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. 2001 Feb;67(2):938-41.
doi: 10.1128/AEM.67.2.938-941.2001.

Identification of Dekkera bruxellensis (Brettanomyces) from wine by fluorescence in situ hybridization using peptide nucleic acid probes

H Stender  1 , C Kurtzman, J J Hyldig-Nielsen, D Sørensen, A Broomer, K Oliveira, H Perry-O'Keefe, A Sage, B Young, J Coull
Affiliations

Identification of Dekkera bruxellensis (Brettanomyces) from wine by fluorescence in situ hybridization using peptide nucleic acid probes

H Stender et al. Appl Environ Microbiol. 2001 Feb.

Abstract

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.

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Figures

FIG. 1
FIG. 1
Alignment of partial yeast D1-D2 26S rDNA sequences for probe selection. The anti-parallel hybridization sequence of the BRE26S14 PNA probe is shown above the alignment. Base differences between the target sequences and other sequences are highlighted.
FIG. 2
FIG. 2
Microscope images of cells from colonies of a negative control (spheroid yeast #1; Beringer) (A) and a Brettanomyces-positive wine sample (Vinquiry, Inc.) (B).
See this image and copyright information in PMC

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