Quantitative assessment of faecal bifidobacterial populations by real-time PCR using lanthanide probes

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Abstract
Aim: To develop real-time quantitative PCR methods, based on the use of\ud probes labelled with a stable fluorescent lanthanide chelate, for the quantification\ud of different human faecal bifidobacterial populations.\ud Methods and Results: The designed quantitative PCR assays were found to\ud be specific for the corresponding Bifidobacterium species or groups (Bifidobacterium\ud longum group, Bifidobacterium catenulatum group, Bifidobacterium\ud adolescentis, Bifidobacterium breve, Bifidobacterium angulatum, Bifidobacterium\ud bifidum and Bifidobacterium dentium). The detection limits of the methodologies\ud used ranged between 2 · 105 and 9 · 103 cells g)1 of faeces. The applicability\ud of the developed assays was tested by analysing 20 human faecal samples.\ud Bif. longum group was found to be the qualitatively and quantitatively predominant\ud bifidobacterial group.\ud Conclusions: The real-time PCR procedures developed here are specific, accurate,\ud rapid and easy methods for the quantification of Bifidobacterium groups\ud or species in human faecal samples.\ud Significance and Impact of the Study: The developed procedures will facilitate\ud rapid and objective counting of large numbers of samples increasing our knowledge\ud on the role of gut bifidobacterial microbiota in health and disease. This\ud will contribute to the efficient use of intestinal bacterial assays in research, food\ud and pharmaceutical development as well as in the assessment of dietary management\ud of diseases.Financial support for this study was obtained from the Academy of Finland and by an Unrestricted Nutrition Research Grant from Bristol Myers Squibb Foundation.Peer reviewe

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