Identification of Candida and Aspergillus species using a new multiplex PCR-based methodology

Ref ID: 19389

Author:

C. O. Vaz,1 J. I. Carvalho-Pereira,1 R. Ara ujo,2 C. S. S. Pais1 and
P. F. M. Sampaio1

Author address:

1University of Minho, Braga, Portugal and 2IPATIMUP, Institute of
Molecular Pathology and Immunology, University of Porto,
Portugal

Full conference title:

6th Trends in Medical Mycology 2013

Date: 11 October 2014

Abstract:

Fungal pathogens are the major eukaryotic agents of serious infec-
tion in Europe, between them, Candida spp. and Aspergillus spp. repre-
sent the most common and clinically important etiological agents.
Due to their different susceptibility to antifungal drugs, the rapid and
correct identification of these infecting species is crucial. Several
methods have been developed for species identification, but they still
present several limitations, such as low specificity and sensibility. In
order to overcome these limitations, the analysis of microsatellite
sequences represents an excellent alternative for the differentiation
and characterization of clinically important species.
In this way, the main purpose of this study is the development of
a multiplex PCR strategy for the identification of the major Candida
spp. and Aspergillus spp. species. In order to accomplish this, markers
for C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, A. fu-
migatus, A. flavus, A. niger and A. terreus were selected for loci specific
amplification and were combined into a multiplex strategy. This new
methodology was tested in 120 strains from 15 different species for
specificity, sensibility and reproducibility.
All the selected loci were tested in single and multiplex conditions
and they have demonstrated 100% of specificity and sensibility. This
multiplex system developed showed to be a fast, accurate and repro-
ducible method, allowing the accurate identification of all 9 fungal
species. The methodology developed is easy to perform and can be
implemented at relatively low cost for routine identification in micro-
biology laboratories.

Abstract Number: NULL

Conference Year: 2013

Link to conference website: NULL

New link: NULL

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