Identification of microorganisms in persistent/secondary endodontic infections with respect to clinical and radiographic findings: bacterial culture and molecular detection
,
Abstract
Background and Objectives: Bacterial agents are commonly accepted as the main etiology of endodontic infections. A significant proportion of oral bacteria cannot be cultured using existing methods. Since diversity and abundance of bacterial species are different in different populations, the present study was aimed to identify effective microorganisms in persistent endodontic infections in Iranian patients based on culture and molecular biology methods using sequence analysis of 16S rDNA gene.
Materials and Methods: Thirty patients with previous failure of endodontic treatment were enrolled in the study. After isolation and disinfection of the tooth surrounding area with 3% sodium hypochlorite and 30% hydrogen peroxide, sampling from the root canals was carried out using two sterile Hedstrom files and two sterile paper points, and then the specimens were transferred to the microbiology laboratory in thioglycolate transport medium so that they undergo aerobic-anaerobic culture, PCR, and 16S rDNA gene sequencing.
Results: Of 30 patients (15 women and 15 men), 15 patients had radiographic lesions smaller than 5 mm and other 15 patients had radiographic lesions larger than 5 mm. The mean age of patients was 40.20 ± 13.76 years. A total of 26 patients were asymptomatic. Only four patients had clinical signs such as pain and percussion sensitivity and Tannerella forsythia was the most common bacterium found in this group of patients. 13 bacterial species were found in 11 different genus, one virus strain and one fungus strain. From 30 studied specimens, Enterococcus faecalis was the most common microorganism with prevalence rate of 63.63%.
Conclusion: This study showed the type and prevalence of effective bacteria in secondary/persistent endodontic infections in Iranian patients. E. faecalis is the most commonly found microorganism in Iranian patients.
References:
C.F. Murad, L.M. Sassone. Microbial Diversity in Persistent Root Canal Infections Investigated by Checkerboard DNA-DNA Hybridization J Endod. 2014 ;40(7):899-906
Christian Tennert, Maximilian Fuhrmann. New Bacterial Composition in Primary and Persistent/Secondary Endodontic Infections with Respect to Clinical and Radiographic Findings. J Endod 2014;40:670–677
Kakehashi S, Stanley HR, Fitzgerald RJ. The effects of surgical exposures of dental pulps in Germ-free and conventional laboratory rats. Oral Surg Oral Med Oral Pathol 1965; 20: 340-349.
Sakamoto M, Siqueira Jr JF, Rôças IN, Benno Y. Molecular analysis of the root canal microbiota associated with endodontic treatment failures. Oral microbiology and immunology. 2008 Aug 1;23(4):275-81.
Rôças IN, Siqueira JF. Characterization of microbiota of root canal-treated teeth with posttreatment disease. Journal of clinical microbiology. 2012 May 1;50(5):1721-4.
Schirrmeister JF, Liebenow AL, Pelz K, Wittmer A, Serr A, Hellwig E, Al-Ahmad A. New bacterial compositions in root-filled teeth with periradicular lesions. Journal of endodontics. 2009 Feb 28;35(2):169-74.
Anderson AC, Hellwig E, Vespermann R, Wittmer A, Schmid M, Karygianni L, Al-Ahmad A. Comprehensive analysis of secondary dental root canal infections: a combination of culture and culture-independent approaches reveals new insights. PloS one. 2012 Nov 12;7(11):e49576.
Rôças IN, Hülsmann M, Siqueira JF. Microorganisms in root canal–treated teeth from a German population. Journal of endodontics. 2008 Aug 31;34(8):926-31.
Gomes BP, Pinheiro ET, Jacinto RC, Zaia AA, Ferraz CC, Souza-Filho FJ. Microbial analysis of canals of root-filled teeth with periapical lesions using polymerase chain reaction. Journal of endodontics. 2008 May 31;34(5):537-40.
Siqueira JF, Sen BH. Fungi in endodontic infections. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2004 May 31;97(5):632-41.
Foschi F, Cavrini F, Montebugnoli L, Stashenko P, Sambri V, Prati C. Detection of bacteria in endodontic samples by polymerase chain reaction assays and association with defined clinical signs in Italian patients. Oral microbiology and immunology. 2005 Oct 1;20(5):289-95.
LI Xin, ZHU Xiao-fei, ZHANG Cheng-fei. Endodontic bacteria from primary and persistent endodontic lesions in Chinese patients as identified by cloning and 16S ribosomal DNA gene sequencing. Chin Med J 2013;126 (4): 634-639.
Zakaria MN, Takeshita T, Shibata Y, Wada N, Akamine A, Yamashita Y. Microbial community in persistent apical periodontitis: a 16S rRNA gene clone library analysis. Int Endod J. 2014 Aug 2.
Hong BY, Lee TK, Lim SM, Chang SW, Park J, Han SH, Zhu Q, Safavi KE, Fouad AF, Kum KY. Microbial analysis in primary and persistent endodontic infections by using pyrosequencing. Journal of endodontics. 2013 Sep 30;39(9):1136-40.
