Comparison of multiplex PCRs, (GTG)5-PCR and automated ribotyping for identification of Lactobacillus spp. isolated from children's intestinal tissues.

• Authors: ŠVEC Pavel; DRÁB Vladimír; TESHIM Andrea; VOLNÁ Lucie; ŠEDO Ondrej; ZDRÁHAL Zbyněk; SEDLÁČEK Ivo
• Year of publication: 2011
• Type: Conference abstract
• MU Faculty or unit: Faculty of Science
• Description: The genus Lactobacillus represents taxonomically diverse group of organisms inhabiting a variety of different environments including the human body. Lactobacilli are generally recognised as most important agents playing beneficial role in balancing microflora of vagina and gastrointestinal tract. This study deals with a group of Lactobacillus strains isolated from the biopsy samples retrieved from children's intestinal tissues. All strains were identified using the Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (Ultraflex III instrument, Bruker Daltonik) and assigned as L. casei/paracasei (11 strains), L. rhamnosus (7), L. mucosae (5), L. salivarius (3), L. gasseri (2), L. plantarum (2), and L. oris (1). Investigated group was further subjected to the multiplex PCR assays based on the integrated sequences of 16S and 23S rRNA genes, (GTG)5-PCR fingerprinting performed by the DiversiLab system (bioMérieux) and automated ribotyping with the RiboPrinter identification system (DuPont Qualicon). Multiplex PCRs identified correctly all L. plantarum, L. rhamnosus and L. salivarius strains, ten L. casei/paracasei, and one L. gasseri strain. Similar results were obtained using (GTG)5-PCR fingerprinting which assigned correctly all L. casei/paracasei, L. gasseri, L. oris, L. rhamnosus and L. salivarius strains. Automatic identification procedure performed by the RiboPrinter system identified only L. casei/paracasei strains, two L. salivarius and one L. gasseri. In conclusion, multiplex PCR assays and (GTG)5-PCR fingerprinting were revealed as good tools for the identification of the investigated intestinal lactobacilli. In contrast, the successfulness of the automatic identification procedure performed by the RiboPrinter system was low. This work was supported by grants 2B08068, LC06034, MSM0021622415 and MSM0021622416 from the Ministry of Education, Youth and Sports of the Czech Republic.

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