Abstract
A group of 67 Lactobacillus spp. strains containing Lactobacillus casei/paracasei, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus salivarius species isolated from early childhood caries and identified to the species level in a previous study (Švec et al., Folia Microbiol 54:53–58, 2009) was characterized by automated ribotyping performed by the RiboPrinter® microbial characterization system and by randomly amplified polymorphic DNA fingerprinting (RAPD-PCR) with M13 primer to evaluate these techniques for characterization of lactobacilli associated with dental caries. Ribotyping revealed 55 riboprints among the analysed group. The automatic identification process performed by the RiboPrinter system identified 18 strains to the species level, however cluster analysis divided obtained ribotype patterns into individual clusters mostly corresponding to the species assignment of particular strains. RAPD-PCR fingerprints revealed by the individual Lactobacillus spp. showed higher variability than the ribotype patterns and the fingerprint profiles generated by the analysed species were distributed among one to four clusters. In conclusion, ribotyping is shown to be more convenient for the identification purposes while RAPD-PCR fingerprinting results indicate this method is a better tool for typing of Lactobacillus spp. strains occurring in dental caries.
Similar content being viewed by others
References
Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE (2005) Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 43:5721–5732
Andrighetto C, Zampese L, Lombardi A (2001) RAPD-PCR characterization of lactobacilli isolated from artisanal meat plants and traditional fermented sausages of Veneto region (Italy). Lett Appl Microbiol 33:26–30
Ben Amor K, Vaughan EE, de Vos WM (2007) Advanced molecular tools for the identification of lactic acid bacteria. J Nutr 137:741S–747S
Bingen EH, Denamur E, Elion J (1994) Use of ribotyping in epidemiologic surveillance of nosocomial outbreaks. Clin Microbiol Rev 7:311–327
Bouchet V, Huot H, Goldstein R (2008) Molecular genetic basis of ribotyping. Clin Microbiol Rev 21:262–273
Brisse S, Fussing V, Ridwan B, Verhoef J, Willems RJL (2002) Automated ribotyping of vancomycin-resistant Enterococcus faecium isolates. J Clin Microbiol 40:1977–1984
Caetano-Anolles G (1993) Amplifying DNA with arbitrary oligonucleotide primers. PCR Methods Appl 3:85–94
Dellaglio F, Dicks LMT, Dutoit M, Torriani S (1991) Designation of ATCC 334 in place of ATCC 393 (NCDO 161) as the neotype strain of Lactobacillus casei subsp. casei and rejection of the name Lactobacillus paracasei. Int J Syst Bacteriol 41:340–342
Dellaglio F, Felis GE, Torriani S et al (2002) The status of the species Lactobacillus casei (Orla-Jensen 1916) Hansen and Lessel 1971 and Lactobacillus paracasei Collins et al. 1989. Request for an opinion. Int J Syst Evol Microbiol 52:285–287
Dicks LMT, Du Plessis EM, Dellaglio F, Lauer E (1996) Reclassification of Lactobacillus casei subsp. casei ATCC 393 and Lactobacillus rhamnosus ATCC 15820 as Lactobacillus zeae nom. rev., designation of ATCC 334 as the neotype of L. casei subsp. casei, and rejection of the Lactobacillus paracasei. Int J Syst Bacteriol 46:337–340
Gatti M, Lazzi C, Rossetti L, Mucchetti G, Neviani E (2003) Biodiversity in Lactobacillus helveticus strains present in natural whey starter used for Parmigiano Reggiano cheese. J Appl Microbiol 95:463–470
Gürtler V, Mayall BC (2001) Genomic approaches to typing, taxonomy and evolution of bacterial isolates. Int J Syst Evol Microbiol 51:3–16
Hammes WP, Vogel RF (1995) The genus Lactobacillus. In: Wood BJB, Holzapfel W (eds) The genera of lactic acid bacteria, vol 2. Chapman & Hall, Glasgow, pp 19–54
Hammes WP, Weiss N, Holzapfel W (1992) The genera Lactobacillus and Carnobacterium. In: Balows A, Trüpper HG, Dworkin M, Harder W, Schleifer KH (eds) The Prokaryotes: a handbook on the biology of bacteria: ecophysiology, isolation, identification, applications, vol 2. Springer-Verlag, New York, pp 1535–1594
Klein G (2007) International Committee on Systematics of Prokaryotes; Subcommittee on the taxonomy of Bifidobacterium, Lactobacillus and related organisms: Minutes of the meetings, 30 August and 1 September 2006, Bologna, Italy. Int J Syst Evol Microbiol 57:1367–1369
Marsh P, Martin VM (eds) (1999) The resident oral microflora. In: Oral microbiology, 4th edn. Wright, Oxford, pp 17–33
Massi M, Vitali B, Federici F, Matteuzzi D, Brigidi P (2004) Identification method based on PCR combined with automated ribotyping for tracking probiotic Lactobacillus strains colonizing the human gut and vagina. J Appl Microbiol 96:777–786
Olive DM, Bean P (1999) Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol 37:1661–1669
Rantsiou K, Drosinos EH, Gialitaki M, Metaxopoulos I, Comi G, Cocolin L (2006) Use of molecular tools to characterize Lactobacillus spp. isolated from Greek traditional fermented sausages. Int J Food Microbiol 112:215–222
Romesburg HC (ed) (1984) Overview of cluster analysis. In: Cluster analysis for researchers. Lifetime Learning Publications, Belmont, pp 7–73
Rossetti L, Giraffa G (2005) Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases. J Microbiol Methods 63:135–144
Ryu CS, Czajka JW, Sakamoto M, Benno Y (2001) Characterization of the Lactobacillus casei group and the Lactobacillus acidophilus group by automated ribotyping. Microbiol Immunol 45:271–275
Švec P, Sedláček I (2008) Characterization of Lactococcus lactis subsp. lactis isolated from surface waters. Folia Microbiol 53:53–56
Švec P, Dráb V, Sedláček I (2005) Ribotyping of Lactobacillus casei group strains isolated from dairy products. Folia Microbiol 50:223–228
Švec P, Sedláček I, Žáčková L, Nováková D, Kukletová M (2009) Lactobacillus spp. associated with early childhood caries. Folia Microbiol 54:53–58
Torriani S, Zapparoli G, Dellaglio F (1999) Use of PCR-based methods for rapid differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis. Appl Environ Microbiol 65:4351–4356
Tynkkynen S, Satokari R, Saarela M, Mattila-Sandholm T, Saxelin M (1999) Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains. Appl Environ Microbiol 65:3908–3914
van Houte J (1994) Role of micro-organisms in caries etiology. J Dent Res 73:672–681
Acknowledgments
Financial support from the Czech Science Foundation (project 310/09/0657) and the Ministry of Education, Youth and Sports of the Czech Republic (projects 1M0528 and MSM0021622416) is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Švec, P., Kukletová, M. & Sedláček, I. Comparative evaluation of automated ribotyping and RAPD-PCR for typing of Lactobacillus spp. occurring in dental caries. Antonie van Leeuwenhoek 98, 85–92 (2010). https://doi.org/10.1007/s10482-010-9432-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-010-9432-6