Development and Analysis of qPCR for the Identification of Arthroconidial Yeasts of the Genus Magnusiomyces
Küçük Resim Yok
Tarih
2021
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Springer Science and Business Media B.V.
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
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. © 2021, The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature.
Açıklama
Anahtar Kelimeler
Arthroconidial yeasts, Molecular diagnostics, Quantitative real-time PCR
Kaynak
Mycopathologia
WoS Q Değeri
Scopus Q Değeri
N/A
Cilt
186
Sayı
1
