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ORIGINAL RESEARCH article

Front. Oral. Health
Sec. Oral Infections and Microbes
Volume 6 - 2025 | doi: 10.3389/froh.2025.1520945
This article is part of the Research Topic Diagnosis and Therapeutic Interventions for Oral Diseases View all articles

Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment

Provisionally accepted
Gabriela Cecon Chianca Gabriela Cecon Chianca 1Caroline Corrêa da Silva Caroline Corrêa da Silva 1Sérgio Pinto Chaves Junior Sérgio Pinto Chaves Junior 2Gabriella Lorena Dias Pereira Gabriella Lorena Dias Pereira 1Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa 1Lívia Antunes Lívia Antunes 1Leonardo Antunes Leonardo Antunes 1Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio 1*
  • 1 Universidade Federal Fluminense, Nova Friburgo, Brazil
  • 2 Fluminense Federal University, Niterói, Brazil

The final, formatted version of the article will be published soon.

    Microorganisms have an important role in the pathogenesis of endodontic disease. Significant advances have been made to increase the sensitivity of microbial detection, identification and enumeration in endodontic samples. The aim of the present study is to compare culture and whole-genome amplification (WGA) followed by PCR assays in the detection of bacteria before and after chemical mechanical preparation (CMP) of root canals. Ten uniradicular teeth with primary endodontic infections were analyzed. Microbiological samples were collected before and after CMP using paper points, which were separated into two groups: (i) culture assay samples were plated onto Brucella agar with 5% defibrinated sheep's blood, menadione and hemin and incubated anaerobically for 14 days at 36°C; (ii) DNA was extracted from molecular assay samples and subject to WGA by isothermal strand displacement with Phi29 DNA polymerase followed by PCR to determine the presence of bacteria. In both assays, samples before CMP showed the presence of bacteria in all 10 teeth. After CMP, however, bacterial detection differed in the assays performed (p = 0.0198). The presence of bacteria was detected in 70% (7 of 10) of the samples by WGA followed by PCR, where only 10% (1 of 10) had demonstrated bacterial growth in the culture method. The combination of WGA followed by PCR increased the detection of microorganisms from root canal samples after endodontic treatment using NaOCl as a CMP irrigant. So this combination of techniques can represent an important tool to improve the detection of microorganisms in endodontic research.

    Keywords: Dental Pulp Necrosis, Microbiology, Nucleic Acid Amplification Techniques, Polymerase Chain Reaction, Bacteria

    Received: 31 Oct 2024; Accepted: 13 Jan 2025.

    Copyright: © 2025 Chianca, da Silva, Junior, Pereira, Póvoa, Antunes, Antunes and Iorio. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Natalia Lopes Pontes Póvoa Iorio, Universidade Federal Fluminense, Nova Friburgo, Brazil

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.