Real-time PCR assays for strain-specific quantification of probiotic strains in human faecal samples
| Autor: | Heli Karjalainen, Terhi Ahlroos, Soile Tynkkynen, Eveliina Myllyluoma |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Bifidobacterium breve
biology Propionibacterium freudenreichii ved/biology ved/biology.organism_classification_rank.species food and beverages biology.organism_classification Applied Microbiology and Biotechnology Genome law.invention RAPD Microbiology Probiotic Real-time polymerase chain reaction Lactobacillus rhamnosus law Polymerase chain reaction Food Science |
| Zdroj: | International Dairy Journal. 27:58-64 |
| ISSN: | 0958-6946 |
| Popis: | Strain-specific real-time PCR assays were developed for the probiotic strains Bifidobacterium breve 99, Lactobacillus rhamnosus LC705 and Propionibacterium freudenreichii ssp. shermanii JS, used in combination, for quantification of the strains in human faecal samples. Unique randomly amplified polymorphic DNA (RAPD) sequences or a phage-related sequence of the strains were used as a basis for designing strain-specific primers and probes. The real-time PCR conditions were optimised and the specificities of the assays were confirmed by analysing DNA from related bacterial strains and from faecal samples originating from an intervention study. The assays allowed quantification with a linear range of six orders of magnitude, from 10 1 to 10 7 target genomes per assay. The detection limit was 4 × 10 5 genome copies per g of faeces. The strain-specific quantitative real-time PCR assays developed were suitable for identification and quantification of probiotic strains in human faecal samples in a controlled intervention trial. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect