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

Najafzadeh, M. J. and Vicente, V. A. and Feng, P. and Naseri, A. and Sun, J. and Rezaei-Matehkolaei, A. and de Hoog, G. S. (2018) Rapid Identification of Seven Waterborne Exophiala Species by RCA DNA Padlock Probes. Mycopathologia, 183 (4). pp. 669-677.

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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).

Item Type: Article
Additional Information: Cited By :1 Export Date: 16 February 2020 CODEN: MYCPA Correspondence Address: de Hoog, G.S.; Microbiology, Parasitology and Pathology Post-Graduation Program, Department of Basic Pathology, Federal University of ParanáBrazil; email: s.hoog@westerdijkinstitute.nl
Uncontrolled Keywords: Black yeasts Identification Rolling circle amplification Waterborne Exophiala fungal DNA ribosomal spacer DNA chemistry classification DNA sequence Exophiala genetics isolation and purification microbiological examination microbiology molecular diagnosis nucleic acid amplification oligonucleotide probe procedures sensitivity and specificity DNA, Fungal DNA, Ribosomal Spacer Microbiological Techniques Molecular Diagnostic Techniques Nucleic Acid Amplification Techniques Oligonucleotide Probes Sequence Analysis, DNA Water Microbiology
Subjects: QW Microbiology and Immunology
QX Parasitology
Divisions: Mashhad University of Medical Sciences
Depositing User: lib2 lib2 lib2
Date Deposited: 07 May 2020 13:37
Last Modified: 07 May 2020 13:37
URI: http://eprints.mums.ac.ir/id/eprint/17443

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