Related Experiment Videos
Specific detection of a probiotic Lactobacillus strain in faecal samples by using multiplex PCR
1Istituto di Microbiologia, Facoltà di Agraria, Università Cattolica del Sacro Cuore, Piacenza, Italy.
FEMS Microbiology Letters
|February 18, 1998
Summary
This study developed a rapid method to identify Lactobacillus gasseri 4B2 using its aggregation-promoting factor (APF) gene. The technique allows direct bacterial cell analysis, simplifying strain identification in various settings.
Area of Science:
- Microbiology
- Molecular Biology
- Probiotics
Background:
- Lactobacillus gasseri 4B2 exhibits a strong autoaggregating phenotype.
- This autoaggregation is mediated by the secreted protein, aggregation-promoting factor (APF).
Purpose of the Study:
- To develop a rapid and reliable method for identifying Lactobacillus gasseri 4B2.
- To enable direct identification of the strain without DNA extraction.
Main Methods:
- Developed two primer pairs for simultaneous amplification of the APF gene and the 16S rRNA gene.
- Tested the protocol on single and mixed Lactobacillus cultures.
- Validated the method using whole bacterial cells directly added to PCR reactions.
- Assessed in vivo applicability by feeding L. gasseri 4B2 to mice and analyzing fecal samples.
Main Results:
- The developed PCR protocol successfully amplified specific fragments of the APF and 16S rRNA genes.
- The method demonstrated specificity in identifying L. gasseri 4B2 in both single and mixed cultures.
- Direct addition of whole bacterial cells to PCR reactions yielded reliable amplification.
- The methodology proved suitable for in vivo studies, allowing analysis of fecal samples.
Conclusions:
- A fast and reliable method for identifying Lactobacillus gasseri 4B2 has been established.
- This technique eliminates the need for DNA extraction, simplifying the identification process.
- The method is applicable for both in vitro and in vivo studies, enhancing probiotic research.