Direct detection of Brettanomyces bruxellensis in wine by PCR targeting the vinylphenol reductase gene

Brettanomyces bruxellensis is among the main spoilage yeasts in wine usually found in oak barrels. As routine, laboratories at wineries use selective-differential culture media to detect this yeast. Nevertheless, it is widely recognized that other microbial species can grow on these media, getting false positive signals. In this work, we have developed a conventional PCR method based on the vinylphenol reductase gene (VPR1) of B. bruxellensis applied to wines after a polyvinylpyrrolidone-based pre-treatment (7% w/v) and that allowed us to reach a low detection limit, up to 102 UFC/mL of wine. The procedure was conceived without time-consuming DNA extraction steps, simplifying the methodology and making it easy for applying in wineries