Development of a genus-specific T-RFLP technique for differentiating Lactobacillus community

Abstract:

[Objective] To develop a Lactobacillus genus-specific T-RFLP technique for differentiating 14 Lactobacillus strains. [Methods] A Lactobacillus genus-specific primer (LAB-rev) was firstly designed based on the sequence variation of bacterial 16S-23S space region, which was then labeled with 6-Carboxyfluorescein (6-FAM), followed by PCR with a 16S universal primer (7f). The PCR amplicon was then digested by restriction enzyme Hae III and HhaI before terminal sequencing by AB3730 Gene Scan to achieve T-RFLP profiles. [Results] Results showed that this technique is more specific than the traditional T-RFLP using universal primers, which can be used to detect and quantitate Lactobacillus of 14 different species in this study. [Conclusion] This technique has a great potential of application in investigating lactobacillus communities, evaluating the probiotic function of functional food, dairy drinks and medicine on intestinal micro-ecological environment.