Publication details

Rapid Identification of Medically Important Candida Isolates Using High Resolution Melting Analysis

Authors

NĚMCOVÁ Eva ČERNOCHOVÁ Michaela RŮŽIČKA Filip MALIŠOVÁ Barbora FREIBERGER Tomáš NĚMEC Petr

Year of publication 2015
Type Article in Periodical
Magazine / Source Plos one
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://www.plosone.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pone.0116940&representation=PDF
Doi http://dx.doi.org/10.1371/journal.pone.0116940
Field Epidemiology, infectious diseases and clinical immunology
Keywords REAL-TIME PCR; BLOOD CULTURE BOTTLES; POLYMERASE-CHAIN-REACTION; SPECIES IDENTIFICATION; ANTIFUNGAL SUSCEPTIBILITY; MOLECULAR-IDENTIFICATION; INVASIVE CANDIDIASIS; FUNGAL PATHOGENS; CURVE ANALYSIS; ACCURATE IDENTIFICATION
Attached files
Description 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.

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