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Journal of Clinical Microbiology, June 2002, p. 2182-2186, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2182-2186.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes for Rapid Identification of Candida albicans Directly from Blood Culture Bottles

Susan Rigby,1 Gary W. Procop,2 Gerhard Haase,3 Deborah Wilson,2 Geraldine Hall,2 Cletus Kurtzman,4 Kenneth Oliveira,1 Sabina Von Oy,3 Jens J. Hyldig-Nielsen,1 James Coull,1 and Henrik Stender1*

Applied Biosystems, Bedford, Massachusetts,1 Section of Clinical Microbiology, Cleveland Clinic Foundation, Cleveland, Ohio,2 Institute of Medical Microbiology, University Hospital RWTH Aachen, Aachen, Germany,3 National Center for Agricultural Utilization Research, Peoria, Illinois4

Received 10 December 2001/ Returned for modification 28 January 2002/ Accepted 26 March 2002

A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55°C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management.

* Corresponding author. Mailing address: Applied Biosystems, Bedford, MA 01730. Phone: (781) 280-2845. Fax: (781) 280-2940. E-mail: Henrik.Stender{at}AppliedBiosystems.com.

Journal of Clinical Microbiology, June 2002, p. 2182-2186, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2182-2186.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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