title: Detection and identification of Brettanomyces/Dekkera sp. yeasts with a loop-mediated isothermal amplification method.
abstract: Primer sets for a loop-mediated isothermal amplification (LAMP) method were developed to specifically identify each of the four Brettanomyces/Dekkera species, Dekkera anomala, Dekkera bruxellensis, Dekkera custersiana and Brettanomyces naardenensis. Each primer set was designed with target sequences in the ITS region of the four species and could specifically amplify the target DNA of isolates from beer, wine and soft drinks. Furthermore, the primer sets differentiated strains of the target species from strains belonging to other species, even within the genus Brettanomyces/Dekkera. Moreover, the LAMP method with these primer sets could detect about 1 x 10(1) cfu/ml of Brettanomyces/Dekkera yeasts from suspensions in distilled water, wine and beer. This LAMP method with primer sets for the identification of Brettanomyces/Dekkera yeasts is advantageous in terms of specificity, sensitivity and ease of operation compared with standard PCR methods.
authors.affiliation: Research Laboratories for Brewing, Kirin Brewery Co Ltd, 1-17-1 Namamugi, Yokohama, Japan. n-hayashi@kirin.co.jp
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Hayashi
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Arai
authors.forename: Setsuzo
authors.initials: S
authors.lastname: Tada
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Taguchi
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Ogawa
mesh_terms: D000434:Alcoholic Beverages / Q000382:microbiology; D001483:Base Sequence; D001628:Beverages / Q000382:microbiology; D002253:Carbonated Beverages / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D005516:Food Microbiology; D005784:Gene Amplification; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D004718:Saccharomycetales / Q000145:classification* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D012335:RNA, Ribosomal