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dc.contributor.author
Cura, Carolina Inés
dc.contributor.author
Duffy, Tomas
dc.contributor.author
Lucero, Raúl
dc.contributor.author
Bisio, Margarita María Catalina
dc.contributor.author
Péneau, Julie
dc.contributor.author
Jimenez Coello, Matilde
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Calabuig, Eva
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Gimenez, María J.
dc.contributor.author
Valencia Ayala, Edward
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Kjos, Sonia A.
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Santalla, José
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Mahaney, Susan M.
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Cayo, Nelly M.
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Nagel, Claudia
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Barcan, Laura
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Málaga Machaca, Edith S.
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Acosta Viana, Karla Y.
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Brutus, Laurent
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Ocampo, Susana B.
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Aznar, Christine
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Cuba Cuba, Cesar A.
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Gurtler, Ricardo Esteban
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Ramsey, Janine M.
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Ribeiro, Isabela
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Vandeberg, John I.
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Yadon, Zaida E.
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Osuna, Antonio
dc.contributor.author
Schijman, Alejandro Gabriel
dc.date.available
2016-11-02T17:52:08Z
dc.date.issued
2015-05
dc.identifier.citation
Cura, Carolina Inés; Duffy, Tomas; Lucero, Raúl; Bisio, Margarita María Catalina; Péneau, Julie; et al.; Multiplex Real-Time PCR Assay Using TaqMan Probes for the Identification of Trypanosoma cruzi DTUs in Biological and Clinical Samples; Public Library Of Science; Neglected Tropical Diseases; 9; 5; 5-2015; 3765-3765
dc.identifier.issn
1935-2735
dc.identifier.uri
http://hdl.handle.net/11336/7918
dc.description.abstract
Background. Trypanosoma cruzi has been classified into six Discrete Typing Units (DTUs), designated as TcI?TcVI. In order to effectively use this standardized nomenclature, a reproducible genotyping strategy is imperative. Several typing schemes have been developed with variable levels of complexity, selectivity and analytical sensitivity. Most of them can be only applied to cultured stocks. In this context, we aimed to develop a multiplex Real-Time PCR method to identify the six T. cruzi DTUs using TaqMan probes (MTq-PCR).Methods/Principal Findings. The MTq-PCR has been evaluated in 39 cultured stocks and 307 biological samples from vectors, reservoirs and patients from different geographical regions and transmission cycles in comparison with a multi-locus conventional PCR algorithm. The MTq-PCR was inclusive for laboratory stocks and natural isolates and sensitive for direct typing of different biological samples from vectors, reservoirs and patients with acute, congenital infection or Chagas reactivation. The first round SL-IR MTq-PCR detected 1 fg DNA/reaction tube of TcI, TcII and TcIII and 1 pg DNA/reaction tube of TcIV, TcV and TcVI reference strains. The MTq-PCR was able to characterize DTUs in 83% of triatomine and 96% of reservoir samples that had been typed by conventional PCR methods. Regarding clinical samples, 100% of those derived from acute infected patients, 62.5% from congenitally infected children and 50% from patients with clinical reactivation could be genotyped. Sensitivity for direct typing of blood samples from chronic Chagas disease patients (32.8% from asymptomatic and 22.2% from symptomatic patients) and mixed infections was lower than that of the conventional PCR algorithm.Conclusions/Significance. Typing is resolved after a single or a second round of Real-Time PCR, depending on the DTU. This format reduces carryover contamination and is amenable to quantification, automation and kit production.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Public Library Of Science
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/
dc.subject
Real Time Pcr
dc.subject
Molecular Genotyping
dc.subject
Taqman
dc.subject
Trypanosoma Cruzi
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Bioquímica y Biología Molecular
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Multiplex Real-Time PCR Assay Using TaqMan Probes for the Identification of Trypanosoma cruzi DTUs in Biological and Clinical Samples
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2016-02-01T16:40:30Z
dc.journal.volume
9
dc.journal.number
5
dc.journal.pagination
3765-3765
dc.journal.pais
Estados Unidos
dc.journal.ciudad
San Francisco
dc.description.fil
Fil: Cura, Carolina Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones En Ingeniería Genética y Biología Molecular; Argentina
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Fil: Duffy, Tomas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones En Ingeniería Genética y Biología Molecular; Argentina
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Fil: Lucero, Raúl. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina
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Fil: Bisio, Margarita María Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones En Ingeniería Genética y Biología Molecular; Argentina
dc.description.fil
Fil: Péneau, Julie. Laboratoire Hospitalier et Universitaire-CH Andrée Rosemon; Guayana Francesa
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Fil: Jimenez Coello, Matilde. Universidad Autónoma de Yucatán; México
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Fil: Calabuig, Eva. Hospital Politécnico LA FE; España
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Fil: Gimenez, María J.. Hospital Politécnico LA FE; España
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Fil: Valencia Ayala, Edward. Universidad Peruana Cayetano Heredia; Perú
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Fil: Kjos, Sonia A.. University Of Minnesota; Estados Unidos
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Fil: Santalla, José. Instituto Nacional de Laboratorios en Salud; Bolivia
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Fil: Mahaney, Susan M.. Texas Biomedical Research Institute; Estados Unidos
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Fil: Cayo, Nelly M.. Universidad Nacional de Jujuy; Argentina
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Fil: Nagel, Claudia. Fundacion Favaloro; Argentina
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Fil: Barcan, Laura. Hospital Italiano de Buenos Aires; Argentina
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Fil: Málaga Machaca, Edith S.. Universidad Peruana Cayetano Heredia; Perú
dc.description.fil
Fil: Acosta Viana, Karla Y.. Universidad Autónoma de Yucatán; México
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Fil: Brutus, Laurent. University Paris Descartes; Francia
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Fil: Ocampo, Susana B.. Universidad Nacional de Jujuy; Argentina
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Fil: Aznar, Christine. Laboratoire Hospitalier et Universitaire-CH Andrée Rosemon; Guayana Francesa
dc.description.fil
Fil: Cuba Cuba, Cesar A.. Universidade Do Brasilia; Brasil
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Fil: Gurtler, Ricardo Esteban. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina
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Fil: Ramsey, Janine M.. University Paris Descartes; México
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Fil: Ribeiro, Isabela. Drugs and Neglected Diseases Initiative; Suiza
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Fil: Vandeberg, John I.. Texas Biomedical Research Institute; Estados Unidos
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Fil: Yadon, Zaida E.. Organizacion Mundial de la Salud; Argentina
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Fil: Osuna, Antonio. Universidad de Granada; España
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Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones En Ingeniería Genética y Biología Molecular; Argentina
dc.journal.title
Neglected Tropical Diseases
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0003765
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4437652/
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1371/journal.pntd.0003765
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