Contaminating Brettanomyces/Dekkera yeast are normally found in barrel aged wines. These yeast form potential spoilage compounds such as 4-ethyl-phenol and 4-ethyl-guaiacol. Studies on molecular identification of these potential spoilage yeast were based on the polymorphism of the rRNA and comparative analysis of two chromosome DNA regions situated between the genes encoding for their rRNA. This type of analysis was demonstrated on different strains of Brettanomyces/Dekkera isolated from wine. This study showed that in the internal transcribed spacer (ITS) regions ITS1 and ITS2 are sequentially diverse enough to obtain yeast species identification. PCR techniques using 4 specific primers were tested for their specificity and their reproducibility to detect Brettanomyces/Dekkera. The results of this study showed that by using a combination of these 4 specific primers, a fast and reliable way to identify Brettanomyces/Dekkera presence in wine is possible.