PCR and real-time PCR assays to detect fungi of Alternaria alternata species

Milena Kordalewska; Anna Brillowska-Dąbrowska; Tomasz Jagielski; Bożena Dworecka-Kaszak

doi:10.18388/abp.2015_1112

PCR and real-time PCR assays to detect fungi of Alternaria alternata species

Abstract

Fungi of the Alternaria genus are mostly associated with allergic diseases. However, with a growing number of immunocompromised patients, these fungi, with A. alternata being the most prevalent one, are increasingly recognized as etiological agents of infections (phaeohyphomycoses) in humans. Nowadays, identification of Alternaria spp. requires their pure culture and is solely based on morphological criteria. Clinically, Alternaria infections may be indistinguishable from other fungal diseases. Therefore, a diagnostic result is often delayed or even not achieved at all. In this paper we present easy to perform and interpret PCR and real-time PCR assays enabling detection of A. alternata species. On the basis of alignment of β-tubulin gene sequences, A. alternata-specific primers were designed. DNA from fungal isolates, extracted in a two-step procedure, were used in PCR and real-time PCR assays followed by electrophoresis or melting temperature analysis, respectively. The assays specificity was confirmed, since positive results were obtained for all A. alternata isolates, and no positive results were obtained neither for other molds, dermatophytes, yeast-like fungi, nor human DNA. The assays developed here enable fast and unambiguous identification of A. alternata pathogens.

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Authors (4)

Milena Kordalewska
Anna Brillowska-Dąbrowska dr hab. inż.
Tomasz Jagielski dr
- UW Zakład Mikrobiologii Stosowanej
Bożena Dworecka-Kaszak
- SGGW Katedra Nauk Przedklinicznych

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DOI:: Digital Object Identifier (open in new tab) 10.18388/abp.2015_1112
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Category:: Articles
Type:: artykuł w czasopiśmie wyróżnionym w JCR
Published in:: Acta Biochimica Polonica no. 62, pages 707 - 712,
ISSN: 0001-527X
Language:: English
Publication year:: 2015
Bibliographic description:: Kordalewska M., Brillowska-Dąbrowska A., Jagielski T., Dworecka-Kaszak B.: PCR and real-time PCR assays to detect fungi of Alternaria alternata species// Acta Biochimica Polonica. -Vol. 62, iss. 4 (2015), s.707-712
DOI:: Digital Object Identifier (open in new tab) 10.18388/abp.2015_1112
Verified by:: Gdańsk University of Technology