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Applied and Environmental Microbiology, February 2001, p. 938-941, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.938-941.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of Dekkera bruxellensis (Brettanomyces) from Wine by Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes

Henrik Stender,1,* Cletus Kurtzman,2 Jens J. Hyldig-Nielsen,1 Ditte Sørensen,1 Adam Broomer,1 Kenneth Oliveira,1 Heather Perry-O'Keefe,1 Andrew Sage,3 Barbara Young,3 and James Coull1

Boston Probes, Inc., Bedford, Massachusetts 017301; Microbial Properties Research Unit, National Center for Agricultural Utilization Research, USDA Agricultural Research Service, Peoria, Illinois 616042; and Millipore Corporation, Bedford, Massachusetts 017303

Received 28 August 2000/Accepted 2 November 2000

A new fluorescence in situ hybridization method using peptide nucleic acid (PNA) probes for identification of Brettanomyces is described. The test is based on fluorescein-labeled PNA probes targeting a species-specific sequence of the rRNA of Dekkera bruxellensis. The PNA probes were applied to smears of colonies, and results were interpreted by fluorescence microscopy. The results obtained from testing 127 different yeast strains, including 78 Brettanomyces isolates from wine, show that the spoilage organism Brettanomyces belongs to the species D. bruxellensis and that the new method is able to identify Brettanomyces (D. bruxellensis) with 100% sensitivity and 100% specificity.

* Corresponding author. Mailing address: Boston Probes, Inc., 75E Wiggins Ave., Bedford, MA 01730. Phone: (781) 271-1100. Fax: (781) 276-4931. E-mail: HStender{at}BostonProbes.com.

Applied and Environmental Microbiology, February 2001, p. 938-941, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.938-941.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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