Journal of Periodontal and Implant Science

  • 제48권4호
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  • Pages.261-271
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  • 2018
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  • 2093-2278(pISSN)
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  • 2093-2286(eISSN)
대한치주과학회 (Korean Academy of Periodontology)
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Real-time PCR quantification of 9 periodontal pathogens in saliva samples from periodontally healthy Korean young adults

  • Choi, Heeyoung (Department of Periodontology, Institute of Translational Dental Sciences, Pusan National University School of Dentistry) ;
  • Kim, Eunhye (Gerotech Inc.) ;
  • Kang, Jihoon (Gerotech Inc.) ;
  • Kim, Hyun-Joo (Department of Periodontology, Institute of Translational Dental Sciences, Pusan National University School of Dentistry) ;
  • Lee, Ju-Youn (Department of Periodontology, Institute of Translational Dental Sciences, Pusan National University School of Dentistry) ;
  • Choi, Jeomil (Department of Periodontology, Institute of Translational Dental Sciences, Pusan National University School of Dentistry) ;
  • Joo, Ji-Young (Department of Periodontology, Institute of Translational Dental Sciences, Pusan National University School of Dentistry)
  • 투고 : 2018.07.24
  • 심사 : 2018.08.20
  • 발행 : 2018.08.30
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초록

Purpose: Few studies have examined periodontal pathogens from saliva samples in periodontally healthy young adults. The purposes of this study were to determine the prevalence of periodontopathic bacteria and to quantify periodontal pathogens in saliva samples using real-time polymerase chain reaction (PCR) assays in periodontally healthy Korean young adults under 35 years of age. Methods: Nine major periodontal pathogens were analyzed by real-time PCR in saliva from 94 periodontally healthy young adults. Quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Peptostreptococcus anaerobius, and Eikenella corrodens was performed by DNA copy number measurement. Results: F. nucleatum and E. corrodens were detected in all subjects; the numbers of positive samples were 87 (92.6%), 91 (96.8%), and 90 (95.7%) for P. gingivalis, P. anaerobius, and C. rectus, respectively. Other pathogens were also detected in periodontally healthy subjects. Analysis of DNA copy numbers revealed that the most abundant periodontal pathogen was F. nucleatum, which was significantly more prevalent than all other bacteria (P<0.001), followed by P. anaerobius, P. gingivalis, E. corrodens, C. rectus, and T. denticola. There was no significant difference in the prevalence of each bacterium between men and women. The DNA copy number of total bacteria was significantly higher in men than in women. Conclusions: Major periodontal pathogens were prevalent in the saliva of periodontally healthy Korean young adults. Therefore, we suggest that the development of periodontal disease should not be overlooked in periodontally healthy young people, as it can arise due to periodontal pathogen imbalance and host susceptibility.

