Molecular analysis of the root canal microbiota associated with endodontic treatment failures.

INTRODUCTION The failure of endodontic treatment is usually caused by persistent/secondary intraradicular infections and Enterococcus faecalis has been considered to be the main pathogen involved. Nevertheless, the breadth of bacterial diversity involved with endodontic treatment failures remains to be consistently explored by culture-independent approaches. METHODS This study determined the intraradicular microbiota of root-canal-treated teeth with post-treatment apical periodontitis using 16S ribosomal RNA gene clone library analysis. RESULTS Bacteria were present in all cases, confirming the infectious etiology of post-treatment disease. Seventy-four bacterial taxa belonging to six phyla were found in the nine cases investigated. Of these, 55% were identified as as-yet-uncultivated phylotypes, which also made up a significant proportion of the microbiota in many cases. Twenty-five new phylotypes were identified. Most teeth harbored a mixed consortium, with a mean number of 10 taxa per case. Only 11 taxa were found in more than one case, revealing a high interindividual variability in the composition of the microbiota. CONCLUSION The current findings revealed new candidate endodontic pathogens, including as-yet-uncultivated bacteria and taxa other than E. faecalis, which may participate in the mixed infections associated with post-treatment apical periodontitis.

[1]  B. Gomes,et al.  Microorganisms from canals of root-filled teeth with periapical lesions. , 2003, International endodontic journal.

[2]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[3]  M. Sakamoto,et al.  Molecular analysis of bacteria in asymptomatic and symptomatic endodontic infections. , 2006, Oral microbiology and immunology.

[4]  M. Haapasalo,et al.  Enterococcus faecalis– the root canal survivor and ‘star’ in post-treatment disease , 2003 .

[5]  D. Lane 16S/23S rRNA sequencing , 1991 .

[6]  I. N. Rôças,et al.  Exploiting molecular methods to explore endodontic infections: Part 1--current molecular technologies for microbiological diagnosis. , 2005, Journal of endodontics.

[7]  J F Siqueira,et al.  Aetiology of root canal treatment failure: why well-treated teeth can fail. , 2001, International endodontic journal.

[8]  Sudhir Kumar,et al.  MEGA2: molecular evolutionary genetics analysis software , 2001, Bioinform..

[9]  James R. Cole,et al.  The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy , 2003, Nucleic Acids Res..

[10]  A. Lennon,et al.  Molecular Identification of Microorganisms from Endodontic Infections , 2001, Journal of Clinical Microbiology.

[11]  W. Wade,et al.  Molecular and Cultural Analysis of the Microflora Associated with Endodontic Infections , 2002 .

[12]  J. A. Aas,et al.  Defining the Normal Bacterial Flora of the Oral Cavity , 2005, Journal of Clinical Microbiology.

[13]  U. Göbel,et al.  Fluorescence in situ hybridization (FISH) for direct visualization of bacteria in periapical lesions of asymptomatic root-filled teeth. , 2003, Microbiology.

[14]  I. N. Rôças,et al.  Uncultivated Phylotypes and Newly Named Species Associated with Primary and Persistent Endodontic Infections , 2005, Journal of Clinical Microbiology.

[15]  Renato de Toledo Leonardo,et al.  Identification of bacteria in endodontic infections by sequence analysis of 16S rDNA clone libraries. , 2006, Journal of medical microbiology.

[16]  D. Figdor Apical periodontitis: a very prevalent problem. , 2002, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[17]  F. Dewhirst,et al.  Bacterial Diversity in Human Subgingival Plaque , 2001, Journal of bacteriology.

[18]  J. Siqueira,et al.  Identification of Enterococcus faecalis in root-filled teeth with or without periradicular lesions by culture-dependent and-independent approaches. , 2006, Journal of endodontics.

[19]  I. N. Rôças,et al.  Polymerase chain reaction identification of microorganisms in previously root-filled teeth in a South Korean population. , 2004, Journal of endodontics.

[20]  M. Moeschberger,et al.  Identification of Candidate Periodontal Pathogens and Beneficial Species by Quantitative 16S Clonal Analysis , 2005, Journal of Clinical Microbiology.

[21]  B. Kaufman,et al.  Enterococcus spp. in endodontically treated teeth with and without periradicular lesions. , 2005, Journal of endodontics.

[22]  M. Moeschberger,et al.  Molecular Analysis of Bacterial Species Associated with Childhood Caries , 2002, Journal of Clinical Microbiology.

[23]  A. Rosado,et al.  Denaturing gradient gel electrophoresis analysis of bacterial communities associated with failed endodontic treatment. , 2004, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[24]  M. Moeschberger,et al.  Changes in Periodontal Health Status Are Associated with Bacterial Community Shifts as Assessed by Quantitative 16S Cloning and Sequencing , 2006, Journal of Clinical Microbiology.

[25]  C. Reit,et al.  Microbiological status of root-filled teeth with apical periodontitis. , 1998, International endodontic journal.

[26]  U Sjögren,et al.  Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment. , 1998, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[27]  F. Dewhirst,et al.  Diversity of Bacterial Populations on the Tongue Dorsa of Patients with Halitosis and Healthy Patients , 2003, Journal of Clinical Microbiology.

[28]  J. A. Aas,et al.  The breadth of bacterial diversity in the human periodontal pocket and other oral sites. , 2006, Periodontology 2000.

[29]  I. N. Rôças,et al.  Polymerase chain reaction-based analysis of microorganisms associated with failed endodontic treatment. , 2004, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.