Rapid identification of Candida glabrata cryptic species using real-time PCR combined with denaturing highperformance liquid chromatography

Ref ID: 17731

Author:

O. Telleria, G. Ezpeleta*, S. Hernaez, R. Cisterna

Author address:

Bilbao, ES)

Full conference title:

22nd European Congress of Clinical Microbiology and Infectious Diseases

Abstract:

Introduction: During last years, the increment in incidence, and
associated high morbi-mortality, has converted invasive fungal
infections in one of the most important public health associated
problems. Besides, non-albicans Candida species have emerged as
etiological agents of invasive candidiasis. Since the description of two
new cryptic species (C. bracarensis and C. nivariensis)
phylogenetically related to C. glabrata with different phenotype and
antifungal susceptibility profile, it seems to be necessary to develop a
rapid and accurate identification technique in order to distinguish
between these three microorganisms.
Objective: We studied the performance of real-time PCR combined
with Denaturing High Liquid Chromatography (DHPLC) as an
alternative, fast and novel method to perform such identification
accurately.
Methodology and results: Fungal DNA from pure cultures of C.
glabrata (04.229), C. bracarensis (NCYC-3133) and C. nivariensis
(04.228) reference strains was extracted using a MagNAPure Compact
system. A small amplicon of the ITS2 region was amplified using the
ITS86-F and ITS4-R primers previously described by Grutzner et al.
employing a real-time PCR scheme with SYBR Green I in a
LightCycler 2.0. The PCR products obtained were purified using

Abstract Number: NULL

Conference Year: 2012

Link to conference website: NULL

New link: NULL

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