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Journal of Clinical Microbiology, November 2001, p. 4138-4141, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4138-4141.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Differentiation of Candida albicans and Candida dubliniensis by Fluorescent In Situ Hybridization with Peptide Nucleic Acid Probes

Kenneth Oliveira,1 Gerhard Haase,2 Cletus Kurtzman,3 Jens Jørgen Hyldig-Nielsen,1 and Henrik Stender1,*

Boston Probes, Bedford, Massachusetts1; Institute of Medical Microbiology, University Hospital RWTH Aachen, Aachen, Germany2; and Microbial Properties Research Unit National Center for Agricultural Utilization Research, USDA Agricultural Research Service, Peoria, Illinois3

Received 25 June 2001/Returned for modification 6 August 2001/Accepted 12 August 2001

The recent discovery of Candida dubliniensis as a separate species that traditionally has been identified as Candida albicans has led to the development of a variety of biochemical and molecular methods for the differentiation of these two pathogenic yeasts. rRNA sequences are well-established phylogenetic markers, and probes targeting species-specific rRNA sequences have been used in diagnostic assays for the detection and identification of microorganisms. Peptide nucleic acid (PNA) is a DNA mimic with improved hybridization characteristics, and the neutral backbone of PNA probes offers significant advantages in whole-cell in situ hybridization assays. In this study, we developed PNA probes targeting the rRNAs of C. albicans and C. dubliniensis and applied them to a fluorescence in situ hybridization method (PNA FISH) for differentiation between C. albicans and C. dubliniensis. Liquid cultures were smeared onto microscope slides, heat fixed, and then hybridized for 30 min. Unhybridized PNA probe was removed by washing, and smears were examined by fluorescence microscopy. Evaluation of the PNA FISH method using smears of 79 C. dubliniensis and 70 C. albicans strains showed 100% sensitivity and 100% specificity for both PNA probes. We concluded that PNA FISH is a powerful tool for the differentiation of C. albicans and C. dubliniensis.

* Corresponding author. Mailing address: Boston Probes, 15 DeAngelo Dr., Bedford, MA 01730. Phone: (781) 271-1100, ext. 291. Fax: (781) 276-4931. E-mail: hstender{at}BostonProbes.com.

Journal of Clinical Microbiology, November 2001, p. 4138-4141, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4138-4141.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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