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참고문헌

  1. Mark Bartold P, Van Dyke TE. Host modulation: controlling the inflammation to control the infection. Periodontol 2000 2017;75:317-29. https://doi.org/10.1111/prd.12169
  2. Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in subgingival plaque. J Clin Periodontol 1998;25:134-44. https://doi.org/10.1111/j.1600-051X.1998.tb02419.x
  3. Cullinan MP, Hamlet SM, Westerman B, Palmer JE, Faddy MJ, Seymour GJ. Acquisition and loss of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia over a 5-year period: effect of a triclosan/copolymer dentifrice. J Clin Periodontol 2003;30:532-41. https://doi.org/10.1034/j.1600-051X.2003.00292.x
  4. Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 2005;43:5721-32. https://doi.org/10.1128/JCM.43.11.5721-5732.2005
  5. Haffajee AD, Socransky SS. Microbial etiological agents of destructive periodontal diseases. Periodontol 2000 1994;5:78-111. https://doi.org/10.1111/j.1600-0757.1994.tb00020.x
  6. Socransky SS, Haffajee AD. Dental biofilms: difficult therapeutic targets. Periodontol 2000 2002;28:12-55. https://doi.org/10.1034/j.1600-0757.2002.280102.x
  7. Yang EY, Tanner AC, Milgrom P, Mokeem SA, Riedy CA, Spadafora AT, et al. Periodontal pathogen detection in gingiva/tooth and tongue flora samples from 18- to 48-month-old children and periodontal status of their mothers. Oral Microbiol Immunol 2002;17:55-9. https://doi.org/10.1046/j.0902-0055.2001.00092.x
  8. Tanner AC, Milgrom PM, Kent R Jr, Mokeem SA, Page RC, Riedy CA, et al. The microbiota of young children from tooth and tongue samples. J Dent Res 2002;81:53-7. https://doi.org/10.1177/002203450208100112
  9. Kononen E. Pigmented Prevotella species in the periodontally healthy oral cavity. FEMS Immunol Med Microbiol 1993;6:201-5. https://doi.org/10.1111/j.1574-695X.1993.tb00327.x
  10. McClellan DL, Griffen AL, Leys EJ. Age and prevalence of Porphyromonas gingivalis in children. J Clin Microbiol 1996.34:2017-9.
  11. Kamma JJ, Diamanti-Kipioti A, Nakou M, Mitsis FJ. Profile of subgingival microbiota in children with mixed dentition. Oral Microbiol Immunol 2000;15:103-11. https://doi.org/10.1034/j.1399-302x.2000.150206.x
  12. Lamell CW, Griffen AL, McClellan DL, Leys EJ. Acquisition and colonization stability of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in children. J Clin Microbiol 2000;38:1196-9.
  13. Loomer PM. Microbiological diagnostic testing in the treatment of periodontal diseases. Periodontol 2000 2004;34:49-56. https://doi.org/10.1046/j.0906-6713.2002.003424.x
  14. Sanz M, Lau L, Herrera D, Morillo JM, Silva A. Methods of detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in periodontal microbiology, with special emphasis on advanced molecular techniques: a review. J Clin Periodontol 2004;31:1034-47. https://doi.org/10.1111/j.1600-051X.2004.00609.x
  15. Gohler A, Hetzer A, Holtfreter B, Geisel MH, Schmidt CO, Steinmetz I, et al. Quantitative molecular detection of putative periodontal pathogens in clinically healthy and periodontally diseased subjects. PLoS One 2014;9:e99244. https://doi.org/10.1371/journal.pone.0099244
  16. Junemann S, Prior K, Szczepanowski R, Harks I, Ehmke B, Goesmann A, et al. Bacterial community shift in treated periodontitis patients revealed by ion torrent 16S rRNA gene amplicon sequencing. PLoS One 2012;7:e41606. https://doi.org/10.1371/journal.pone.0041606
  17. Ammann TW, Bostanci N, Belibasakis GN, Thurnheer T. Validation of a quantitative real-time PCR assay and comparison with fluorescence microscopy and selective agar plate counting for species-specific quantification of an in vitro subgingival biofilm model. J Periodontal Res 2013;48:517-26. https://doi.org/10.1111/jre.12034
  18. Shet UK, Oh HK, Kim HJ, Chung HJ, Kim YJ, Kim OS, et al. Quantitative analysis of periodontal pathogens present in the saliva of geriatric subjects. J Periodontal Implant Sci 2013;43:183-90. https://doi.org/10.5051/jpis.2013.43.4.183
  19. Umeda M, Contreras A, Chen C, Bakker I, Slots J. The utility of whole saliva to detect the oral presence of periodontopathic bacteria. J Periodontol 1998;69:828-33. https://doi.org/10.1902/jop.1998.69.7.828
  20. Kim JJ, Kim CJ, Camargo PM. Salivary biomarkers in the diagnosis of periodontal diseases. J Calif Dent Assoc 2013.41:119-24.
  21. Haffajee AD, Bogren A, Hasturk H, Feres M, Lopez NJ, Socransky SS. Subgingival microbiota of chronic periodontitis subjects from different geographic locations. J Clin Periodontol 2004;31:996-1002. https://doi.org/10.1111/j.1600-051X.2004.00597.x
