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. 2002 Jun;40(6):2182-6.
doi: 10.1128/JCM.40.6.2182-2186.2002.

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, Gerhard Haase, Deborah Wilson, Geraldine Hall, Cletus Kurtzman, Kenneth Oliveira, Sabina Von Oy, Jens J Hyldig-Nielsen, James Coull, Henrik Stender
Affiliations

Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles

Susan Rigby et al. J Clin Microbiol. 2002 Jun.

Abstract

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 degrees 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.

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Figures

FIG. 1.
FIG. 1.
Images of two routine yeast-positive blood culture smears analyzed by the C. albicans PNA FISH method. The organism in panel A was identified as C. glabrata by standard methods, and the organism in panel B was identified as C. albicans by standard methods. C. albicans cells detected by the C. albicans PNA FISH method appear as bright green fluorescent yeast cells on a reddish smear background, whereas C. glabrata cells are slightly visible as reddish yeast cells infiltrated in the smear.
See this image and copyright information in PMC

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