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Appl. Environ. Microbiol., 03 1996, 998-1003, Vol 62, No. 3
Copyright © 1996, American Society for Microbiology

Detection of Dekkera-Brettanomyces strains in sherry by a nested PCR method

JI Ibeas, I Lozano, F Perdigones and J Jimenez
Unidad de Genetica, Facultad de Ciencias, Universidad de Malaga, Campus Universitario de Teatinos, Spain.

Brettanomyces sp. and its ascosporogenous sexual state, Dekkera sp., have been well documented as spoilage microorganisms, usually associated with barrel-aged red wines. In this report, we describe the genetic characterization, on the basis of DNA content per cell, electrophoretic karyotyping, and mitochondrial DNA restriction patterns, of a Dekkera yeast strain isolated from sherries and of a number of other Brettanomyces and Dekkera strains. By using a genomic DNA fragment of the isolated Dekkera strain, we developed a two-step PCR method which directs the specific amplification of target DNA from this strain and from other Brettanomyces-Dekkera strains. The method efficiently amplified the target DNA from intact cells, obviating DNA isolation, and yielded a detection limit of fewer than 10 yeast cells in contaminated samples of sherry.

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