Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
권호 표지
DOI QR코드

DOI QR Code

IDENTIFICATION OF PUTATIVE PATHOGENS IN ACUTE ENDODONTIC INFECTIONS BY PCR BASED ON 16S rDNA

중합효소연쇄반응법을 이용한 급성 치수 및 치근단 질환의 병원성 세균의 동정

  • Kim, Ji-Hoon (Department of Conservative Dentistry, College of Dentistry, Chosun University) ;
  • Yoo, So-Young (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Lim, Sun-A (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Kook, Joong-Ki (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Lim, Sang-Soo (Department of Conservative Dentistry, College of Dentistry, Chosun University) ;
  • Park, Seul-Hee (Department of Conservative Dentistry, College of Dentistry, Chosun University) ;
  • Hwang, Ho-Keel (Department of Conservative Dentistry, College of Dentistry, Chosun University)
  • 김지훈 (조선대학교 치과대학 보존학교실) ;
  • 유소영 (조선대학교 치과대학 구강생화학교실, 구강생물학연구소) ;
  • 임선아 (조선대학교 치과대학 구강생화학교실) ;
  • 국중기 (조선대학교 치과대학 구강생화학교실, 구강생물학연구소) ;
  • 임상수 (조선대학교 치과대학 보존학교실) ;
  • 박슬희 (조선대학교 치과대학 보존학교실) ;
  • 황호길 (조선대학교 치과대학 보존학교실, 구강생물학연구소)
  • Published : 2003.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

The purpose of this study was to investigate the frequency of 7 putative pathogens in endodontic infections. The specimens were collected from infected pulpal tissue of patients who were referred for root canal treatment to the department of conservative dentistry, Chosun University Samples were collected aseptically using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ml of 1 X PBS. DNAs were extracted from the samples by direct DNA extraction method using lysis buffer (0.5% EDTA, 1% Triton X-100). Identification of 7 putative pathogens was performed by PCR based on 16S rDNA. The target species were as follows : Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, and Treponema denticola. Our data revealed that the prevalence of P. endodontalis was found in 88.6% (39/54), P. ginivalis 52.3% (23/44), P. nigrescens 18.2% (8/44), P intermedia 15.9% (7/44) B. forsythus 18.2% (8/44), A. actinomycetemcomitans 3.3% (1/44), T. denticola 25% (l1/44) of the samples. The high prevalence of P. endodontalis and P. ginivalis suggests that they may play an important role in the etiology of endodontic infections.

