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Journal of Clinical Microbiology, April 2009, p. 1063-1073, Vol. 47, No. 4
0095-1137/09/$08.00+0 doi:10.1128/JCM.01558-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Species-Specific Identification of a Wide Range of Clinically Relevant Fungal Pathogens by Use of Luminex xMAP Technology
,
C. Landlinger,1,2
S. Preuner,1
B. Willinger,3
B. Haberpursch,4
Z. Racil,5
J. Mayer,5 and
T. Lion1,2*
Division of Molecular Microbiology and Development of Genetic Diagnostics, Children's Cancer Research Institute,1 LabDia Labordiagnostik GmbH, Vienna, Austria,2 Division of Clinical Microbiology, Institute of Hygiene and Medical Microbiology, Medical University of Vienna, Vienna, Austria,3 Multimetrix GmbH, Heidelberg, Germany,4 Department of Internal Medicine Hemato-Oncology, University Hospital Brno and Masaryk University Brno, Brno, Czech Republic5
Received 12 August 2008/ Returned for modification 27 September 2008/ Accepted 18 February 2009
In immunocompromised patients suffering from invasive fungal infection, rapid identification of the fungal species is a prerequisite for selection of the most appropriate antifungal treatment. We present an assay permitting reliable identification of a wide range of clinically relevant fungal pathogens based on the high-throughput Luminex microbead hybridization technology. The internal transcribed spacer (ITS2) region, which is highly variable among genomes of individual fungal species, was used to generate oligonucleotide hybridization probes for specific identification. The spectrum of pathogenic fungi covered by the assay includes the most commonly occurring species of the genera Aspergillus and Candida and a number of important emerging fungi, such as Cryptococcus, Fusarium, Trichosporon, Mucor, Rhizopus, Penicillium, Absidia, and Acremonium. Up to three different probes are employed for the detection of each fungal species. The redundancy in the design of the assay should ensure unambiguous fungus identification even in the presence of mutations in individual target regions. The current set of hybridization oligonucleotides includes 75 species- and genus-specific probes which had been carefully tested for specificity by repeated analysis of multiple reference strains. To provide adequate sensitivity for clinical application, the assay includes amplification of the ITS2 region by a seminested PCR approach prior to hybridization of the amplicons to the probe panel using the Luminex technology. A variety of fungal pathogens were successfully identified in clinical specimens that included peripheral blood, samples from biopsies of pulmonary infiltrations, and bronchotracheal secretions derived from patients with documented invasive fungal infections. Our observations demonstrate that the Luminex-based technology presented permits rapid and reliable identification of fungal species and may therefore be instrumental in routine clinical diagnostics.
* Corresponding author. Mailing address: CCRI, Kinderspitalgasse 6, A-1090 Vienna, Austria. Phone: 43-0-1-40470 489. Fax: 43-0-1-40470 437. E-mail: Thomas.Lion{at}ccri.at
Published ahead of print on 25 February 2009.
Supplemental material for this article may be found at http://jcm.asm.org/.
Journal of Clinical Microbiology, April 2009, p. 1063-1073, Vol. 47, No. 4
0095-1137/09/$08.00+0 doi:10.1128/JCM.01558-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»