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?lactis. Using the xfp DNA fragment of B.?lactis as standard, the qPCR assay detection range was between 0��0625?fg DNA (corresponding to 260 B.?lactis cells) and 6250?fg DNA (corresponding to 2��6?��?107B.?lactis cells). This assay for bifidobacteria was linear for CT values from 16 to 32. The assay with the primer Eub340?F and Eub781 R showed a detection range from 62��5?fg to 625?pg DNA and was linear for CT values between 16 and 31 corresponding to 1��5?��?104 and 1��5?��?108 copies of the 16S rRNA gene respectively. Both standard curves (Fig.?2) used to quantify bifidobacteria and total bacteria showed a high linear correlation (R2) coefficient of 0��997 and 0��999 for the xfp gene and pLME21 plasmid respectively. The slopes of the standard lines for xfp and total bacteria were very similar with ?3��627 and ?3��637 for calculated amplification http://www.selleckchem.com/products/bmn-673.html efficiencies of 94��3 and 94��2% respectively. Only small matrix-dependent effects were observed when comparing the mean xfp gene copy number in spiked, autoclaved faecal samples and pure cultures of B.?longum. After subtracting the background bifidobacterial cell numbers measured in autoclaved faeces (2��5?��?103?xfp?copies?g?1), the mean recovery rate of bifidobacterial DNA in spiked faecal samples was 78��2?��?6��6% (n?=?3), regardless of the initial concentration of B.?longum. The sensitivity of the qPCR method for Bifidobacterium spp. was assessed by analysing http://www.selleckchem.com/products/pexidartinib-plx3397.html serial 10-fold dilutions of B.?longum spiked in autoclaved faeces. The detection limits for the bifidobacteria and 16S rRNA gene (total bacteria) assay were 2��5?��?103 and 2��5?��?105?bacteria?g?1 of faeces respectively. Significantly higher total bacterial concentrations (P? http://www.selleck.cn/products/nlg919.html of 9��3?��?0��3 log10?cell?numbers?g?1) and children (mean of 9��2?��?0��3 log10?cell?numbers?g?1). In contrast, the FF infant harboured a lower concentration of bifidobacteria (9��6?log10?cell?numbers?g?1). The proportion of bifidobacteria (MS-Bif164) among total bacteria (MS-EUB338) ranged from 0��9% to 3��1% in adults, from 1��4% to 6��7% in children, and was 59% in the BF infant and only 9% in the FF infant. This microscopic detection method showed low intra-assay coefficients of variation (CVassay, n?=?3) ranging from 0��2% to 5��6% in MS-DAPI, MS-EUB338 and MS-Bif164 for all age groups.