Genus-specific T-RFLP technique for differentiating Bifidobacteria community in probiotics

Abstract:

[Objective] A Bifidobacteria genus-specific T-RFLP technique was developed to differentiate Bifidobacterium strains in microbial communities. [Methods] A Bifidobacteria genus-specific primer (g-bifid-F), based on the bacterial 16S rRNA gene sequence, was 5′-labeled with hexachlorofluorescein (HEX), followed by PCR with a 16S universal primer (1510r). The PCR amplicon was then digested by restriction enzyme Hae III and Alu I before terminal sequencing to achieve T-RFLP profiles. Moreover, combined with a Lactobacillus genus-specific T-RFLP technique in our lab’s previous study, this technique was used to detect bacterial composition and the compositional dynamic of a commercial probiotic product—Biostime. [Results] This technique was more specific than the traditional T-RFLP using universal primers, which could be used to detect or semi-quantitate different species of Bifidobacteria. [Conclusion] A Bifidobacteria genus-specific T-RFLP technique was well-established, and a combined system of these two T-RFLP techniques was successfully applied to commercial probiotic product detection.