title: Phenotypic and genotypic identification of enterococci isolated from canals of root-filled teeth with periapical lesions.
abstract: The objectives of the present study were to identify enterococcal species isolated from the canals of root-filled teeth with periapical lesions using biochemical and molecular techniques, and to investigate the genetic diversity of the isolates. Twenty-two Enterococcus strains, isolated from the canals of root-filled teeth with persisting periapical lesions, were identified to species level using rapid ID 32 STREP galleries and partial 16S rDNA sequencing. To subtype the strains, genomic DNA from the isolates was analyzed by pulsed field gel electrophoresis (PFGE) after digestion with SmaI. Intragenic regions of two genes, ace and salA, were sequenced for further differentiation of the isolates. All strains were identified as Enterococcus faecalis by both commercial kit and partial 16S rDNA sequencing. PFGE with SmaI of 22 isolates demonstrated 18 macrorestriction profiles, whereas 13 distinct genotypes were identified after analysis of the ace and salA composite sequences. Most of the isolates from distinct patients had different PFGE profiles. Moreover, in two cases, different E. faecalis strains were found in different root-filled teeth from the same mouth. E. faecalis was the only enterococcal species isolated from the canals of root-filled teeth with periapical lesions. Genetic heterogeneity was observed among the E. faecalis isolates following PFGE and sequence-based typing method. Furthermore, the genetic diversity within root canal strains was similar to previous reports regarding E. faecalis isolates from different clinical and geographic origins.
authors.affiliation: Endodontic Area, Dental School of Piracicaba, University of Campinas - UNICAMP, Piracicaba, São Paulo, Brazil.
authors.forename: E T
authors.initials: ET
authors.lastname: Pinheiro
authors.forename: M J
authors.initials: MJ
authors.lastname: Anderson
authors.forename: B P F A
authors.initials: BP
authors.lastname: Gomes
authors.forename: D B
authors.initials: DB
authors.lastname: Drucker
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002352:Carrier Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D003786:Dental Pulp Cavity / Q000382:microbiology*; D016521:Electrophoresis, Gel, Pulsed-Field; D013293:Enterococcus faecalis / Q000235:genetics* / Q000302:isolation & purification; D014644:Genetic Variation; D006801:Humans; D008969:Molecular Sequence Data; D010485:Periapical Periodontitis / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D019553:Tooth, Nonvital / Q000382:microbiology*
chemical_list: C121675:Ace protein, Enterococcus; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial