Valio Ltd, R&D, Helsinki, Finland.
J Appl Microbiol. 2009 Feb;106(2):506-14. doi: 10.1111/j.1365-2672.2008.04018.x.
To develop a strain-specific rapid assay for identification and quantification of Lactobacillus rhamnosus GG in human faecal samples.
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.
Quantitative real-time PCR is a suitable method for strain-specific identification of L. rhamnosus GG in human faecal samples.
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.
开发一种用于鉴定和定量人粪便样本中鼠李糖乳杆菌 GG 的菌株特异性快速检测方法。
分离并测序了鼠李糖乳杆菌 GG 菌株的独特随机扩增多态性 DNA（RAPD）带。基于该序列设计了菌株特异性聚合酶链反应（PCR）引物和探针。通过荧光共振能量转移（FRET）系统的实时 PCR 进行定量。该方法的特异性通过一组已知菌株和人粪便样本的 DNA 进行了测试。该方法对鼠李糖乳杆菌 GG 的分析灵敏度约为每个检测 10 CFU，相当于湿粪便中 10(5) CFU g(-1)。
实时定量 PCR 是一种适合鉴定人粪便样本中鼠李糖乳杆菌 GG 的菌株特异性方法。
鼠李糖乳杆菌 GG 是临床试验中研究最多的益生菌菌株之一，但仍缺乏基于 DNA 的鉴定方法。本研究描述了一种用于鉴定和定量人粪便样本中鼠李糖乳杆菌 GG 的菌株特异性实时 PCR 方法。
