Endodontik Mikrobiyolojide Klasik ve Güncel Tanı Yöntemleri
Yıl 2025,
Cilt: 4 Sayı: 1, 34 - 45, 27.03.2025
Eda Hasgül
,
Fahriye Keskin
,
Sevgi Çiftçi
,
Bülent Yılmaz
Öz
Endodontik enfeksiyonlar diş pulpasının mikroorganizmalarca istilası sonucu oluşmaktadır. Mikroorganizmalar veya toksinleri öncelikle pulpaya ulaşarak pulpitislere, ardından periapikal bölgeyi etkileyerek periapikal doku hastalıklarına neden olmaktadır. Polimikrobiyal karakterde olan endodontik enfeksiyonlarda anaerop bakteriler baskın durumdadır. Bununla birlikte, kök kanallarında bakteriler dışında mantar, virüs ve arkealar da tanımlanmıştır. Kök kanallarındaki mikroorganizma çeşitliliğinin anlaşılması enfeksiyonun bütünüyle yok edilmesi, yeniden oluşabilecek bir enfeksiyonun önlenmesi ve tedaviye dirençli olgularda primer enfeksiyon etkeni dışındaki mikroorganizmaların tespiti noktasında fayda sağlamaktadır. Dolayısıyla, endodontik tedavinin başarısı, endodontik uygulamaların mikrobiyolojik çalışmalarla birlikte yürütülmesi ile artmaktadır. Günümüzde, endodontik mikrobiyolojide kültür yöntemi, enfeksiyon etkenlerinin saptanmasında hala standart tanı yöntemi olarak kullanılmaya devam etmektedir. Son yıllarda ise mikrobiyatanın çeşitliliğini belirleyebilmek için moleküler tanı yöntemleri ön plana çıkmaktadır. Bu makalenin amacı klinisyenlere endodonti alanında kullanılan mikrobiyolojik tanı yöntemlerini tanıtmak ve bu yöntemlerin avantaj ve dezavantajlarını ortaya koymaktır.
Kaynakça
- Ahlat, M., Aydın, C., Kaya, S. & Baysallar, M. (2023). "Identification of Root Canal Microbiota Profiles of Periapical Tissue Diseases Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry." Anaerobe 84: 1027-91.
- Arana-Chavez, V. E. & Castro-Filice, L. S. (2019). "Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) for the Examination of Dental Hard Tissues." Methods in Molecular Biology, 1922: 325-332.
- Arcuri, L., Gambarini, G., Zanza, A., Testarelli, L., Arcuri, C., Cross, R., et al. (2024). "Experimental Evaluation of a Novel Device to Quantify Canal Cleanliness: An In Vitro Study." ORAL, 4(2), 163-172.
- Bagg, J., MacFarlane, T.W., Poxton, I.R. & Smith, A.J. (2005). Essentials of Microbiology for Dental Students. Oxford: Oxford Academic Press, 127-134.
- Barbuddhe, S.B., Maier, T., Schwarz, G., Kostrzewa, M., Hof, H., Domann, E., et al., (2008). "Rapid Identification and Typing of Listeria Species by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry." Applied and Environmental Microbiology, 74/17: 5402.
- Baumgartner, J., (2004). "Microbiological and Molecular Analysis of Endodontic Infections." Endodontic Topics, 7: 35-51.
- Belibasakis, G. N., Belstrøm, D., Eick, S., Gursoy, U. K., Johansson, A. & Könönen, E. (2023). "Periodontal Microbiology and Microbial Etiology of Periodontal Diseases: Historical Concepts and Contemporary Perspectives." Periodontology 2000.
- Bergmans, L., Moisiadis, P., Van Meerbeek, B., Quirynen, M. & Lambrecht, P. (2005). "Microscopic Observation of Bacteria: Review Highlighting the Use of Environmental SEM." International Endodontic Journal, 38: 775-88.
- BioMérieux. (2021). ETEST®. [çevrimiçi] www.biomerieux-usa.com. [Erişim tarihi: 23 Ekim 2024].
- Brown, T. A. (2010). Gene Cloning and DNA Analysis: An Introduction. 5th ed. Manchester: Blackwell Publishing.
- Buonavoglia, A., Zamparini, F., Lanave, G., Pellegrini, F., Diakoudi, G., Spinelli, A., et al. (2023). "Endodontic Microbial Communities in Apical Periodontitis." Journal of Endodontics, 49/2: 178-189.
- Byrne, S., Butler, C. A., Reynolds, E. C. & Dashper, S. G. (2018). "Taxonomy of Oral Bacteria." In Methods in Microbiology, Volker Gurtler, Jack T. Trevors (eds.), 45: 171-201. Academic Press.
