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PLoS One. 2015 Feb 17;10(2):e0116940. doi: 10.1371/journal.pone.0116940. eCollection 2015.

Rapid identification of medically important Candida isolates using high resolution melting analysis.

Author information

1
Centre for Cardiovascular Surgery and Transplantation Brno, Brno, Czech Republic; International Clinical Research Centre, St. Anne's University Hospital Brno, Brno, Czech Republic.
2
Centre for Cardiovascular Surgery and Transplantation Brno, Brno, Czech Republic; Molecular Immunology and Microbiology, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
3
Department of Microbiology, Faculty of Medicine, Masaryk University and St. Anne's University Hospital Brno, Brno, Czech Republic.
4
Molecular Immunology and Microbiology, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.

Abstract

An increasing trend in non albicans infections and various susceptibility patterns to antifungal agents implies a requirement for the quick and reliable identification of a number of medically important Candida species. Real-time PCR followed by high resolution melting analysis (HRMA) was developed, tested on 25 reference Candida collection strains and validated on an additional 143 clinical isolates in this study. All reference strains and clinical isolates inconclusive when using phenotypic methods and/or HRMA were analysed using ITS2 sequencing. Considering reference and clinical strains together, 23 out of 27 Candida species could be clearly distinguished by HRMA, while the remaining 4 species were grouped in 2 pairs, when applying the mean Tm ± 3 SD values, the shape of the derivative melting curve (dMelt curve) and, in some cases, the normalized and temperature-shifted difference plot against C. krusei. HRMA as a simple, rapid and inexpensive tool was shown to be useful in identifying a wide spectrum of clinically important Candida species. It may complement the current clinical diagnostic approach based on commercially available biochemical kits.

PMID:
25689781
PMCID:
PMC4331273
DOI:
10.1371/journal.pone.0116940
[Indexed for MEDLINE]
Free PMC Article

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