title: Lactobacillus species identification by amplified ribosomal 16S-23S rRNA restriction fragment length polymorphism analysis.
abstract: Lactic acid bacteria strains are commonly used for animal and human consumption due to their probiotic properties. One of the major genera used is Lactobacillus, a highly diverse genus comprised of several closely related species. The selection of new strains for probiotic use, especially strains of Lactobacillus, is the focus of several research groups. Accurate identification to species level is fundamental for research on new strains, as well as for safety assessment and quality assurance. The 16S-23S internal transcribed spacer (ITS-1) is a deeply homologous region among prokaryotes that is commonly used for identification to the species level because it is able to acquire and accumulate mutations without compromising general bacterial metabolism. In the present study, 16S-23S ITS regions of 45 Lactobacillus species (48 strains) were amplified and subjected to independent enzymatic digestions, using 12 restriction enzymes that recognise six-base sequences. Twenty-nine species showed unique restriction patterns, and could therefore be precisely identified solely by this assay (64%). This approach proved to be reproducible, allowing us to establish simplified restriction patterns for each evaluated species. The restriction patterns of each species were similar among homologous strains, and to a large extent reflected phylogenetic relationships based on 16S rRNA sequences, demonstrating the promising nature of this region for evolutionary studies.
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: S H C
authors.initials: SH
authors.lastname: Sandes
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: L B
authors.initials: LB
authors.lastname: Alvin
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: B C
authors.initials: BC
authors.lastname: Silva
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: D F
authors.initials: DF
authors.lastname: Zanirati
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: L R C
authors.initials: LR
authors.lastname: Jung
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Nicoli
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: E
authors.initials: E
authors.lastname: Neumann
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Av. Antônio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil.
authors.forename: A C
authors.initials: AC
authors.lastname: Nunes
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S
keywords: 16S-23S rRNA intergenic region; ARDRA; Lactobacillus; species-level identification