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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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