- Cardoso, F. G., Chung, A., Martinho, F. C., Camargo, C. H., Carvalho, C. A., Gomes, B. P., et al. (2016). "Investigation of Bacterial Contents From Persistent Endodontic Infection and Evaluation of Their Inflammatory Potential." Braz Dent J., 27/4: 412-418.
- Characklis, W. G. & Marshall, K. C. (1990). Biofilms: A Basis for an Interdisciplinary Approach. New York: John Wiley and Sons, 3-15.
- Claesson, R., Johansson, A., & Belibasakis, G. N. (2022). "Clinical Laboratory Diagnostics in Dentistry: Application of Microbiological Methods." Front Oral Health, 8/3: 983991.
- Engvall, E., & Perlmann, P. (1971). "Enzyme-linked Immunosorbent Assay (ELISA): Quantitative Assay of Immunoglobulin G." Immunochemistry, 8: 871-879.
- EUCAST (European Committee on Antimicrobial Susceptibility Testing) (2021). "Antimicrobial Susceptibility Testing: EUCAST Disk Diffusion Method." [çevrimiçi] www.eucast.org [Erişim tarihi: 23 Ekim 2024].
- Fouad, A. F., Barry, J., Caimano, M., Clawson, M., Zhu, Q., Carver, R., et al. (2002). "PCR-based Identification of Bacteria Associated with Endodontic Infections." Journal of Clinical Microbiology, 40: 3223-3231.
- Fouad, A. F. (2017). Endodontic Microbiology Second Edition. USA: John Wiley & Sons Inc.
- Fraher, M. H., O'Toole, P. W., & Quigley, E. M. (2012). "Techniques Used to Characterize the Gut Microbiota: A Guide for the Clinician." Nat Rev Gastroenterol Hepatol., 9/6: 312-322.
- Francisco, P. A., Delboni, M. G., Lima, A. R., Xiao, Y., Siqueira, W. L., & Gomes, B. (2019). "Proteomic Profile of Root Canal Contents in Teeth with Post-Treatment Endodontic Disease." Int Endod J., 52: 451-460.
- Fredricks, D. N., & Relman, D. A. (1999). "Application of Polymerase Chain Reaction to the Diagnosis of Infectious Diseases." Clin Infect Dis, 29: 475-486.
- Garg, A., Mala, K., & Kamath, P. M. (2021). "Biofilm Models in Endodontics - A Narrative Review." J Conserv Dent, 24/1: 2-9.
- Hidayat, R., & Wulandari, P. (2012). "Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline." Bioscientia Medicina: Journal of Biomedicine and Translational Research, 5/5: 447-453.
- Hoglund Aberg, C., Kwamin, F., Claesson, R., Dahlen, G., Johansson, A., & Haubek, D. (2014). "Progression of Attachment Loss is Strongly Associated with Presence of the JP2 Genotype of Aggregatibacter actinomycetemcomitans: A Prospective Cohort Study of a Young Adolescent Population." J Clin Periodontol, 41/3: 232-241.
- Hornbeck, P. (2015). "Enzyme-linked Immunosorbent Assays." Curr. Protoc. Immunol., 110: 211-2123.
Im, K., Mareninov, S., Diaz, M. F. P., & Yong, W. H. (2019). "An Introduction to Performing Immunofluorescence Staining." Methods Mol Biol., 1897: 299-311.
- Kishen, A., George, S., & Kumar, R. (2006). "Enterococcus faecalis-Mediated Biomineralized Biofilm Formation on Root Canal Dentine In Vitro." J Biomed Mater Res A, 77: 406-415.
- Korona-Glowniak, I., Piatek, D., Fornal, E., Lukowiak, A., Gerasymchuk, Y., Kedziora, A., et al. (2021). "Patterns of Oral Microbiota in Patients with Apical Periodontitis." J Clin Med., 10/12: 2707.
- Kunin, V., Engelbrektson, A., Ochman, H., & Hugenholtz, P. (2010). "Wrinkles in the Rare Biosphere: Pyrosequencing Errors Can Lead to Artificial Inflation of Diversity Estimates." Environmental Microbiology, 12: 118-123.
- Leavitt, J., Naidorf, I., & Shugaevsky, P. (1958). "The Bacterial Flora of Root Canals as Disclosed by a Culture Medium for Endodontics." Oral Surg, 11: 302-306.
- Leeuwenhoek, A. (1684). "Leeuwenhoek’s Letter." Philos Trans R Soc London, 14: 568-574.
- Lewis, K. (2001). "Riddle of Biofilm Resistance." Antimicrob Agents Chemother, 45: 999-1007.
- Lübeck, P. S., & Hoorfar, J. (2003). "PCR Technology and Applications to Zoonotic Food-Borne Bacterial Pathogens." Methods Mol Biol., 216: 65-84.
