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Evaluation of the analytical and diagnostic performance of a digital droplet polymerase chain reaction (ddPCR) assay to detect Trypanosoma cruzi DNA in blood samples

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dc.creatorRamírez, Juan David
dc.creatorHerrera, Giovannyspa
dc.creatorGalindo Hernández, Carolinaspa
dc.creatorCruz-Saavedra, Lissaspa
dc.creatorMuñoz, Marinaspa
dc.creatorFlórez, Carolinaspa
dc.creatorButcher, Robertspa
dc.date.accessioned2020-05-25T23:58:12Z
dc.date.available2020-05-25T23:58:12Z
dc.date.created2018spa
dc.description.abstractBackground: The recent development of novel Polymerase Chain Reaction (PCR) technologies that confer theoretical advantages over quantitative PCR has considerable potential in the diagnosis of low load infections, such as Trypanosoma cruzi in the chronic phase of Chagas disease. We evaluated the utility of the digital droplet (dd)PCR platform in the detection of T. cruzi infection. Methodology/Principal findings: We imported a validated qPCR assay targeting the T. cruzi satellite tandem repeat (TcSTR) region to the ddPCR platform. Following optimization, we tested and repeated a standard curve of TcI epimastigotes to characterise the analytical performance of the assay on the ddPCR platform. We compared this to published qPCR performance data, and the performance of the qPCR assay in our own testing. We subsequently tested a panel of 192 previously characterized DNA specimens, extracted from the blood of individuals with and without T. cruzi infection. The assay performed well on the ddPCR platform, showing a limit of detection of 5 copies/?L or 1 parasite/mL. This was higher than the published limit of detection for qPCR, which was 0.46 parasites/mL. The ddPCR platform was not significantly more accurate than qPCR at any concentration tested. However, the clinical sensitivity and specificity of the assay were both 100% with perfect agreement between qPCR and ddPCR positive and negative result calling in clinical specimens. An average of 9,286 copies of TcSTR were detected per parasite. Conclusions/Significance: The use of the ddPCR platform to run this assay was comparable, but not superior in terms of performance, to the qPCR platform. © 2018 Ramírez et al. http://creativecommons.org/licenses/by/4.0/.eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1371/journal.pntd.0007063
dc.identifier.issn19352727
dc.identifier.issn19352735
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/22821
dc.language.isoengspa
dc.publisherPublic Library of Sciencespa
dc.relation.citationIssueNo. 12
dc.relation.citationTitlePLoS Neglected Tropical Diseases
dc.relation.citationVolumeVol. 12
dc.relation.ispartofPLoS Neglected Tropical Diseases, ISSN:19352727, 19352735, Vol.12, No.12 (2018)spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85059538668&doi=10.1371%2fjournal.pntd.0007063&partnerID=40&md5=c2f74fcf24de4d1fc116e61ac4e88245spa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto Completo)spa
dc.source.instnameinstname:Universidad del Rosariospa
dc.source.reponamereponame:Repositorio Institucional EdocURspa
dc.subject.keywordAdultspa
dc.subject.keywordRoutineeng
dc.subject.keywordProtozoaneng
dc.subject.keywordBlood samplingspa
dc.subject.keywordChagas diseasespa
dc.subject.keywordChildspa
dc.subject.keywordControlled studyspa
dc.subject.keywordDiagnostic test accuracy studyspa
dc.subject.keywordDna extractionspa
dc.subject.keywordDroplet digital polymerase chain reactionspa
dc.subject.keywordEnzyme linked immunosorbent assayspa
dc.subject.keywordEpimastigotespa
dc.subject.keywordFeverspa
dc.subject.keywordGene dosagespa
dc.subject.keywordHemagglutination inhibition testspa
dc.subject.keywordHemagglutination testspa
dc.subject.keywordHepatomegalyspa
dc.subject.keywordHumanspa
dc.subject.keywordImmunofluorescence testspa
dc.subject.keywordLife cycle stagespa
dc.subject.keywordLimit of detectionspa
dc.subject.keywordNonhumanspa
dc.subject.keywordParasite loadspa
dc.subject.keywordParasitological parametersspa
dc.subject.keywordPredictive valuespa
dc.subject.keywordReal time polymerase chain reactionspa
dc.subject.keywordReproducibilityspa
dc.subject.keywordSensitivity and specificityspa
dc.subject.keywordSplenomegalyspa
dc.subject.keywordTandem repeatspa
dc.subject.keywordTrypanosoma cruzispa
dc.subject.keywordTrypomastigotespa
dc.subject.keywordBloodspa
dc.subject.keywordChagas diseasespa
dc.subject.keywordClassificationspa
dc.subject.keywordDiagnostic testspa
dc.subject.keywordEvaluation studyspa
dc.subject.keywordGeneticsspa
dc.subject.keywordIsolation and purificationspa
dc.subject.keywordParasitologyspa
dc.subject.keywordProceduresspa
dc.subject.keywordTrypanosoma cruzispa
dc.subject.keywordProtozoal dnaspa
dc.subject.keywordChagas diseasespa
dc.subject.keywordDiagnostic testseng
dc.subject.keywordDnaeng
dc.subject.keywordHumansspa
dc.subject.keywordReal-time polymerase chain reactionspa
dc.subject.keywordSensitivity and specificityspa
dc.subject.keywordTrypanosoma cruzispa
dc.titleEvaluation of the analytical and diagnostic performance of a digital droplet polymerase chain reaction (ddPCR) assay to detect Trypanosoma cruzi DNA in blood samplesspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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