Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment
IntroductionMicroorganisms 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-geno...
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Frontiers Media S.A.
2025-02-01
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| author | Gabriela Ceccon Chianca Gabriela Ceccon Chianca Caroline Corrêa Fendeler Caroline Corrêa Fendeler Sérgio Pinto Chaves Junior Sérgio Pinto Chaves Junior Gabriella Lorena Dias Pereira Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Leonardo Santos Antunes Leonardo Santos Antunes Leonardo Santos Antunes Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio |
| author_facet | Gabriela Ceccon Chianca Gabriela Ceccon Chianca Caroline Corrêa Fendeler Caroline Corrêa Fendeler Sérgio Pinto Chaves Junior Sérgio Pinto Chaves Junior Gabriella Lorena Dias Pereira Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Leonardo Santos Antunes Leonardo Santos Antunes Leonardo Santos Antunes Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio |
| author_sort | Gabriela Ceccon Chianca |
| collection | DOAJ |
| description | IntroductionMicroorganisms 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.MethodsTen 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.ResultsIn 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.ConclusionThe 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. |
| format | Article |
| id | doaj-art-b4e8412ed6b2443a9a09a36c2d73fbad |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Oral Health |
| spelling | doaj-art-b4e8412ed6b2443a9a09a36c2d73fbad2025-02-03T06:33:27ZengFrontiers Media S.A.Frontiers in Oral Health2673-48422025-02-01610.3389/froh.2025.15209451520945Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatmentGabriela Ceccon Chianca0Gabriela Ceccon Chianca1Caroline Corrêa Fendeler2Caroline Corrêa Fendeler3Sérgio Pinto Chaves Junior4Sérgio Pinto Chaves Junior5Gabriella Lorena Dias Pereira6Helvécio Cardoso Corrêa Póvoa7Helvécio Cardoso Corrêa Póvoa8Helvécio Cardoso Corrêa Póvoa9Lívia Azeredo Alves Antunes10Lívia Azeredo Alves Antunes11Lívia Azeredo Alves Antunes12Leonardo Santos Antunes13Leonardo Santos Antunes14Leonardo Santos Antunes15Natalia Lopes Pontes Póvoa Iorio16Natalia Lopes Pontes Póvoa Iorio17Natalia Lopes Pontes Póvoa Iorio18Experimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Pathology, School of Medicine, UFF, Niterói, Rio de Janeiro, BrazilExperimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Dentistry, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilExperimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Applied Microbiology and Parasitology, Biomedical Institute, UFF, Niterói, Rio de Janeiro, BrazilExperimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilExperimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Dentistry, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Applied Microbiology and Parasitology, Biomedical Institute, UFF, Niterói, Rio de Janeiro, BrazilGraduate Program in Dentistry, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilDepartment of Specific Formation, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Dentistry, School of Dentistry, UFF, Niterói, Rio de Janeiro, BrazilGraduate Program in Dentistry, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilDepartment of Specific Formation, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Dentistry, School of Dentistry, UFF, Niterói, Rio de Janeiro, BrazilExperimental and Applied Microbiology Laboratory, Nova Friburgo Institute of Health, Universidade Federal Fluminense (UFF), Nova Friburgo, Rio de Janeiro, BrazilGraduate Program in Pathology, School of Medicine, UFF, Niterói, Rio de Janeiro, BrazilGraduate Program in Dentistry, Nova Friburgo Institute of Health, UFF, Nova Friburgo, Rio de Janeiro, BrazilIntroductionMicroorganisms 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.MethodsTen 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.ResultsIn 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.ConclusionThe 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.https://www.frontiersin.org/articles/10.3389/froh.2025.1520945/fulldental pulp necrosismicrobiologynucleic acid amplification techniquespolymerase chain reactionbacteria |
| spellingShingle | Gabriela Ceccon Chianca Gabriela Ceccon Chianca Caroline Corrêa Fendeler Caroline Corrêa Fendeler Sérgio Pinto Chaves Junior Sérgio Pinto Chaves Junior Gabriella Lorena Dias Pereira Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Helvécio Cardoso Corrêa Póvoa Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Lívia Azeredo Alves Antunes Leonardo Santos Antunes Leonardo Santos Antunes Leonardo Santos Antunes Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio Natalia Lopes Pontes Póvoa Iorio Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment Frontiers in Oral Health dental pulp necrosis microbiology nucleic acid amplification techniques polymerase chain reaction bacteria |
| title | Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment |
| title_full | Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment |
| title_fullStr | Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment |
| title_full_unstemmed | Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment |
| title_short | Whole-genome amplification as a tool to improve bacterial detection by PCR in microbiological samples after endodontic treatment |
| title_sort | whole genome amplification as a tool to improve bacterial detection by pcr in microbiological samples after endodontic treatment |
| topic | dental pulp necrosis microbiology nucleic acid amplification techniques polymerase chain reaction bacteria |
| url | https://www.frontiersin.org/articles/10.3389/froh.2025.1520945/full |
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