Real-time PCR assay for detection and enumeration of Dekkera bruxellensis in wine.
Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.
Phister
Trevor G
TG
Department of Viticulture and Enology, University of California, Davis, California 95616, USA.
Mills
David A
DA
D001483:Base Sequence; D015169:Colony Count, Microbial; D017931:DNA Primers; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005506:Food Contamination; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D004718:Saccharomycetales / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment; D013045:Species Specificity; D014920:Wine / Q000382:microbiology*
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S