kaplan, Deniz Aktaş, Aylin Döğen, Süleyha Hilmioğlu-Polat, Ramazan Gümral, Ferry Hagen, Macit Ilkit, G Sybren de Hoog

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)41-51
Aantal pagina's11
TijdschriftMycopathologia
Volume186
Nummer van het tijdschrift1
DOI's
StatusGepubliceerd - mrt 2021

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