Banner Portal
Comparison of two different methods for detecting periodontal pathogenic bacteria
PDF (Português (Brasil))

Keywords

Periodontal diseases. Polymerase chain reaction. Bacteria.

How to Cite

1.
Bedran TBL, Oliveira GJPL de, Spolidorio LC, Cirelli JA, Spolidorio DP. Comparison of two different methods for detecting periodontal pathogenic bacteria. Braz. J. Oral Sci. [Internet]. 2017 Aug. 11 [cited 2024 May 24];15(3):166-72. Available from: https://periodicos.sbu.unicamp.br/ojs/index.php/bjos/article/view/8649599

Abstract

Aim: To perform a comparative analysis between two methods for detecting Porphyromonas gingivalis, Tannerella forsythia and Porphyromonas endodontalis in periodontal plaque samples. Methods: The study sample consisted of twenty systemically healthy patients showing generalized chronic periodontitis. The subgingival samples for microbiological analysis were collected before (baseline) and 60 days after a basic periodontal therapy from 30 non-adjacent affected sites (Probing Depth (PD): 5-7 mm, Clinical Attachment Loss (CAL) ≥ 5 mm, positive for Bleeding on Probing (BOP)). Microbiological analysis was performed by PCR and qPCR. To allow a comparative analysis between both methods, qPCR was divided in three different scores (score 2: presence of more than 100 bacteria; score 1: presence of 10-100 bacteria, and score 0: absence of bacteria), in accordance to DNA quantity, while for PCR two scores were assigned: presence or absence of bacteria. Results: qPCR demonstrated higher sensitivity in the detection of these pathogens compared with PCR when scores 1 and 2 were considered positive. However, when only score 2 was considered positive, PCR and qPCR showed better agreement. Conclusions: qPCR demonstrated higher sensitivity than conventional PCR for detection of low numbers of microorganisms and can be useful for the quantification of periodontopathogens.
https://doi.org/10.20396/bjos.v15i3.8649599
PDF (Português (Brasil))

References

Krishnan K, Chen T, Paster BJ. A practical guide to the oral microbiome and its relation to health and disease. Oral Dis. 2017 Apr;23(3):276-286. doi: 10.1111/odi.12509.

Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in subgingival plaque. J Clin Periodontol. 1998 Feb;25(2):13444.

Lombardo Bedran TB, Marcantonio RA, Spin Neto R, Alves Mayer MP, Grenier D, Spolidorio LC, et al. Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study. J Oral Microbiol. 2012;4. doi: 10.3402/jom.v4i0.10123.

Cosgarea R, Bäumer A, Pretzl B, Zehaczek S, Kim TS. Comparison of two different microbiological test kits for detection of periodontal pathogens. Acta Odontol Scand. 2010 Mar;68(2):115-21. doi: 10.3109/00016350903514848.

Eick S, Straube A, Guentsch A, Pfister W, Jentsch H. Comparison of real-time polymerase chain reaction and DNA-strip technology in microbiological evaluation of periodontitis treatment. Diag Microbiol Infections Dis. 2011 Jan;69(1):12-20. doi: 10.1016/j. diagmicrobio.2010.08.017.

Jervøe-Storm PM, Koltzscher M, Falk W, Dörfler A, Jepsen S. Comparison of culture and real-time PCR for detection and quantification of five putative periodontopathogenic bacteria in subgingival plaque samples. J Clin Periodontol. 2005 Jul;32(7):778-83.

Bolerázska B, Mareková M, Markovská N. Trends in Laboratory Diagnostic Methods in Periodontology. Acta Medica (Hradec Kralove). 2016;59(1):3-9. doi: 10.14712/18059694.2016.47.

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 Jul;59(7):431-8.

Feng X, Zhang L, Xu L, Meng H, Lu R, Chen Z, et al. Detection of eight

Comparison of two different methods for detecting periodontal pathogenic bacteria fimA genotypes in Chinese patients with aggressive periodontitis. J Periodontol. 2014 Jan;85(1):150-9. doi: 10.1902/jop.2013.120677.

Mullis K. The unusual origin of the polymerase chain reaction. Sci Am. 1990 Apr;262(4):56-61, 64-5.

Decat E, Cosyn J, De Bruyn H, Miremadi R, Saerens B, Van Mechelen E, et al. Optimization of quantitative polymerase chain reactions for detection and quantification of eight periodontal bacterial pathogens. BMC Res Notes. 2012 Dec 2;5:664. doi: 10.1186/1756-0500-5-664.

Marín MJ, Figuero E, González I, O'Connor A, Diz P, Álvarez M, et al. Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques. Med Oral Patol Oral Cir Bucal. 2016 May 1;21(3):e276-84.

Guglielmetti MR, Rosa EF, Lourenção DS, Inoue G, Gomes EF, De Micheli G, et al. Detection and quantification of periodontal pathogens in smokers and never-smokers with chronic periodontitis by real-time polymerase chain reaction. J Periodontol. 2014 Oct;85(10):1450-7. doi: 10.1902/jop.2014.140048.

Rudney JD, Chen R, Pan Y. Endpoint quantitative PCR assays for Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. J Periodontal Res. 2003 Oct;38(5):465-70.

Tellapragada C, Eshwara VK, Acharya S, Bhat P, Kamath A, Vishwanath S, et al. Prevalence of Clinical Periodontitis and Putative Periodontal Pathogens among South Indian Pregnant Women. Int J Microbiol. 2014; 2014:420149. doi: 10.1155/2014/420149.

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 Aug;45(2):1919.

Umeda JE, Missailidis C, Longo PL, Anzai D, Wikström M, Mayer MP. Adhesion and invasion to epithelial cells by fimA genotypes of Porphyromonas gingivalis. Oral Microbiol Immunol. 2006 Dec;21(6):4159.

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 Aug;11(4):266-73.

Persson GR, Hitti J, Paul K, Hirschi R, Weibel M, Rothen M, et al. Tannerella forsythia and Pseudomonas aeruginosa in subgingival bacterial samples from parous women. J Periodontol. 2008 Mar;79(3):508-16. doi: 10.1902/ jop.2008.070350.

Lages EJ, Costa FO, Cortelli SC, Cortelli JR, Cota LO, Cyrino RM, et al. Alcohol Consumption and Periodontitis: Quantification of Periodontal Pathogens and Cytokines. J Periodontol. 2015 Sep;86(9):1058-68. doi: 10.1902/jop.2015.150087.

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 Sep;76(9):1542-9.

Martin EF, Nadkarni MA, Jacques NA, Hunter N. Quantitative microbiological study of human carious dentine by culture and Real-Time PCR: association of anaerobes with histopathological changes in chronic pulpitis. J Clin Microbiol. 2002 May;40(5):1698-704.

Shaddox LM, Walker C. Microbial testing in periodontics: value, limitations and future directions. Periodontol 2000. 2009;50:25-38. doi: 10.1111/j.1600-0757.2008.00285.x.

Döring G, Unertl K, Heininger A. Validation criteria for nucleic acid amplification techniques for bacterial infections. Clin Chem Labor Med. 2008;46(7):909-18. doi: 10.1515/CCLM.2008.152.

Higuchi R, Dollinger G, Walsh PS, Griffith R. Simultaneous amplification and detection of specific DNA sequences. Biotechnology (N Y). 1992 Apr;10(4):413-7.

The Brazilian Journal of Oral Sciences uses the Creative Commons license (CC), thus preserving the integrity of the articles in an open access environment.

Downloads

Download data is not yet available.