Dot blot and PCR for Brettanomyces bruxellensis detection in red wine

- Specific primers and a DNA dig-oxygenin labelled probe were used in PCR and dot blot.
- PCR sensitivity with DekITS-BruxITR was 10 pg μl−1 using pure Brettanomyces bruxellensis DNA.
- DNA digoxigenin labelled probe detected 10 CFU μl−1B. bruxellensis in wine samples.
- PCR and Dot blot are useful for B. bruxellensis colony confirmation and for wine analyses.

The capability to identify in a short time Brettanomyces bruxellensis, the yeast causing wine spoilage, is an important result both to increase quality of the products and to reduce economic losses in wineries. The development of a quick and specific method to detect B. bruxellensis in wine samples is a good tool for routine controls in wineries. Specific primers (DekITS and BruxITR) and a specific digoxygenin labelled probe were designed to be used in a PCR protocol and in a Dot blot method to detect B. bruxellensis in wine samples and/or to confirm isolates from wine samples. A molecular method, not requiring colony growth, is helpful for winemakers alike to quickly obtain information about the presence of Brettanomyces in their products. The results of this work indicate that the dot blot method is sensitive and fits this goal showing a great potential for its simple applicability. Both methods PCR and dot blot were useful for both a direct detection and a species specific confirmation of the isolates.