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Journal of Clinical Microbiology, October 2005, p. 4943-4953, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.4943-4953.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of a DNA Microarray for Detection and Identification of Fungal Pathogens Involved in Invasive Mycoses

Dirk M. Leinberger,1 Ulrike Schumacher,2 Ingo B. Autenrieth,2 and Till T. Bachmann1*

Institute of Technical Biochemistry, University of Stuttgart, Stuttgart, Germany,1 Institute of Medical Microbiology and Hygiene, Eberhard-Karls-University, Tübingen, Germany2

Received 15 April 2005/ Returned for modification 15 May 2005/ Accepted 5 July 2005

Invasive fungal infections have emerged as a major cause of morbidity and mortality in immunocompromised patients. Conventional identification of pathogenic fungi in clinical microbiology laboratories is time-consuming and, therefore, often imperfect for the early initiation of an adequate antifungal therapy. We developed a diagnostic microarray for the rapid and simultaneous identification of the 12 most common pathogenic Candida and Aspergillus species. Oligonucleotide probes were designed by exploiting the sequence variations of the internal transcribed spacer (ITS) regions of the rRNA gene cassette to identify Candida albicans, Candida dubliniensis, Candida krusei, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida guilliermondii, Candida lusitaniae, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, and Aspergillus terreus. By using universal fungal primers (ITS1 and ITS4) directed toward conserved regions of the 18S and 28S rRNA genes, respectively, the fungal ITS target regions could be simultaneously amplified and fluorescently labeled. To establish the system, 12 precharacterized fungal strains were analyzed; and the method was validated by using 21 clinical isolates as blinded samples. As the microarray was able to detect and clearly identify the fungal pathogens within 4 h after DNA extraction, this system offers an interesting potential for clinical microbiology laboratories.

* Corresponding author. Mailing address: Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, Stuttgart 70569, Germany. Phone: 49 (0) 711 685 3197. Fax: 49 (0) 711 685 3196. E-mail: Till.Bachmann{at}itb.uni-stuttgart.de.

Journal of Clinical Microbiology, October 2005, p. 4943-4953, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.4943-4953.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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