Specific Detection of Bifidobacterium Strains in a Pharmaceutical Probiotic Product and in Human Feces by Polymerase Chain Reaction

• Published in: Systematic and applied microbiology Vol. 23; no. 3; pp. 391 - 399
• Main Authors: Brigidi, Patrizia, Vitali, Beatrice, Swennen, Erwin, Altomare, Luigi, Rossi, Maddalena, Matteuzzi, Diego
• Format: Journal Article
• Language: English
• Published: Jena Elsevier GmbH 01.10.2000; Elsevier
• Subjects: 16S-23S rDNA; Bacterial Typing Techniques; Bacteriology; Bifidobacterium; Bifidobacterium - genetics; Bifidobacterium - isolation & purification; Bifidobacterium infantis; Bifidobacterium longum; Biological and medical sciences; Colitis, Ulcerative - microbiology; DNA Primers; DNA, Ribosomal; Feces - microbiology; Fundamental and applied biological sciences. Psychology; Genetics; Human feces; Humans; Identification; Industrial Microbiology; Microbiology; Molecular Sequence Data; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; PCR primers; Polymerase Chain Reaction - methods; Pouchitis - microbiology; Probiotics; RNA, Ribosomal, 16S; RNA, Ribosomal, 23S; Sequence Analysis, DNA; Species Specificity; PCR primers; Probiotics; Human feces; Identification; 16S-23S rDNA; Bifidobacterium; Human; Probiotic; Actinomycetaceae; New product; Ribosomal DNA; Primer; Bifidobacterium longum; Strain specificity; Strain; Actinomycetales; Polymerase chain reaction; 16S-RNA; Bacteria; Actinomycetes; Feces; Detection; By product
• ISSN: 0723-2020; 1618-0984
• DOI: 10.1016/S0723-2020(00)80070-3

For PCR specific detection of the strains Bifidobacterium longum Y 10, B. infantis Y 1 and B. breve Y 8 used in a new probiotic product (VSL-3), strains-specific rDNA primers have been developed. Spacer regions between the 16S and 23S rRNA genes (ITS) of the three strains were amplified by PCR with conserved primers and the nucleotide sequence of these ITSs were determined. On the basis of their comparison with the rDNA sequences retrieved from GenBank, we designed new primers which specifically recognize the species B. breve and the two strains B. infantis Y 1 and B. breve Y 8. Specificity of these primers was confirmed through the analysis of 60 bifidobacteria strains belonging to the more representative human species. The feasibility of this PCR method was investigated in commercial VSL-3 product and fecal samples collected from 4 patients affected by inflammatory bowel deseases and two healthy subjects before and after the VSL-3 administration. By PCR analysis of different VSL-3 commercial batches we were successful in differentiating and quantifying the strains B. longum Y 10, B. infantis Y 1 and B. breve Y 8. B. infantis Y 1 and B. breve Y 8 could be detected at high concentration in fecal specimens of both patients and subjects treated with the probiotic preparation, showing a different colonization behaviour. Seven days after the VSL-3 treatment suspension, no patients and subjects harbored B. infantis Y 1 and B. breve Y 8, indicating a transient presence of these exogenous strains.