Assessment of intraradicular bacterial composition by terminal restriction fragment length polymorphism analysis

Autor(es)

Saito, D. ^[1] ; Marsh, T. L. ^[2] ; Cannavan, F. de Souza ^[3] ; Hoefling, J. F. ^[1] ; Goncalves, R. B. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Dept Oral Diag, Div Microbiol & Immunol, Piracicaba Dent Sch, BR-13414903 Sao Paulo - Brazil

^[2] Michigan State Univ, Ctr Microbial Ecol, Dept Microbiol & Mol Genet, E Lansing, MI 48824 - USA

^[3] Univ Sao Paulo, Lab Cell & Mol Biol, Ctr Nucl Energy Agr, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Oral Microbiology and Immunology; v. 24, n. 5, p. 369-376, OCT 2009.
• Citações Web of Science: 6

Resumo

Background: The aim of the study was to assess the bacterial community structures associated with endodontic infections using terminal restriction fragment length polymorphism (T-RFLP), and to investigate the correlation of whole community profiles with the manifestation of particular clinical features. Methods: Intraradicular samples were collected from 34 subjects and classified into three study groups based on the observed clinical symptoms: acute (n = 16), sub-acute (n = 8), and asymptomatic (n = 10). Genomic DNA was extracted from each sample, submitted to polymerase chain reaction using a fluorescently labeled 16S ribosomal DNA forward primer, and digested with two tetrameric endonucleases (HhaI and MspI). The terminal restriction fragments (T-RFs) were subsequently discriminated in an automated DNA sequencer, and the results were filtered using a statistics-based criterion. Results: Totals of 138 (HhaI) and 145 (MspI) unique T-RFs were detected (means 13.1 and 11.9) and there was high inter-subject variability in the bacterial assemblages. Odds-ratio analysis unveiled the existence of higher order groups of positively associated T-RFs, restating the concept that intricate ecological relationships may take place in the root canal space. A significantly greater T-RF prevalence was detected in acute cases, suggesting a straight correlation between species richness and spontaneous pain. Conclusion: Overall, no T-RFLP profile representing a specific bacterial consortium could be associated with the manifestation of symptoms of endodontic origin. (AU)

• Processo FAPESP: 04/01674-6 - Detecção quantitativa de Filifactor alocis e Treponema socranskii em infecções endodônticas, através de PCR em tempo real
• Beneficiário: Daniel Saito
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto