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PMID: 8837422 Published · ppublish English Clinical Trial Journal Article Randomized Controlled Trial

Specific detection and analysis of a probiotic Bifidobacterium strain in infant feces.

Applied and environmental microbiology ·Vol. 62 ·No. 10 ·1996-10-00 ·Pages 3668-72

Kok RG, de Waal A, Schut F, Welling GW, Weenk G, Hellingwerf KJ

Abstract

For specific detection of the probiotic Bifidobacterium sp. strain LW420 in infant feces and for rapid quality control of this strain in culture, three strain-specific 16S rRNA gene-targeted primers have been developed. These primers allow specific detection of the organism via PCR. Specificity of the primers was determined in DNA samples isolated from single-strain and mixed cultures of bifidobacteria and in heterogenous fecal samples. The feasibility of this method for use in specific detection of probiotic strains was investigated through addition of Bifidobacterium sp. strain LW420 to infant instant milk formula (IMF) and PCR analyses of bacterial DNA isolated from feces of 17 newborn IMF-fed infants. In feces of all nine babies that had been fed with the probiotic IMF, the strain-specific PCR signal could be detected. No signal was found in feces of the eight infants that had been fed with a nonprobiotic IMF, demonstrating the specificity of the PCR method. All 17 infants developed a major fecal Bifidobacterium population already after 3 days, as determined through genus-specific and strain-specific PCR. Phenotypical screening of Bifidobacterium sp. strain LW420 and analysis of homology of the 16S rRNA gene sequence of this strain with that of other bifidobacteria deposited in databases do not allow positive classification of LW420 among the currently known species of Bifidobacterium.

MeSH Terms
Animals Bifidobacterium/genetics,growth & development,isolation & purification DNA Primers DNA, Ribosomal/genetics Double-Blind Method Feces/microbiology Fermentation Humans Infant Food/microbiology Infant, Newborn Milk Molecular Sequence Data Polymerase Chain Reaction/methods RNA, Ribosomal, 16S/genetics Sensitivity and Specificity Sequence Analysis, DNA Sequence Homology, Nucleic Acid
Chemicals
DNA Primers DNA, Ribosomal RNA, Ribosomal, 16S
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kok R G
Department of Microbiology, E. C. Slater Institute, BioCentrum Amsterdam, The Netherlands.
de Waal A
Schut F
Welling G W
Weenk G
Hellingwerf K J
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
Published
1996-10-00
Pages
3668-72
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC168175
Subset
IM
Databases
GENBANK
X89111
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