The molecular detection of brettanomyces wild yeast in raw wines

Abstract

In this work, home-designed primers for detection of Brettanomyces in wine by Sybr Green I-based real-time PCR (Polymerase Chain Reaction) were developed. Primers were tested both in silico and in vitro. The primers were designed using the sequence of Brettanomyces internal transcribed spacer 1, partial sequence; 5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and large subunit ribosomal RNA gene, partial sequence, available in GeneBank. Primer specificity was checked by aligning the primers against the sequences available in GeneBank using BLAST. Primer performance was tested using positive (wine sample infected with Brettanomyces) and negative controls.