title: Multiplex PCR using 16S rRNA gene-targeted primers for the identification of bifidobacteria from human origin.
abstract: Three multiplex polymerase chain reactions (PCRs) targeted on Bifidobacterium and related species were designed to identify human species. The selected primers yielded amplified products of various sizes, each specific for a species. Three to four pairs were gathered in one PCR reaction and their specificity under multiplex conditions was confirmed using DNA from 26 reference strains. Using this technique on unidentified faecal strains, B. bifidum, B. longum and B. breve species were commonly recovered in infants while B. adolescentis, B. catenulatum/B. pseudocatenulatum continuum and B. longum species were predominant in adults. Thus, a single PCR can provide the assignment of a strain to one these species, reducing the number of PCR reactions and hands-on time for the identification of human isolates of bifidobacteria. Moreover, this technique is also applicable for the in situ detection of bifidobacteria in DNA extracts from human stools.
authors.affiliation: Faculté de Pharmacie, 1, rue des Louvels, 80037 Amiens Cedex 1, France. catherine.mullie@sa.u-picardie.fr
authors.forename: Catherine
authors.initials: C
authors.lastname: Mullié
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Odou
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Singer
authors.forename: Marie-Bénédicte
authors.initials: MB
authors.lastname: Romond
authors.forename: Daniel
authors.initials: D
authors.lastname: Izard
mesh_terms: D039941:Bifidobacteriales Infections / Q000382:microbiology*; D001644:Bifidobacterium / Q000145:classification* / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D005243:Feces / Q000382:microbiology; D006801:Humans; D007223:Infant; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S