Keywords

References

  1. Miller WD. Microorganisms of the human mouse. Philadelphia: S.S. White Dental Co., 1890
  2. Kakehashi S. Stanley HR and Fitzgerald RJ. The effects of surgical exposures of dental pulps in germ-free and conventional laboratory rats. Oral. Surg. Oral. Med. Oral. Pathol. 20:340-349, 1965 https://doi.org/10.1016/0030-4220(65)90166-0
  3. Sundqvist G. Bacteriological studies of necrotic dental pulps. Umea: Umea University. 1976
  4. Brown TA. Molecular biology labfax Ⅱ. San Diego: Academic press, 1998
  5. Ashimoto A. Chen C. Bakker I and Slots J. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesion. Oral. Microbiol, Immunol 11: 56-61. 1996
  6. Brook I. Frazier EH and Gher ME. Aerobic and anaerobic microbiology of periapical abscess. Oral. Microbiol. lmmunol 6: 123-125, 1991 https://doi.org/10.1111/j.1399-302X.1991.tb00464.x
  7. Socransky SS and Haffajee AD, Clinical periodontology and implant dentstry, 3rd eds. Copenhagen: Munksgaard, 138-188. 1997
  8. Sundqvist GK. Eckerbom MI. Larsson AP and Sjogren UF, Capacity of anaerobic bacteria from necrotic dental pulps to induce purulent infections. Infect Immun 25: 685-693, 1979
  9. Van Winkelhoff AJ. Carlee AW and De Graaff J. Bacteriodes endodontalis and others black-pigmented Bacteriodes species in odontogenic abscesses. Infect Immun 49: 494-498. 1985
  10. Haapasalo M. Ranta H. Ranta K and Shah H. Black-pigmented bacteroides spp. in human apical periodontitis. Infect Immun 53: 149-153, 1986
  11. Siqueira JF. Rocas IN. Oliveira JCM and Santos -RN. Molecular detection of black-pigmented bactena Ⅲ infections of endodontic origin. J Endod 27: 563-566, 2001 https://doi.org/10.1097/00004770-200109000-00002
  12. Sundqvist G. Johansson E and Rocas IN. Prevalence of black-pigmented bacteria with endodontic infections. J Endod 15:13-19, 1989 https://doi.org/10.1016/S0099-2399(89)80092-5
  13. Van Winkelhoff AJ. The role of black-pigmented bacteroides in human oral infections. J Clin Periodontol 15: 145-155, 1988 https://doi.org/10.1111/j.1600-051X.1988.tb01561.x
  14. Julio Cezar Machado de Oliveira. Jose Freitas Siqueira Jr. Gabriela B. Alves Raphael Hirata Jr. Arnaldo FB and Andrade. Detection of Porphyromonas endodontalis in infected root canals by 16S rRNA gene- directed Polymerase chain reaction. J Endod 26:729-732. 2000 https://doi.org/10.1097/00004770-200012000-00016
  15. Wahlfors J and Meurman JH. Simultaneous detection of Actinobacillus actinomycetemcomitans and Porphylomonas gingivalis by a rapid PCR method. J Dent Res 74: 1796-1801. 1995 https://doi.org/10.1177/00220345950740111301
  16. Baumgartner JC. Watkins BJ, Bae KS and Xia T. Association of black-pigmented bacteria with endodontic infections. J Endod 25: 413-415. 1999 https://doi.org/10.1016/S0099-2399(99)80268-4
  17. Haapasalo M. Bacteroides spp. in dental root canal infections. Endod Dent Treumsiol S: 1-10. 1989
  18. Siqueira JF. Rocas IN. Oliveira JCM and Santos KRN. Detection of putative oral pathogens in acute periradicular abscesses by 16S rDNA-directed Polymerase chain reaction. J Endod 27:164-167, 2001 https://doi.org/10.1097/00004770-200103000-00004
  19. Hashioka K. Yamasaki M. Nakane A. Horiba. N and Nakamura H. The relationship between clinical symptoms and anaerobic bacteria from infected root canals. J Endod 18: 558-561. 1971
  20. Shah HN and Gharbia SE. Biochemical and chemical studies on strains designated Prevotella intermedia and proposal of a new pigmented species. Prevotella nigrescens sp. nov. Int J Syst Bacteriol 42: 542-546. 1992 https://doi.org/10.1099/00207713-42-4-542
  21. Gharbia SE. Haapasalo M and Shah HN. Characterization of Prevotella intermedia and Prevotella nigrescens isolates from periodontic and endodontic infection. J Periodontol Sb: 56-61. 1994
