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

• Published in: Journal of endodontics Vol. 43; no. 3; pp. 377 - 382
• Main Authors: Delboni, Maraísa G., DDS, MSc, PhD, Gomes, Brenda P.F.A., DDS, MSc, PhD, Francisco, Priscila A., DDS, Teixeira, Fabrício B., DDS, MSc, PhD, Drake, David, DDS, MSc, PhD
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2017
• Subjects: Arbitrarily primed polymerase chain reaction; Dental Pulp Cavity - microbiology; Dentistry; DNA Fingerprinting; DNA, Bacterial - analysis; Endocrinology & Metabolism; endodontics; Enterococcus faecalis; Enterococcus faecalis - genetics; Enterococcus faecalis - isolation & purification; Genotype; Humans; microleakage; microorganisms; Periapical Periodontitis - microbiology; Polymerase Chain Reaction - methods; repetitive sequence-based polymerase chain reaction; Repetitive Sequences, Nucleic Acid; saliva; Saliva - microbiology; Tooth, Nonvital - microbiology; Treatment Failure; endodontics; Enterococcus faecalis; saliva; microorganisms; repetitive sequence-based polymerase chain reaction; Arbitrarily primed polymerase chain reaction; microleakage
• ISSN: 0099-2399; 1878-3554
• DOI: 10.1016/j.joen.2016.10.042

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.