Molecular differentiation of Bifidobacterium species with amplified ribosomal DNA restriction analysis and alignment of short regions of the ldh gene.
The differentiation of Bifidobacterium species was performed with specific primers using the PCR technique, the amplified ribosomal DNA restriction analysis (ARDRA) technique based on reports on the sequence of the 16S rRNA gene and speciation based on a short region of the ldh gene. Four specific primer sets were developed for each of the Bifidobacterium species, B. animalis, B. infantis and B. longum. The use of the ARDRA method made it possible to discriminate between B. infantis, B. longum and B. animalis with the combination of BamHI, TaqI and Sau3AI restriction enzymes. The ldh gene sequences of 309-312 bp were determined for 19 Bifidobacterium strains. Alignment of these short regions of the ldh gene confirmed that it is possible to distinguish between B. longum and B. infantis but not between B. lactis and B. animalis.
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Food Research and Development Centre, Agriculture and Agri-Food Canada, 3600 Casavant Boulevard West, J2S 8E3, St. Hyacinthe, Que., Canada. royd@em.agr.ca
Sirois
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D000595:Amino Acid Sequence; D001644:Bifidobacterium / Q000145:classification* / Q000235:genetics; D017931:DNA Primers; D004262:DNA Restriction Enzymes; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics*; D005798:Genes, Bacterial; D020459:Genes, rRNA / Q000235:genetics; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D007770:L-Lactate Dehydrogenase; D004262:DNA Restriction Enzymes