- Lyons, S. R., Griffen, A. L., & Leys, E. J. (2000). "Quantitative Real-Time PCR for Porphyromonas gingivalis and Total Bacteria." J Clin Microbiol, 38/6: 2362-2365.
- Mackay, I. M., Arden, K. E., & Nitsche, A., (2002). "Real-Time PCR in Virology." Nucleic Acids Res., 30/6: 1292-1305.
- Mandlik, J., Shah, N., Sharma, A., & Desai, M. (2016). "Microbial Identification in Endodontic Infections with an Emphasis on Molecular Diagnostic Methods: A Review." IIOAB J, 7: 60-70.
- Martinho, F. C., Leite, F. R., Nascimento, G. G., Cirelli, J. A., & Gomes, B. P. (2014). "Clinical Investigation of Bacterial Species and Endotoxin in Endodontic Infection and Evaluation of Root Canal Content Activity Against Macrophages by Cytokine Production." Clin Oral Investig., 18/9: 2095-2102.
- Matoso, F. B., Montagner, F., Grecca, F. S., Rampelotto, P. H., & Kopper, P. M. P. (2024). "Microbial Composition and Diversity in Intraradicular Biofilm Formed In Situ: New Concepts Based on Next-Generation Sequencing." Mol Oral Microbiol.
- Mellmann, A., Cloud, J., Maier, T., Keckevoet, U., Ramminger, I., Iwen, P., et al., (2008). "Evaluation of Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry in Comparison to 16S rRNA Gene Sequencing for Species Identification of Nonfermenting Bacteria." J Clin Microbiol, 46/6: 1946-1954.
- Miller, W. (1894). "An Introduction in the Study of the Bacteriopathology of the Dental Pulp." Dent Cosmos, 36: 505.
Mohammadi, Z., Palazzi, F., Giardino, L., & Shalavi, S. (2013). "Microbial Biofilms in Endodontic Infections: An Update Review." Biomed J., 36/2: 59-70.
- Mullis, K. B., (1990). "The Unusual Origin of the Polymerase Chain Reaction." Scientific American, 262/4: 56-65.
Nandakumar, R., Madayiputhiya, N., & Fouad, A. F. (2009). "Proteomic Analysis of Endodontic Infections by Liquid Chromatography-Tandem Mass Spectrometry." Oral Microbiol Immunol, 24: 347-352.
- Olsen, I., Preza, D., Aas, A. J., & Paster, B. J. (2009). "Cultivated and Not-Yet-Cultivated Bacteria in Oral Biofilms." Microbial Ecology in Health and Disease Informa, 21/2: 65-71.
- Ordinola-Zapata, R., Costalonga, M., Nixdorf, D., Dietz, M., Schuweiler, D., Lima, B. P., et al. (2023). "Taxonomic Abundance in Primary and Secondary Root Canal Infections." Int Endod J, 56/2: 278-288.
- Osborn, A. M., Moore, E. R., & Timmis, K. N. (2000). "An Evaluation of Terminal-Restriction Fragment Length Polymorphism (T-RFLP) Analysis for the Study of Microbial Community Structure and Dynamics." Environmental Microbiology, 2/1: 39-50.
- Pinheiro, E. T., Gomes, B. P. F. A., Drucker, D. B., Zaia, A. A., Ferraz, C. C. R., & Souza-Filho, F. J, (2004). "Antimicrobial Susceptibility of Enterococcus faecalis Isolated from Canals of Root Filled Teeth with Periapical Lesions." Int Endod J, 37/11: 756-763.
- Pinheiro, E. T., Karygianni, L., Candeiro, G. T. M., Vilela, B. G., Dantas, L. O., Pereira, A. C. C., et al., (2024). "Metatranscriptome and Resistome of the Endodontic Microbiome." J Endod, 50/8: 1059-1072.
- Pinto, K. P., Barbosa, A. F. A., Silva, E. J. N. L., Santos, A. P. P., & Sassone, L. M. (2023). "What Is the Microbial Profile in Persistent Endodontic Infections? A Scoping Review." J Endod, 49/7: 786-798.
- Premjeet, S., Deepika, G., Sudeep, B., Sonam, J., Sahil, K., Devashish, R., et al. (2011). "Enzyme-Linked Immuno-Sorbent Assay (ELISA), Basics and Its Application: A Comprehensive Review." J. Pharm. Res, 4/12: 4581-4583.
- Provenzano, J. C., Siqueira, J. F. Jr, Rôças, I. N., Domingues, R. R., Paes Leme, A. F., & Silva, M. R. (2013). "Metaproteome Analysis of Endodontic Infections in Association with Different Clinical Conditions." Plos One, 8: 76-108.
