Bacterial diversity in children's dental caries

소아의 치아 우식 부위별 세균 다양성

  • Kim, Eun-Mi (Department of Dental Hygiene, Gwangju Health University) ;
  • Baik, Keun-Sik (Division of Life Science, Korea Basic Science Institute) ;
  • Ha, Myung-Ok (Department of Dental Hygiene, Gwangju Health University)
  • 김은미 (광주보건대학교 치위생과) ;
  • 백근식 (한국기초과학지원연구원 생명과학연구부) ;
  • 하명옥 (광주보건대학교 치위생과)
  • Received : 2013.07.31
  • Accepted : 2013.10.17
  • Published : 2013.10.31


Objectives : Molecular biology techniques were employed to assess diversity of bacterial in children's dental caries. Methods : DNA of germs was extracted and the diversity of the 16S rRNA clones was analyzed by amplified rDNA restriction analysis and sequencing. The experimental samples were pit and fissure caries (PC), deep dentinal caries (DC), smooth surface caries (SC), and supragingival plaque (PQ) from 50 children of age less than 12 years old. The control group was healthy teeth supragingival plaque (HT). Thirty clones from each 16S rRNA clone library of 5 samples were randomly selected, thus a total of 150 clones were analyzed. Results : Amplified rDNA restriction analysis uncovered 18, 20, 11, 17, and 22 phylotypes from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and supragingival plaque, respectively. Sequencing analysis found the dominance of Actinomycs naeslundii and Fusobacterium nucleatum in the healthy teeth; Leptotrichia sp. in the pit and fissure caries; Actinomyces sp., Streptococcus mutans, and Rahnella aquatilis in the deep dentinal caries; Streptococcus mutans and Actinomyces sp. in the smooth surface caries; Enterobacter hormaechei and Streptococcus sanguinis in the supragingival plaque. Conclusions : Clonal analysis identified 6 phyla, 20 genera, and 51 species.