  22. Conrads G. Mutters R. Fischer J, Brauner A. Lutticken R and Lampert F. PCR reaction and dot-blot hybridization to monitor the distribution of oral pathogens within plaque samples of periodontally healthy individuals. J Periodontol 67: 995-1003. 1996
  23. Dahlen G, Wikstrom M. Renvert S, Gmur R. and Guggenheim B. Biochemical and serological characterization of Bacteroides intermedius strains isolated from the deep periodontal pocket. J Clin Microbiol 28: 2269-2274. 1990
  24. Matto J, Asikainen Sand Vaisanen ML. Role of Porphyromonas gingivalis. Prevotella intermedia, and Prevotella nigrescens in extraoral and some odontogenic infections. Clin Infect Dis 25 Suppl S194-S198. 1997 https://doi.org/10.1086/516205
  25. Milson SE, Sprague SV. Dymock D. Weighman AJ and Wade WG. Rapid differentiation of Prevotella intermedia and Prevotella nigrescens by 16S rDNA PCR-RFLP. J Med Microbiol. 44: 41-43. 1992
  26. Dahle UR, Tronstad L and Olsen I. Observation of an unusually large spirochete in endodontic infection. Oral Microbiol and Immunol 8: 251-253. 1993 https://doi.org/10.1111/j.1399-302X.1993.tb00569.x
  27. Hampp. EG. Isolation and identification of spirochetes obtained from unexposed canals of pulp-involved teeth. Oral Surg Oral Med and Oral Pathol 10: 1100-1104. 1957 https://doi.org/10.1016/0030-4220(57)90062-2
  28. Thilo BE. Bachni P and Holz J. Darkfield observation of bacterial distribution in root canals following pulp necrosis. J Endod 12: 202-205, 1986 https://doi.org/10.1016/S0099-2399(86)80155-8
  29. Trope M, Tronstad L. Rosenberg ES and Litsgarten MA. Darkfield microscopy as a diagnostic aid in differentiating exudates from endodontic and periodontal abscesses. J Endod 14: 35-39, 1984 https://doi.org/10.1016/S0099-2399(88)80239-5
  30. Siqueira JF. Rocas IN. Moraes SR and Santos KRN. Direct amplification of rRNA gene sequences for identification of selected oral pathogens in root canal infections. Int Endod J 35: 345-351. 2002 https://doi.org/10.1046/j.1365-2591.2002.00485.x
  31. Rocas IN. Siqueira JF, Santos KRN and Coelho M.A. 'Red complex' (Bacteroides forsyth us. Porphylomonas gingivalis. and Treponema denticole) in endodontic infections: A molecular approach. Oral. Surg. Oral. Med. Oral. Pathol. Oral Radiol Endod 91: 468-471. 2001 https://doi.org/10.1067/moe.2001.114379
  32. Fenno JC and McBride BC. Virulence factor of oral treponemes. Anaerobe 4: 1-17. 1998 https://doi.org/10.1006/anae.1997.0131
  33. Gon alves RB and Mouton C. Molecular detection of bacteriodes forsythus in infected root canals. J Endod 25: 336-340. 1999 https://doi.org/10.1016/S0099-2399(06)81167-2
  34. Conrads G. Gharbia SF. Gulabivala K. Lampert F and Shah HN. The use of a 16S rDNA PCR for the detection of endodontopathogenic bacteria. J Endod 23: 433-438. 1997 https://doi.org/10.1016/S0099-2399(97)80297-X
  35. Siqueira JF. Rocas IN, Souto R, De Uzeda M and Colombo AP. Microbiological evaluation of acute periradicular abscesses by DNA-DNA hybridization. Oral Surg Oral Med. Oral Pathol, Oral Radiol Endod 92: 451-457, 2001 https://doi.org/10.1067/moe.2001.118620
  36. Takemoto T, Kurihara H and Dahlen G. Characterization of Bacteriodes forsythus isolates. J Clin Microbiol 35: 1378-1381. 1997
  37. Meyer DH and Fives-Taylor PM. The role of Actinobacillus actinomycetemcomitans in the pathogenesis of periodontal disease. Tren In Microbiol 5: 224-228. 1997 https://doi.org/10.1016/S0966-842X(97)01055-X
  38. Siqueira JF, Rocas IN, Souto R. Uzeda M and Colombo AP. Checkerboard DNA-DNA hybridization analysis of endodontic infections. Oral Surg. Oral Med. Oral Pathol. Oral. Radiol. Endod 89: 744-748. 2000 https://doi.org/10.1067/moe.2000.106576
  39. Sunde PT, Tronstad L. Eribe ER. Lind PO and Olsen I. Assessment of periradicular microbiota by DNA-DNA hybridization. Endod Dent Traumatol 16: 191-196, 2000 https://doi.org/10.1034/j.1600-9657.2000.016005191.x