- Rôças, I. N., & Siqueira, J. F. Jr, (2018). "Frequency and Levels of Candidate Endodontic Pathogens in Acute Apical Abscesses as Compared to Asymptomatic Apical Periodontitis." PLoS One, 13: 0190469.
- Ronaghi, M., Karamohamed, S., Pettersson, B., Uhlen, M., & Nyren, P. (1996). "Real-Time DNA Sequencing Using Detection of Pyrophosphate Release." Anal Biochem, 242/1: 84-89.
- Rosa, T. P., Signoretti, F. G., Montagner, F., Gomes, B. P., & Jacinto, R. C. (2015). "Prevalence of Treponema spp. in Endodontic Retreatment-Resistant Periapical Lesions." Braz Oral Res, 29: 6-18.
- Sahiner, F. (2014). "Current Problems and Recent Advances in the Molecular Diagnosis of Genital Human Papillomavirus Infections." Mikrobiyol Bul, 48/4: 689-706.
- Singh, S. (2022). "Microscopes in Conservative Dentistry and Endodontics Research." J Conserv Dent, 25/4: 333-337.
- Singh, S., & Bolla, N. (2024). "Endocator-Paradigm Shift in the Evaluation of Microbial Load in Root Canals." J Conserv Dent Endod, 27: 111-112.
- Siqueira, J. F. Jr. (2002). "Endodontic Infections: Concepts, Paradigms, and Perspectives." Oral Surgery Oral Medicine Oral Pathology Oral Radiology and Endodontology, 94: 281-293.
- Siqueira, J. F. Jr., & Rôças, I. N. (2003). "Treponema socranskii in Primary Endodontic Infections as Detected by Nested PCR." J Endod, 29/4: 244-247.
- Siqueira, J. F. Jr., & Rôças, I. N. (2005). "Exploiting Molecular Methods to Explore Endodontic Infections: Part 1–Current Molecular Technologies for Microbiological Diagnosis." Journal of Endodontics, 31: 411-423.
- Socransky, S. S., Smith, C., Martin, L., Paster, B. J., Dewhirst, F. E., & Levin, A. E. (1994). "Checkerboard DNA-DNA Hybridization." Biotechniques, 17: 788-792.
- Sunde, P. T., Olsen, I., Göbel, U. B., Theegarten, D., Winter, S., Debelian, G. J., et al. (2003). "Fluorescence in Situ Hybridization (FISH) for Direct Visualization of Bacteria in Periapical Lesions of Asymptomatic Root-Filled Teeth." Microbiology (Reading), 149/5: 1095-1102.
- Tandjung, L., Waltimo, T., Hauser, I., Heide, P., Decker, E. M., & Weiger, R. (2007). "Octenidine in Root Canal and Dentine Disinfection Ex Vivo." International Endodontic Journal, 40: 845-851.
- Teles, A. M., Manso, M. C., Pina, C., & Cabeda, J. (2013). "A Review of Microbiological Root Canal Sampling: Updating an Emerging Picture." Archives of Oral Research, 9/1: 41-59.
- Trope, M., Rosenberg, E., & Tronstad, L. (1992). "Darkfield Microscopic Spirochete Count in the Differentiation of Endodontic and Periodontal Abscesses." Journal of Endodontics, 18: 82–86.
- Unnikrishnan, M. (2024). "Molecular Diagnostic Methods in Endodontics." Acta Scientific Dental Sciences, 8: 115–119.
Vauterin, L. (2006). Molecular Identification, Systematics and Population Structure of Prokaryotes. Berlin: Springer-Verlag Berlin Heidelberg, 23–50.
- Ruelle, V., El Moualij, B., Zorzi, W., Ledent, P., & Pauw, E. D. (2004). "Rapid Identification of Environmental Bacterial Strains by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry." Rapid Communications in Mass Spectrometry, 18: 2013–2019.
- Wong, J., Manoil, D., Näsman, P., Belibasakis, G. N., & Neelakantan, P. (2021). "Microbiological Aspects of Root Canal Infections and Disinfection Strategies: An Update Review on the Current Knowledge and Challenges." Front Oral Health, 2: 672887.
- Yolcu, E. N., Tartuk, G., Kaya, S., & Eskibağlar, M. (2021). "The Use of Confocal Laser Scanning Microscopy in Endodontics: A Literature Review." Turk Endod J, 6(2): 55-60.
- Zapata, R. O., Bramante, C. M., de Moraes, I. G., Bernardineli, N., Gasparoto, T. H., Graeff, M. S., et al. (2008). "Confocal Laser Scanning Microscopy Is Appropriate to Detect Viability of Enterococcus faecalis in Infected Dentin." J Endod, 34: 1198–201.