Rapid Identification of Seven Waterborne Exophiala Species by RCA DNA Padlock Probes

M. J. Najafzadeh, V. A. Vicente, Peiying Feng, A. Naseri, Jiufeng Sun, A. Rezaei-Matehkolaei, G. S. de Hoog

Research output: Chapter in book/volumeChapterScientificpeer-review

Abstract

The black yeast genus Exophiala includes numerous potential opportunistic species that potentially cause systematic and disseminated infections in immunocompetent individuals. Species causing systemic disease have ability to grow at 37–40 °C, while others consistently lack thermotolerance and are involved in diseases of cold-blooded, waterborne vertebrates and occasionally invertebrates. We explain a fast and sensitive assay for recognition and identification of waterborne Exophiala species without sequencing. The ITS rDNA region of seven Exophiala species (E. equina, E. salmonis, E. opportunistica, E. pisciphila, E. aquamarina, E. angulospora and E. castellanii) along with the close relative Veronaea botryosa was sequenced and aligned for the design of specific padlock probes for the detection of characteristic single-nucleotide polymorphisms. The assay demonstrated to successfully amplify DNA of target fungi, allowing detection at the species level. Amplification products were visualized on 1% agarose gels to confirm specificity of probe–template binding. Amounts of reagents were reduced to prevent the generation of false positive results. The simplicity, tenderness, robustness and low expenses provide padlock probe assay (RCA) a definite place as a very practical method among isothermal approaches for DNA diagnostics. © 2018 The Author(s)
Original languageEnglish
Title of host publicationMycopathologia
PublisherSpringer Netherlands
Pages669-677
Number of pages9
ISBN (Print)0301486X (ISSN)
DOIs
Publication statusPublished - 01 Aug 2018

Publication series

NameMycopathologia
Volume183

Keywords

  • Black yeasts
  • Identification
  • Rolling circle amplification
  • Waterborne Exophiala

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    Najafzadeh, M. J., Vicente, V. A., Feng, P., Naseri, A., Sun, J., Rezaei-Matehkolaei, A., & de Hoog, G. S. (2018). Rapid Identification of Seven Waterborne Exophiala Species by RCA DNA Padlock Probes. In Mycopathologia (pp. 669-677). (Mycopathologia; Vol. 183). Springer Netherlands. https://doi.org/10.1007/s11046-018-0256-7