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Analytical performance of Four Polymerase Chain Reaction (PCR) and real time PCR (qPCR) assays for the detection of six Leishmania species DNA in Colombia

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dc.creatorLeón, Cielo M.spa
dc.creatorMuñoz, Marinaspa
dc.creatorGalindo Hernández, Carolinaspa
dc.creatorAyala, Martha S.spa
dc.creatorFlórez, Carolinaspa
dc.creatorTeherán, Aníbalspa
dc.creatorCubides, Juan R.spa
dc.creatorRamírez, Juan Davidspa
dc.date.accessioned2020-05-26T00:08:34Z
dc.date.available2020-05-26T00:08:34Z
dc.date.created2017spa
dc.description.abstractLeishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America. © 2017 León, Muñoz, Hernández, Ayala, Flórez, Teherán, Cubides and Ramírez.eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.3389/fmicb.2017.01907
dc.identifier.issn1664302X
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/24094
dc.language.isoengspa
dc.publisherFrontiers Media S.A.spa
dc.relation.citationIssueNo. OCT
dc.relation.citationTitleFrontiers in Microbiology
dc.relation.citationVolumeVol. 8
dc.relation.ispartofFrontiers in Microbiology, ISSN:1664302X, Vol.8, No.OCT (2017)spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85030648499&doi=10.3389%2ffmicb.2017.01907&partnerID=40&md5=01d94b9c9607092cb0dfb85f59c8f628spa
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.keywordHeat shock protein 70spa
dc.subject.keywordKinetoplast dnaspa
dc.subject.keywordMolecular markerspa
dc.subject.keywordRna 18sspa
dc.subject.keywordAnalytical parametersspa
dc.subject.keywordAnticipated reportable rangespa
dc.subject.keywordArticlespa
dc.subject.keywordColombiaspa
dc.subject.keywordControlled studyspa
dc.subject.keywordDna extractionspa
dc.subject.keywordGene amplificationspa
dc.subject.keywordLeishmaniaspa
dc.subject.keywordLimit of detectionspa
dc.subject.keywordMeasurement accuracyspa
dc.subject.keywordMicroorganism detectionspa
dc.subject.keywordNonhumanspa
dc.subject.keywordPolymerase chain reactionspa
dc.subject.keywordPost hoc analysisspa
dc.subject.keywordQuantitative analysisspa
dc.subject.keywordReal time polymerase chain reactionspa
dc.subject.keywordReproducibilityspa
dc.subject.keywordSensitivity and specificityspa
dc.subject.keywordAnalytical performancespa
dc.subject.keywordLeishmaniaspa
dc.subject.keywordMolecular diagnosisspa
dc.subject.keywordPcrspa
dc.subject.keywordQpcrspa
dc.titleAnalytical performance of Four Polymerase Chain Reaction (PCR) and real time PCR (qPCR) assays for the detection of six Leishmania species DNA in Colombiaspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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