Multiplex PCR using 16S rRNA gene-targeted primers for the identification of bifidobacteria from human origin

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 sp...

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Bibliographic Details
Published in:FEMS microbiology letters 2003-05, Vol.222 (1), p.129-136
Main Authors: Mullié, Catherine, Odou, Marie-Françoise, Singer, Elisabeth, Romond, Marie-Bénédicte, Izard, Daniel
Format: Article
Language:English
Subjects:
16S rDNA
adults
Bacteriology
Bifidobacteriales Infections - microbiology
Bifidobacterium
Bifidobacterium - classification
Bifidobacterium - genetics
Biological and medical sciences
DNA
DNA Primers
DNA, Bacterial - analysis
Feces - microbiology
Fundamental and applied biological sciences. Psychology
Human
Humans
Infant
infants
Microbiology
Miscellaneous
Multiplex polymerase chain reaction
Phylogeny
polymerase chain reaction
Polymerase Chain Reaction - methods
ribosomal RNA
RNA, Ribosomal, 16S - genetics
Species identification
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Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(03)00245-3