ARDRA;bacterial diversity;dental caires;16S rRNA gene


Supported by : 광주보건대학교


  1. Kang HS, Jeong JH. A study on the correlation between the dental caries and dietary habits and snack intake of pre-school children. J Korean Acad Dent Hyg 2010; 10: 345-59.
  2. Kim YG, Han MD. Oral microbiology. Seoul: Komoonsa; 1998: 261-84.
  3. Kook JK, Park JW, Yoo SY, Kim HS, LEE NY. Distribution of mutans streptococoi in dental plaque of children. J Korean Acad Pediatr Dent 2004; 31: 439-47.
  4. Woese CR. Bacterial evolution. Microbiol Rev 1987; 51: 221-71.
  5. Vaneechoutte M, Rossau R, De vos P, Gillis M, Janssens D, Paepe N, et al. Rapid identification of bacteria of the Camamonadaceae with amplified ribosomal DNA-restriction analysis(ARDRA). FEMS Microbial Lett 1992; 93: 227-33. DOI:10.1016/0378-1097(92)90467-3.
  6. Pitcher DG, Saunders NA, Owen RJ. Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett in App Microbiol 1989; 8: 151-6.
  7. Heyndrickx M, Vauterin L, Vandamme P, Kersters K, De Vos P. Applicability of combined amplified 16s rDNA restriction analysis(ARDRA) patterns in bacterial phylogeny and taxonomy. J Microbiol Methods 1996; 26: 247-59.
  8. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 1997; 25: 4876-82. DOI: 10.1093/nar/25.24.4876.
  9. Munro HN. Mammalian protein metabolism. New York: Academic Press; 1970: 21-132.
  10. Hugenholtz P, Tyson GW, Webb RI, Wagner AM, Blackall LL. Investigation of candidate division TM7, a recently recognized major lineage of the domain Bacteria with no known pure-culture representatives. Appl Environ Microbiol 2001; 67: 411-9. DOI: 10.1128/AEM.67.1.411-419.2001.
  11. Marcy Y, Ouverney C, Bik EM, Losekann T, Ivanova N, Martin HG, et al. Dissecting biological "dark matter" with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth. Proc Natl Acad Sci 2007; 104: 11889-94. DOI: 10.1073/pnas.0704662104.
  12. Curtis MA, Zenobia C, Darveau RP. The relationship of the oral microbiotia to periodontal health and disease. Cell Host Microbe 2011; 10: 302-6. DOI: 10.1016/j.chom.2011.09.008.
  13. Wade WG. The oral microbiome in health disease. Pharmacol Res 2013; 69: 137-43. DOI: 10.1016/j.phrs.2012.11.006.
  14. Liu B, Faller LL, Klitgord N, Mazumdar V, Ghodsi M, Sommer DD, et al. Deep sequencing of the oral microbiome reveals signatures of periodontal disease. PloS One 2012; e37919. DOI: 10.1371/journal.pone.0037919.
  15. Keijser BJ, Zaura E, Huse SM, van der Vossen JM, Schuren FH, Montijn RC, et al. Pyrosequencing analysis of the oral microflora of healthy adults. J Dent Res 2008; 87: 1016-20. DOI: 10.1177/154405910808701104.
  16. Li J, Helmerhorst EJ, Leone CW, Troxler RF, Yaskell T, Haffajee AD, et al. Identification of early microbial colonizers in human dental biofilm. J Appl Microbiol 2004; 97: 1311-8. DOI: 10.1111/j.1365-2672.2004.02420.x.
  17. Bolstad AI, Jensen HB, Bakken V. Taxonomy, biology, and periodontal aspects of Fusobacterium nucleatum. Cln Microbiol Rev 1996; 9: 55-71.
  18. Signat B, Roques C, Poulet P, Duffaut D. Fusobacterium nucleatum in periodontal health and disease. Curr Issues Mol Biol 2011; 13: 25-36.
  19. McKinnon A, Black AY, Lortie K, Fleming NA. A case of adolescent pelvic inflammatory disease caused by a rare bacterium: Fusobacterium. J Pediatr Adolesc Gynecol 2013; in press. DOI: 10.1016/j.jpag.2013.02.008.
  20. Cisar JL, Sandberg Al, Mergenhagen SE. The function and distribution of different fimbriae on strains of Acitnomyces viscosus and Actinomyces naeslundii. J Dent Res 1984; 63: 393-6. DOI: 10.1177/00220345840630030701.
  21. Eribe ERK, Paster BJ, Caugant DA, Dewhirst F E, Stromberg VK, Lacy GH, et al. Genetic diversity of Leptotrichia and description of Leptotrichia goodfellowii sp. nov., Leptotrichia hofstadii sp. nov., Leptotrichia shahii sp. nov. and Leptotrichia wadei sp. nov. Int J Syst Evol Microbiol 2004; 54: 583-92. DOI: 10.1099/ijs.0.02819-0.
  22. Bowden GHW. Microbilogy of root surface caries in humans. J Dent Res 1990; 69: 1205-10. DOI: 10.1177/00220345900690051701.
  23. Van Houte J, Jordan HV, Laraway R, Kent R, Soparkar PM, DePaola PF. Association of the microbial flora of dental plaque and saliva with human root-surface caries. J Dent Res 1990; 69: 1463-8. DOI: 10.1177/00220345900690080301.
  24. Chang CL, Jeong J, Shin JH, Lee EY, Son HC. Rahnella aquatilis sepsis in an immunocompetent adult. J Clin Microbiol 1999; 37: 4161-2.
  25. Tash K. Rahnella aquatilis bacteremia from a suspected urinary source. J Clin Microbiol 2005; 43: 2526-8.
  26. Leadbetter ER, Holt SC, Socransky SS. Capnocytophaga: new genus of gram-negative gliding bacteria. I. General characteristics, taxonomic considerations and significance. Arch Microbiol 1979; 122: 9-16.
  27. O'Hara CM, Steigerwalt AG, Hill BC, Farmer III JJ, Fanning GR, Brenner DJ. Enterobacter hormaechei, a new species of the family Enterobacteriaceae formerly known as Enteric Group 75. J Clin Microbiol 1989; 27: 2046-9.
  28. Hoffmann H, Stindl S, Ludwig W, Stumpf A, Mehlen A, Monget D, et al. Enterobacter hormaechei subsp. oharae subsp. nov., E. hormaechei subsp. hormaechei comb. nov., and E. hormaechei subsp. steigerwaltii subsp. nov., three new subspecies of clinical importance. J Clin Microbiol 2005; 43: 3297-303.