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

• Authors: Milena Kordalewska , Anna Brillowska-Dąbrowska , Tomasz Jagielski , Bożena Dworecka-Kaszak
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

Alternaria alternata; detection; identification; PCR; real-time PCR

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2015
• Volume: 62
• Issue: 4
• Pages: 707-712

Dates

published

2015

received

2015-07-30

revised

2015-10-02

accepted

2015-10-07

(unknown)

2015-10-27

Contributors

author

Milena Kordalewska

• Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, Gdańsk, Poland

author

Anna Brillowska-Dąbrowska

• Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, Gdańsk, Poland

author

Tomasz Jagielski

• Department of Applied Microbiology, Institute of Microbiology, Faculty of Biology, University of Warsaw, Warsaw, Poland

author

Bożena Dworecka-Kaszak

• Department of Applied Microbiology, Institute of Microbiology, Faculty of Biology, University of Warsaw, Warsaw, Poland

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Identifiers

DOI

10.18388/abp.2015_1112