Open AccessRapid Identification and Enumeration of Saccharomyces cerevisiae Cells in Wine by Real-Time PCR
Author(s) -
Patricia Martorell,
Amparo Querol,
M. Teresa Fernández-Espinar
Publication year - 2005
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.71.11.6823-6830.2005
Subject(s) - wine , saccharomyces cerevisiae , yeast , food spoilage , biology , enumeration , saccharomyces , food science , yeast in winemaking , biochemistry , genetics , mathematics , bacteria , combinatorics
Despite the beneficial role ofSaccharomyces cerevisiae in the food industry for food and beverage production, it is able to cause spoilage in wines. We have developed a real-time PCR method to directly detect and quantify this yeast species in wine samples to provide winemakers with a rapid and sensitive method to detect and prevent wine spoilage. Specific primers were designed forS. cerevisiae using the sequence information obtained from a cloned random amplified polymorphic DNA band that differentiatedS. cerevisiae from its sibling speciesSaccharomyces bayanus ,Saccharomyces pastorianus , andSaccharomyces paradoxus . The specificity of the primers was demonstrated for typical wine spoilage yeast species. The method was useful for estimating the level ofS. cerevisiae directly in sweet wines and red wines without preenrichment when yeast is present in concentrations as low as 3.8 and 5 CFU per ml. This detection limit is in the same order as that obtained from glucose-peptone-yeast growth medium (GPY). Moreover, it was possible to quantifyS. cerevisiae in artificially contaminated samples accurately. Limits for accurate quantification in wine were established, from 3.8 × 105 to 3.8 CFU/ml in sweet wine and from 5 × 106 to 50 CFU/ml in red wine.