title: Rapid identification of the genus Dekkera/Brettanomyces, the Dekkera subgroup and all individual species.
abstract: The genus Dekkera/Brettanomyces comprises five described species: Dekkera bruxellensis, D. anomala, Brettanomyces custersianus, B. naardenensis and B. nanus. Some of them, especially D. bruxellensis, are important spoilage organisms, particularly in the wine and beverage industries. Because of their economic importance many different methods have been developed to identify members of the genus in general and D. bruxellensis in particular. These methods vary in their rapidity, complexity and cost but, partly because of confidentiality issues, it is unclear which methods are used, or how widely, in the relevant industries. Building on previous work with the genera Saccharomyces and Zygosaccharomyces, a suite of eight PCR primer pairs has been designed either on the D1-D2 region of the 26S rRNA gene or translation elongation factor TEF1-α. These primers can specifically identify the genus as a whole, only Dekkera species, each one of the five recognised species as well as a significant subgroup of D. bruxellensis represented by NCYC 3426. Multiplexing has also been tried and it has been shown to be possible with some combinations of genus or Dekkera-level and species-specific primers. Using direct colony PCR amplification followed by gel electrophoresis, a clear positive result can be obtained in less than 3h, thus providing a quick, reliable and inexpensive way to identify target species.
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK. Electronic address: Michelle.Hulin@emr.ac.uk.
authors.forename: M
authors.initials: M
authors.lastname: Hulin
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK. Electronic address: elizabethharrison77@gmail.com.
authors.forename: E
authors.initials: E
authors.lastname: Harrison
authors.affiliation: School of Life Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, UK. Electronic address: Malcolm.Stratford@mologic.co.uk.
authors.forename: M
authors.initials: M
authors.lastname: Stratford
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK. Electronic address: bssaew@bath.ac.uk.
authors.forename: A E
authors.initials: AE
authors.lastname: Wheals
mesh_terms: D055168:Brettanomyces / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D055167:Dekkera / Q000235:genetics / Q000302:isolation & purification*; D015977:Eukaryotic Initiation Factor-1 / Q000235:genetics; D005516:Food Microbiology / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000235:genetics; D013045:Species Specificity; D014920:Wine / Q000382:microbiology*; D020068:Zygosaccharomyces / Q000235:genetics
chemical_list: D017931:DNA Primers; D015977:Eukaryotic Initiation Factor-1; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S; C026022:eukaryotic peptide initiation factor-1A
keywords: Colony PCR; Dekkera/Brettanomyces; Rapid identification