Identification of Dekkera bruxellensis (Brettanomyces) from wine by fluorescence in situ hybridization using peptide nucleic acid probes.
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.
Stender
H
H
Boston Probes, Inc., Bedford, Massachusetts 01730, USA. HStender@BostonProbes.com
Kurtzman
C
C
Hyldig-Nielsen
J J
JJ
Sørensen
D
D
Broomer
A
A
Oliveira
K
K
Perry-O'Keefe
H
H
Sage
A
A
Young
B
B
Coull
J
J
D001483:Base Sequence; D004275:DNA, Ribosomal / Q000032:analysis; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D015341:Nucleic Acid Probes / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000235:genetics*; D012335:RNA, Ribosomal / Q000235:genetics; D013045:Species Specificity; D014920:Wine / Q000382:microbiology*; D015003:Yeasts / Q000145:classification* / Q000235:genetics
D004275:DNA, Ribosomal; D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D012335:RNA, Ribosomal