Jundishapur Journal of Microbiology

Published by: Kowsar

Development and Comparison of Conventional PCR and SYBR Green Real Time PCR for Detection of Aggregatibacteractinomycetemcomitans and Tannerella forsythensis

Mohammad Soleimani 1 , * , Mohammad Reza Zolfaghari 1 and Abbas Morovvati 1
Authors Information
1 Department of Microbiology, Qom Branch, Islamic Azad University, Qom, IR Iran
Article information
  • Jundishapur Journal of Microbiology: October 01, 2013, 6 (8); e6757
  • Published Online: October 1, 2013
  • Article Type: Research Article
  • Received: June 13, 2012
  • Revised: August 25, 2012
  • Accepted: October 2, 2012
  • DOI: 10.5812/jjm.6757

To Cite: Soleimani M, Zolfaghari M R, Morovvati A. Development and Comparison of Conventional PCR and SYBR Green Real Time PCR for Detection of Aggregatibacteractinomycetemcomitans and Tannerella forsythensis, Jundishapur J Microbiol. 2013 ; 6(8):e6757. doi: 10.5812/jjm.6757.

Copyright © 2013, Ahvaz Jundishapur University of Medical Sciences. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
1. Background
2. Objectives
3. Materials and Methods
4. Results
5. Discussion
  • 1. Psoter WJ, Ge Y, Russell SL, Chen Z, Katz RV, Jean-Charles G, et al. PCR detection of Streptococcus mutans and Aggregatibacter actinomycetemcomitans in dental plaque samples from Haitian adolescents. Clin Oral Investig. 2011; 15(4): 461-9[DOI][PubMed]
  • 2. Paster BJ, Boches SK, Galvin JL, Ericson RE, Lau CN, Levanos VA, et al. Bacterial diversity in human subgingival plaque. J Bacteriol. 2001; 183(12): 3770-83[DOI][PubMed]
  • 3. Socransky SS, Haffajee AD, Ximenez-Fyvie LA, Feres M, Mager D. Ecological considerations in the treatment of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis periodontal infections. Periodontol 2000. 1999; 20: 341-62[PubMed]
  • 4. Kumar PS, Griffen AL, Moeschberger ML, Leys EJ. Identification of candidate periodontal pathogens and beneficial species by quantitative 16S clonal analysis. J Clin Microbiol. 2005; 43(8): 3944-55[DOI][PubMed]
  • 5. Slots J, Ting M. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in human periodontal disease: occurrence and treatment. Periodontol 2000. 1999; 20: 82-121[PubMed]
  • 6. Kozarov EV, Dorn BR, Shelburne CE, Dunn WA, Jr, Progulske-Fox A. Human atherosclerotic plaque contains viable invasive Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Arterioscler Thromb Vasc Biol. 2005; 25(3)-8[DOI][PubMed]
  • 7. Nonnenmacher C, Dalpke A, Mutters R, Heeg K. Quantitative detection of periodontopathogens by real-time PCR. J Microbiol Methods. 2004; 59(1): 117-25[DOI][PubMed]
  • 8. Shelburne CE, Prabhu A, Gleason RM, Mullally BH, Coulter WA. Quantitation of Bacteroides forsythus in subgingival plaque comparison of immunoassay and quantitative polymerase chain reaction. J Microbiol Methods. 2000; 39(2): 97-107[PubMed]
  • 9. Simonson LG, McMahon KT, Childers DW, Morton HE. Bacterial synergy of Treponema denticola and Porphyromonas gingivalis in a multinational population. Oral Microbiol Immunol. 1992; 7(2): 111-2[PubMed]
  • 10. Socransky SS, Haffajee AD, Dzink JL, Hillman JD. Associations between microbial species in subgingival plaque samples. Oral Microbiol Immunol. 1988; 3(1): 1-7[PubMed]
  • 11. Hashimoto M, Ogawa S, Asai Y, Takai Y, Ogawa T. Binding of Porphyromonas gingivalis fimbriae to Treponema denticola dentilisin. FEMS Microbiol Lett. 2003; 226(2): 267-71[PubMed]
  • 12. Suzuki N, Yoshida A, Saito T, Kawada M, Nakano Y. Quantitative microbiological study of subgingival plaque by real-time PCR shows correlation between levels of Tannerella forsythensis and Fusobacterium spp. J Clin Microbiol. 2004; 42(5): 2255-7[PubMed]
  • 13. Slots J. Rapid identification of important periodontal microorganisms by cultivation. Oral Microbiol Immunol. 1986; 1(1): 48-57[PubMed]
  • 14. Bonta Y, Zambon JJ, Genco RJ, Neiders ME. Rapid identification of periodontal pathogens in subgingival plaque: comparison of indirect immunofluorescence microscopy with bacterial culture for detection of Actinobacillus actinomycetemcomitans. J Dent Res. 1985; 64(5): 793-8[PubMed]
  • 15. Savitt ED, Strzempko MN, Vaccaro KK, Peros WJ, French CK. Comparison of cultural methods and DNA probe analyses for the detection of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius in subgingival plaque samples. J Periodontol. 1988; 59(7): 431-8[DOI][PubMed]
  • 16. Alsina M, Olle E, Frias J. Improved, low-cost selective culture medium for Actinobacillus actinomycetemcomitans. J Clin Microbiol. 2001; 39(2): 509-13[DOI][PubMed]
