Maraísa G Delboni1, Brenda P F A Gomes2, Priscila A Francisco3, Fabrício B Teixeira4, David Drake4. 1. College of Dentistry, Facid DeVry University, Teresina, Piauí, Brazil; Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil. 2. Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil. Electronic address: bpgomes@unicamp.br. 3. Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil. 4. Department of Endodontics, Dows Institute for Dental Research, College of Dentistry, University of Iowa, Iowa City, Iowa.
Abstract
INTRODUCTION: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
INTRODUCTION: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: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.
Authors: Priscila Amanda Francisco; Pedro Ivo da Graça Fagundes; João Carlos Lemes-Junior; Augusto Rodrigues Lima; Maicon Ricardo Zieberg Passini; Brenda P F A Gomes Journal: Clin Oral Investig Date: 2021-02-09 Impact factor: 3.573
Authors: Amjad Abu Hasna; Ana Luisa Theodoro; Larissa Marques Pereira; Lucas de Paula Ramos; Tiago Moreira Bastos Campos; Maisour Ala Rachi; Talal Al-Nahalwi; Luciane Dias de Oliveira; Cláudio Antonio Talge Carvalho Journal: Biomed Res Int Date: 2022-05-10 Impact factor: 3.246
Authors: Marlos Barbosa-Ribeiro; Rodrigo Arruda-Vasconcelos; Lidiane M Louzada; Danielle G Dos Santos; Fernando D Andreote; Brenda P F A Gomes Journal: Clin Oral Investig Date: 2020-08-28 Impact factor: 3.573