Anderson AC, Al-Ahmad A, Elamin F, Jonas D, Mirghani Y, Schilhabel M, Karygianni L, Hellwig E, Rehman A. Comparison of the bacterial composition and structure in symptomatic and asymptomatic endodontic infections associated with root-filled teeth using pyrosequencing. PloS one. 2013 Dec 30;8(12):e84960.
Siqueira JF Jr, Rocas IN. Uncultivated phylotypes and newly named species associated with primary and persistent endodontic infections. J Clin Microbiol 2005; 43: 3314-3319.
Siqueira JF Jr. Taxonomic changes of bacteria associated with endodontic infections. J Endod 2003; 29: 619-623.
Woo PC, Lau SK, Teng JL, Tse H, Yuen KY. Then and now: use of 16S rDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories. Clinical Microbiology and Infection. 2008 Oct 1;14(10):908-34.
Cohen's Pathways of the Pulp 11th edition, diagnosis page 27
Cohen’s pathways of the pulp, microbiology of edodontic infections page 599
LI Xin, ZHU Xiao-fei, ZHANG Cheng-fei. Endodontic bacteria from primary and persistent endodontic lesions in Chinese patients as identified by cloning and 16S ribosomal DNA gene sequencing. Chin Med J 2013;126 (4): 634-639.
Woo PC, Lau SK, Teng JL, Tse H, Yuen KY. Then and now: use of 16S rDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories. Clinical Microbiology and Infection. 2008 Oct 1;14(10):908-34.
Foundations in microbiology 9th edition, chapter 1
Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. Journal of bacteriology. 1991 Jan 1;173(2):697-703.
Wang J, Jiang Y, Chen W, Zhu C, Liang J. Bacterial flora and extraradicular biofilm associated with the apical segment of teeth with post-treatment apical periodontitis. J Endod. 2012 Jul;38(7):954-9
Rôças IN, Siqueira JF Jr. Identification of bacteria enduring endodontic treatment procedures by a combined reverse transcriptase-polymerase chain reaction and reverse-capture checkerboard approach. J Endod. 2010 Jan;36(1):45-52
Blome B, Braun A, Sobarzo V, Jepsen S. Molecular identification and quantification of bacteria from endodontic infections using real‐time polymerase chain reaction. Oral microbiology and immunology. 2008 Oct 1;23(5):384-90.
Siqueira JF, Rôças IN. Polymerase chain reaction–based analysis of microorganisms associated with failed endodontic treatment. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2004 Jan 31;97(1):85-94.
Li H, Chen V, Chen Y, Baumgartner JC, Machida CA. Herpesviruses in endodontic pathoses: association of Epstein-Barr virus with irreversible pulpitis and apical periodontitis. Journal of endodontics. 2009 Jan 31;35(1):23-9.
Sabeti M, Simon JH, Slots J. Cytomegalovirus and Epstein–Barr virus are associated with symptomatic periapical pathosis. Oral microbiology and immunology. 2003 Oct 1;18(5):327-8.
Baumgartner JC, Watts CM, Xia T. Occurrence of Candida albicans in infections of endodontic origin. Journal of endodontics. 2000 Dec 31;26(12):695-8.
Downes J, Munson M, Wade WG. Dialister invisus sp. nov., isolated from the human oral cavity. International journal of systematic and evolutionary microbiology. 2003 Nov 1;53(6):1937-40.
Kumar PS, Griffen AL, Barton JA, Paster BJ, Moeschberger ML, Leys EJ. New bacterial species associated with chronic periodontitis. Journal of dental research. 2003 May 1;82(5):338-44.
Brinig MM, Lepp PW, Ouverney CC, Armitage GC, Relman DA. Prevalence of bacteria of division TM7 in human subgingival plaque and their association with disease. Applied and environmental microbiology. 2003 Mar 1;69(3):1687-94.
Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. Journal of bacteriology. 1991 Jan 1;173(2):697-703.
https://www.ncbi.nlm.nih.gov/taxonomy
Rolph HJ, Lennon A, Riggio MP, Saunders WP, MacKenzie D, Coldero L, Bagg J. Molecular identification of microorganisms from endodontic infections. Journal of clinical microbiology. 2001 Sep 1;39(9):3282-9.
Subramanian K, Mickel AK. Molecular analysis of persistent periradicular lesions and root ends reveals a diverse microbial profile. Journal of endodontics. 2009 Jul 31;35(7):950-7.
Andersen BM, Steigerwalt AG, O'Connor SP, Hollis DG, Weyant RS, Weaver RE, Brenner DJ. Validation of the Publication of New Names and New Combinations Previously Effectively Published Outside the IJSB: List no. 47. International Journal of Systematic and Evolutionary Microbiology. 1993;43(4):864-5.
Estrela C, Silva JA, Alencar AH, Leles CR, Decurcio DA. Efficacy of sodium hypochlorite and chlorhexidine against Enterococcus faecalis: a systematic review. Journal of Applied oral science. 2008 Dec;16(6):364-8.
| Files | ||
| Issue | Vol 11 No 2 (2019) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v11i2.1073 | |
| Keywords | ||
| Bacterial identification Clinical signs Polymerase chain reaction Endodontic infection 16S rDNA Enterococcus faecalis | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