  22. Darveau RP, Hajishengallis G, Curtis MA. Porphyromonas gingivalis as a potential community activist for disease. J Dent Res 2012;91:816-20. https://doi.org/10.1177/0022034512453589
  23. Feres M, Teles F, Teles R, Figueiredo LC, Faveri M. The subgingival periodontal microbiota of the aging mouth. Periodontol 2000 2016;72:30-53. https://doi.org/10.1111/prd.12136
  24. Zhuang LF, Watt RM, Mattheos N, Si MS, Lai HC, Lang NP. Periodontal and peri-implant microbiota in patients with healthy and inflamed periodontal and peri-implant tissues. Clin Oral Implants Res 2016;27:13-21.
  25. Testa M, Ruiz de Valladares R, Benito de Cardenas IL. Correlation between bacterial counts in saliva and subgingival plaque. Acta Odontol Latinoam 1999;12:63-74.
  26. Sakamoto M, Takeuchi Y, Umeda M, Ishikawa I, Benno Y. Rapid detection and quantification of five periodontopathic bacteria by real-time PCR. Microbiol Immunol 2001;45:39-44. https://doi.org/10.1111/j.1348-0421.2001.tb01272.x
  27. Marsh PD, Moter A, Devine DA. Dental plaque biofilms: communities, conflict and control. Periodontol 2000 2011;55:16-35. https://doi.org/10.1111/j.1600-0757.2009.00339.x
  28. Arenas Rodrigues VA, de Avila ED, Nakano V, Avila-Campos MJ. Qualitative, quantitative and genotypic evaluation of Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum isolated from individuals with different periodontal clinical conditions. Anaerobe 2018;52:50-8. https://doi.org/10.1016/j.anaerobe.2018.05.015
  29. Zhou X, Liu X, Li J, Aprecio RM, Zhang W, Li Y. Real-time PCR quantification of six periodontal pathogens in saliva samples from healthy young adults. Clin Oral Investig 2015;19:937-46. https://doi.org/10.1007/s00784-014-1316-0
  30. Papapanou PN, Neiderud AM, Papadimitriou A, Sandros J, Dahlen G. "Checkerboard" assessments of periodontal microbiota and serum antibody responses: a case-control study. J Periodontol 2000;71:885-97. https://doi.org/10.1902/jop.2000.71.6.885
  31. Griffen AL, Becker MR, Lyons SR, Moeschberger ML, Leys EJ. Prevalence of Porphyromonas gingivalis and periodontal health status. J Clin Microbiol 1998;36:3239-42.
  32. Amano A, Kuboniwa M, Nakagawa I, Akiyama S, Morisaki I, Hamada S. Prevalence of specific genotypes of Porphyromonas gingivalis fimA and periodontal health status. J Dent Res 2000;79:1664-8. https://doi.org/10.1177/00220345000790090501
  33. Hyvarinen K, Laitinen S, Paju S, Hakala A, Suominen-Taipale L, Skurnik M, et al. Detection and quantification of five major periodontal pathogens by single copy gene-based real-time PCR. Innate Immun 2009;15:195-204. https://doi.org/10.1177/1753425908101920
  34. Haffajee AD, Japlit M, Bogren A, Kent RL Jr, Goodson JM, Socransky SS. Differences in the subgingival microbiota of Swedish and USA subjects who were periodontally healthy or exhibited minimal periodontal disease. J Clin Periodontol 2005;32:33-9. https://doi.org/10.1111/j.1600-051X.2004.00624.x
  35. Lira-Junior R, Akerman S, Klinge B, Bostrom EA, Gustafsson A. Salivary microbial profiles in relation to age, periodontal, and systemic diseases. PLoS One 2018;13:e0189374. https://doi.org/10.1371/journal.pone.0189374
  36. Nonnenmacher C, Dalpke A, Rochon J, Flores-de-Jacoby L, Mutters R, Heeg K. Real-time polymerase chain reaction for detection and quantification of bacteria in periodontal patients. J Periodontol 2005;76:1542-9. https://doi.org/10.1902/jop.2005.76.9.1542
  37. Papaioannou W, Gizani S, Haffajee AD, Quirynen M, Mamai-Homata E, Papagiannoulis L. The microbiota on different oral surfaces in healthy children. Oral Microbiol Immunol 2009;24:183-9. https://doi.org/10.1111/j.1399-302X.2008.00493.x
  38. Bartold PM, Van Dyke TE. Periodontitis: a host-mediated disruption of microbial homeostasis. Unlearning learned concepts. Periodontol 2000 2013;62:203-17. https://doi.org/10.1111/j.1600-0757.2012.00450.x
  39. Preshaw PM, Taylor JJ. How has research into cytokine interactions and their role in driving immune responses impacted our understanding of periodontitis? J Clin Periodontol 2011;38 Suppl 11:60-84. https://doi.org/10.1111/j.1600-051X.2010.01671.x
  40. Ebersole JL, Dawson DR 3rd, Morford LA, Peyyala R, Miller CS, Gonzalez OA. Periodontal disease immunology: 'double indemnity' in protecting the host. Periodontol 2000 2013;62:163-202. https://doi.org/10.1111/prd.12005

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