Detection and identification of Brettanomyces/Dekkera sp. yeasts with a loop-mediated isothermal amplification method.
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.
Hayashi
Nobuyuki
N
Research Laboratories for Brewing, Kirin Brewery Co Ltd, 1-17-1 Namamugi, Yokohama, Japan. n-hayashi@kirin.co.jp
Arai
Ritsuko
R
Tada
Setsuzo
S
Taguchi
Hiroshi
H
Ogawa
Yutaka
Y
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
D017931:DNA Primers; D004271:DNA, Fungal; D012335:RNA, Ribosomal