  • 17. Ramseier CA, Kinney JS, Herr AE, Braun T, Sugai JV, Shelburne CA, et al. Identification of pathogen and host-response markers correlated with periodontal disease. J Periodontol. 2009; 80(3): 436-46[DOI][PubMed]
  • 18. Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CM, Savelkoul PH. The additional value of real-time PCR in the quantitative detection of periodontal pathogens. J Clin Periodontol. 2006; 33(6): 427-33[DOI][PubMed]
  • 19. Yoshida A, Suzuki N, Nakano Y, Oho T, Kawada M, Koga T. Development of a 5' fluorogenic nuclease-based real-time PCR assay for quantitative detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. J Clin Microbiol. 2003; 41(2): 863-6[PubMed]
  • 20. Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CM, Savelkoul PH. Comparison of real-time PCR and culture for detection of Porphyromonas gingivalis in subgingival plaque samples. J Clin Microbiol. 2003; 41(11): 4950-4[PubMed]
  • 21. 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(9): 1664-8[PubMed]
  • 22. Ashimoto A, Chen C, Bakker I, Slots J. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions. Oral Microbiol Immunol. 1996; 11(4): 266-73[PubMed]
  • 23. Oho T, Yamashita Y, Shimazaki Y, Kushiyama M, Koga T. Simple and rapid detection of Streptococcus mutans and Streptococcus sobrinus in human saliva by polymerase chain reaction. Oral Microbiol Immunol. 2000; 15(4): 258-62[PubMed]
  • 24. Suzuki N, Nakano Y, Yoshida Y, Ikeda D, Koga T. Identification of Actinobacillus actinomycetemcomitans serotypes by multiplex PCR. J Clin Microbiol. 2001; 39(5): 2002-5[DOI][PubMed]
  • 25. Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CM, Savelkoul PH. Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR. FEMS Immunol Med Microbiol. 2005; 45(2): 191-9[DOI][PubMed]
  • 26. Orru G, Marini MF, Ciusa ML, Isola D, Cotti M, Baldoni M, et al. Usefulness of real time PCR for the differentiation and quantification of 652 and JP2 Actinobacillus actinomycetemcomitans genotypes in dental plaque and saliva. BMC Infect Dis. 2006; 6: 98[DOI][PubMed]
  • 27. Sambrook J, Russell DW. Molecular Cloning: A Laboratory Manual. 2001;
  • 28. Chiang YC, Yang CY, Li C, Ho YC, Lin CK, Tsen HY. Identification of Bacillus spp., Escherichia coli, Salmonella spp., Staphylococcus spp. and Vibrio spp. with 16S ribosomal DNA-based oligonucleotide array hybridization. Int J Food Microbiol. 2006; 107(2): 131-7[DOI][PubMed]
  • 29. Dumitrescu AL. Etiology and Pathogenesis of Periodontal Disease. 2010;
  • 30. Soder B, Yakob M, Meurman JH, Andersson LC, Klinge B, Soder PO. Periodontal disease may associate with breast cancer. Breast Cancer Res Treat. 2011; 127(2): 497-502[DOI][PubMed]
  • 31. Tonjum T, Haas R. Identification of Actinobacillus actinomycetemcomitans by leukotoxin gene-specific hybridization and polymerase chain reaction assays. J Clin Microbiol. 1993; 31(7): 1856-9[PubMed]
  • 32. Leblebicioglu B, Kulekci G, Ciftci S, Keskin F, Badur S. Salivary detection of periodontopathic bacteria and periodontal health status in dental students. Anaerobe. 2009; 15(3): 82-6[DOI][PubMed]
  • 33. D'Ercole S, Catamo G, Tripodi D, Piccolomini R. Comparison of culture methods and multiplex PCR for the detection of periodontopathogenic bacteria in biofilm associated with severe forms of periodontitis. New Microbiol. 2008; 31(3): 383-91[PubMed]
  • 34. Flemmig TF, Rudiger S, Hofmann U, Schmidt H, Plaschke B, Stratz A, et al. Identification of Actinobacillus actinomycetemcomitans in subgingival plaque by PCR. J Clin Microbiol. 1995; 33(12): 3102-5[PubMed]
  • 35. Conrads G, Flemmig TF, Seyfarth I, Lampert F, Lutticken R. Simultaneous detection of Bacteroides forsythus and Prevotella intermedia by 16S rRNA gene-directed multiplex PCR. J Clin Microbiol. 1999; 37(5): 1621-4[PubMed]
  • 36. Soleimani M, Eini F, Fallah Raufi M, Azari F, Farzampour S, Jamshidian E, et al. Design of Multiplex Polymerase Chain Reaction(PCR) Method for Molecular Detection of Yersinia pestis Bacterium. Yakhteh. 2010; 12(3): 363-370
  • 37. van Steenbergen TJ, Timmerman MF, Mikx FH, de Quincey G, van der Weijden GA, van der Velden U, et al. Discrepancy between culture and DNA probe analysis for the detection of periodontal bacteria. J Clin Periodontol. 1996; 23(10): 955-9[PubMed]
  • 38. Slots J, Chen C. Detection of Porphyromonas gingivalis associated with human periodontitis by DNA methods. Clin Infect Dis. 1993; 16 Suppl 4-8[PubMed]
  • 39. Riggio MP, Macfarlane TW, Mackenzie D, Lennon A, Smith AJ, Kinane D. Comparison of polymerase chain reaction and culture methods for detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in subgingival plaque samples. J Periodontal Res. 1996; 31(7): 496-501[PubMed]
Readers' Comments