title: A genus-specific PCR method for differentiation between Leuconostoc and Weissella and its application in identification of heterofermentative lactic acid bacteria from coffee fermentation.
abstract: A genus-specific PCR analysis method was developed for a rapid and reliable differentiation between the two heterofermentative lactic acid bacteria genera Leuconostoc and Weissella. Primer sets specific for target regions of the 16S rRNA genes were designed and the specificity of the PCR was evaluated using the type strains of 13 species of Leuconostoc and 11 species of Weissella. In addition, the newly developed genus-specific PCR analysis was applied to characterize 72 lactic acid bacteria (LAB) strains isolated from coffee fermentation and which were presumptively classified as Leuconostoc or Weissella species. Additionally, a total of 34 LAB isolates from various other fermented foods were included. The investigations of these strains were conducted to test the effectiveness of correct characterization of field isolates using the genus-specific PCR approach. The correct assignment to one of these two genera by the application of the genus-specific primers was confirmed by further identifying the strains using repetitive extragenic palindromic-PCR and 16S rRNA gene sequencing.
authors.affiliation: Federal Research Centre for Nutrition and Foods, Institute of Hygiene and Toxicology, Karlsruhe, Germany. ulrich.schillinger@mri.bund.de
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schillinger
authors.forename: Benjamin
authors.initials: B
authors.lastname: Boehringer
authors.forename: Sabrina
authors.initials: S
authors.lastname: Wallbaum
authors.forename: Lily
authors.initials: L
authors.lastname: Caroline
authors.forename: Almaz
authors.initials: A
authors.lastname: Gonfa
authors.forename: Melanie
authors.initials: M
authors.lastname: Huch Née Kostinek
authors.forename: Wilhelm H
authors.initials: WH
authors.lastname: Holzapfel
authors.forename: Charles M A P
authors.initials: CM
authors.lastname: Franz
mesh_terms: D003069:Coffee / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005285:Fermentation; D005516:Food Microbiology; D020459:Genes, rRNA / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D003069:Coffee; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S