Design and validation of a quantitative polymerase chain reaction test for the identification and quantification of uncultivable bacteria associated with periodontitis

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dc.contributor.authorCastillo, Yormaris
dc.contributor.authorDelgadillo, Nathaly Andrea
dc.contributor.authorNeuta, Yineth
dc.contributor.authorIniesta, Margarita
dc.contributor.authorSanz, Mariano
dc.contributor.authorHerrera, David
dc.contributor.authorPianeta, Roquelina
dc.contributor.authorLafaurie, Gloria Inés
dc.contributor.authorCastillo, Diana Marcela
dc.contributor.orcidCastillo, Yormaris [0000-0002-8673-8449]
dc.contributor.orcidDelgadillo, Nathaly Andrea [0000-0001-8842-549X]
dc.contributor.orcidNeuta, Yineth [0000-0001-9350-0941]
dc.contributor.orcidIniesta, Margarita [0000-0001-7557-4023]
dc.date.accessioned2023-07-19T16:32:13Z
dc.date.available2023-07-19T16:32:13Z
dc.date.issued2023
dc.description.abstractenglishObjective: This study aimed to standardize a quantitative polymerase chain reaction (qPCR)-based test to identify and quantify the uncultivable bacteria associated with periodontitis. Methods: The standardization of qPCR, the curves for the quantification of Eubacterium saphenum, Eubacterium brachy, Desulfobulbus oralis, and Filifactor alocis were developed by cloning the 16 S rRNA target gene fragment, using the GEMTEasy vector. The qPCRs were validated in 55 subgingival biofilm clinical samples, from different stages of periodontitis and from periodontally healthy/gingivitis individuals, which were previously evaluated by next-generation sequencing (NGS). The results obtained by the two methods were compared by the concordance of Cohen's Kappa index, and sensitivity, specificity, receiver operating characteristic (ROC) curve, and predictive values were established. Results: obtained by the two methods were compared using the concordance of Cohen's Kappa index, and sensitivity, specificity, predictive values, and ROC curves were generated. The qPCR test was standardized with efficiencies between 90% and 100% and R2: 0.997–0.999. Concordance between the qPCR and NSG was moderate to F. alocis (agreement 78.2%; kappa 0.56, p < 0.05) and fair to the other microorganisms (agreement 67.27%−72.73; kappa 0.37–0.38, p < 0.05). qPCR exhibited a high sensitivity (82.2–100%) and specificity (100%) for E. brachy, E. saphenum, and F. alocis. Sensitivity was lower to D. oralis. Conversely, qPCR demonstrated higher sensitivity to E. saphenum than NSG (100 vs. 68.1). Conclusions: The uncultivable microorganisms associated with periodontitis, D. oralis, E. brachy, E. saphenum, and F. alocis can be detected and quantified with the newly developed and validates qPCR test.eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1016/j.archoralbio.2023.105758
dc.identifier.instnameinstname:Universidad El Bosquespa
dc.identifier.issn0003-9969
dc.identifier.reponamereponame:Repositorio Institucional Universidad El Bosquespa
dc.identifier.repourlrepourl:https://repositorio.unbosque.edu.co
dc.identifier.urihttps://hdl.handle.net/20.500.12495/11082
dc.language.isoeng
dc.publisherElsevierspa
dc.publisher.journalArchives of Oral Biologyspa
dc.relation.ispartofseriesArchives of Oral Biology, 0003-9969, 154, 2023, 1 - 8spa
dc.relation.urihttps://www.sciencedirect.com/science/article/pii/S0003996923001462?pes=vor
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internacional*
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.rights.accessrightshttps://purl.org/coar/access_right/c_abf2
dc.rights.localAcceso abiertospa
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectEficacia de la PCRspa
dc.subjectQPCRspa
dc.subjectNúmero de copiasspa
dc.subjectBacterias oralesspa
dc.subjectPeriodontitisspa
dc.subject.keywordsPCR efficiencyspa
dc.subject.keywordsQPCRspa
dc.subject.keywordsCopy numberspa
dc.subject.keywordsOral bacteriaspa
dc.subject.keywordsPeriodontitisspa
dc.titleDesign and validation of a quantitative polymerase chain reaction test for the identification and quantification of uncultivable bacteria associated with periodontitisspa
dc.title.translatedDesign and validation of a quantitative polymerase chain reaction test for the identification and quantification of uncultivable bacteria associated with periodontitisspa
dc.type.coarhttps://purl.org/coar/resource_type/c_6501
dc.type.coarversionhttps://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.driverinfo:eu-repo/semantics/article
dc.type.hasversioninfo:eu-repo/semantics/publishedVersion
dc.type.localArtículo de revista

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