AIMS: To develop a strain-specific rapid assay for identification and quantification of Lactobacillus rhamnosus GG in human faecal samples. METHODS AND RESULTS: A unique random amplified polymorphic DNA (RAPD) band of the L. rhamnosus GG strain was isolated and sequenced. Strain-specific polymerase chain reaction (PCR) primers and probes were designed based on the sequence. Quantification was performed by the real-time PCR using a fluorescent resonance energy transfer (FRET) system. The specificity of the assay was tested with DNA isolated from a set of known strains and human faecal samples. The analytical sensitivity of the method for L. rhamnosus GG was about 10 CFU per assay, which corresponds to 10(5) CFU g(-1) of wet faeces. CONCLUSIONS: Quantitative real-time PCR is a suitable method for strain-specific identification of L. rhamnosus GG in human faecal samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus rhamnosus GG is one of the most studied probiotic strains in clinical trials but still lacks a DNA-based identification method. This study describes a real-time PCR method for strain-specific identification and quantification of L. rhamnosus GG in human faecal samples.
AIMS: To develop a strain-specific rapid assay for identification and quantification of Lactobacillus rhamnosus GG in human faecal samples. METHODS AND RESULTS: A unique random amplified polymorphic DNA (RAPD) band of the L. rhamnosus GG strain was isolated and sequenced. Strain-specific polymerase chain reaction (PCR) primers and probes were designed based on the sequence. Quantification was performed by the real-time PCR using a fluorescent resonance energy transfer (FRET) system. The specificity of the assay was tested with DNA isolated from a set of known strains and human faecal samples. The analytical sensitivity of the method for L. rhamnosus GG was about 10 CFU per assay, which corresponds to 10(5) CFU g(-1) of wet faeces. CONCLUSIONS: Quantitative real-time PCR is a suitable method for strain-specific identification of L. rhamnosus GG in human faecal samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus rhamnosus GG is one of the most studied probiotic strains in clinical trials but still lacks a DNA-based identification method. This study describes a real-time PCR method for strain-specific identification and quantification of L. rhamnosus GG in human faecal samples.
Authors: Christopher D Packey; Michael T Shanahan; Sayeed Manick; Maureen A Bower; Melissa Ellermann; Susan L Tonkonogy; Ian M Carroll; R Balfour Sartor Journal: Gut Microbes Date: 2013-07-24
Authors: François P Douillard; Angela Ribbera; Kun Xiao; Jarmo Ritari; Pia Rasinkangas; Lars Paulin; Airi Palva; Yanling Hao; Willem M de Vos Journal: Appl Environ Microbiol Date: 2016-06-13 Impact factor: 4.792
Authors: Yvonne E M Dommels; Robèr A Kemperman; Yvonne E M P Zebregs; René B Draaisma; Arne Jol; Danielle A W Wolvers; Elaine E Vaughan; Ruud Albers Journal: Appl Environ Microbiol Date: 2009-08-14 Impact factor: 4.792