Journal article
Diversity of Enterococcus faecalis Genotypes from Multiple Oral Sites Associated with Endodontic Failure Using Repetitive Sequence-based Polymerase Chain Reaction and Arbitrarily Primed Polymerase Chain Reaction
Journal of endodontics, Vol.43(3), pp.377-382
03/2017
DOI: 10.1016/j.joen.2016.10.042
PMID: 28131414
Abstract
The aim of this study was to evaluate the diversity and similarity of Enterococcus faecalis genotype isolates from multiple oral sites using repetitive sequence-based polymerase chain reaction and arbitrarily primed polymerase chain reaction (AP-PCR).
Forty-two endodontically treated teeth with apical periodontitis were selected. A total of 126 microbial samples were collected from 3 different sites (saliva, pulp chamber, and root canals, all n = 42) during the nonsurgical retreatment procedures. After growth on m-Enterococcus agar, the colonies were isolated, characterized as gram-positive catalase negative cocci, and identified using an API 20 Strep kit (bioMérieux, Marcy-l'Etoile, France). Seventy-four colonies from 10 patients were confirmed as E. faecalis by polymerase chain reaction (16S ribosomal RNA). Repetitive sequence-based polymerase chain reactions using ERIC and AP-PCR using RW3A primers were performed in all 74 colonies. Fingerprints were analyzed and separated into genotypic groups based on the Dice coefficient percentage of similarity (82% or greater) as determined by ERIC reproducibility assays involving E. faecalis controls.
Seven different E. faecalis genotypes (GTs) (GT1 = 27%, GT2 = 17.6%, GT3 = 1.3%, GT4 = 18.9%, GT5 = 9.5%, GT6 = 14.9%, and GT7 = 10.8%) were observed in different subjects and oral sites associated with endodontic failure. Remarkably, in 4 of 5 patients, the same GTs present in the infected root canals were also isolated from either the pulp chamber or the saliva samples. In particular, GT6 was detected in all 3 oral sites of patient 37.
E. faecalis GTs isolated from saliva, the pulp chamber, and the root canal were similar using the Rep-PCR and AP-PCR methods. These findings suggest that coronal microleakage is a conceivable cause of endodontic failure.
•Tracking the pathways of the root canal reinfection.•DNA fingerprints from samples from saliva, pulp chamber and root canal.•Determination of diversity and similarity of E. faecalis genotypes.
Details
- Title: Subtitle
- Diversity of Enterococcus faecalis Genotypes from Multiple Oral Sites Associated with Endodontic Failure Using Repetitive Sequence-based Polymerase Chain Reaction and Arbitrarily Primed Polymerase Chain Reaction
- Creators
- Maraísa G Delboni - College of Dentistry, Facid DeVry University, Teresina, Piauí, BrazilBrenda P.F.A Gomes - Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, BrazilPriscila A Francisco - Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, BrazilFabrício B Teixeira - Department of Endodontics, Dows Institute for Dental Research, College of Dentistry, University of Iowa, Iowa City, IowaDavid Drake - Department of Endodontics, Dows Institute for Dental Research, College of Dentistry, University of Iowa, Iowa City, Iowa
- Resource Type
- Journal article
- Publication Details
- Journal of endodontics, Vol.43(3), pp.377-382
- DOI
- 10.1016/j.joen.2016.10.042
- PMID
- 28131414
- NLM abbreviation
- J Endod
- ISSN
- 0099-2399
- eISSN
- 1878-3554
- Publisher
- Elsevier Inc
- Grant note
- DOI: 10.13039/501100001807, name: FAPESP, award: 2015/19215-2; DOI: 10.13039/501100003593, name: CNPq, award: 302575/2009-0, 308162/2014-5; DOI: 10.13039/501100002322, name: CAPES
- Language
- English
- Date published
- 03/2017
- Academic Unit
- Endodontics; Dental Research
- Record Identifier
- 9984065996502771
Metrics
14 Record Views