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Authordc.contributor.authorBontempi, Iván 
Authordc.contributor.authorBizai, María 
Authordc.contributor.authorOrtiz Zuñiga, Sylvia 
Authordc.contributor.authorManattini, Silvia 
Authordc.contributor.authorFabbro, Diana L. 
Authordc.contributor.authorSolari Illescas, Aldo 
Authordc.contributor.authorDiez, Cristina 
Admission datedc.date.accessioned2016-12-13T20:55:45Z
Available datedc.date.available2016-12-13T20:55:45Z
Publication datedc.date.issued2016
Cita de ítemdc.identifier.citationInfection, Genetics and Evolution 43 (2016) 123–129es_ES
Identifierdc.identifier.other10.1016/j.meegid.2016.05.026
Identifierdc.identifier.urihttps://repositorio.uchile.cl/handle/2250/141864
Abstractdc.description.abstractDifferent DNA markers to genotype Trypanosoma cruzi are now available. However, due to the low quantity of parasites present in biological samples, DNA markers with high copy number like kinetoplast minicircles are needed. The aim of this study was to complete a DNA assay called minicircle lineage specific-PCR (MLS-PCR) previously developed to genotype the T. cruzi DTUs TcV and TcVI, in order to genotype DTUs TcI and TcII and to improve TcVI detection. We screened kinetoplast minicircle hypervariable sequences from cloned PCR products from reference strains belonging to the mentioned DTUs using specific kDNA probes. With the four highly specific sequences selected, we designed primers to be used in the MLS-PCR to directly genotype T. cruzi from biological samples. High specificity and sensitivity were obtained when we evaluated the new approach for TcI, TcII, TcV and TcVI genotyping in twenty two T. cruzi reference strains. Afterward, we compared it with hybridization tests using specific kDNA probes in 32 blood samples from chronic chagasic patients from North Eastern Argentina. With both tests we were able to genotype 94% of the samples and the concordance between them was very good (kappa = 0.855). The most frequent T. cruzi DTUs detected were TcV and TcVI, followed by TcII and much lower TcI. A unique T. cruzi DTU was detected in 18 samples meantime more than one in the remaining; being TcV and TcVI the most frequent association. A high percentage of mixed detections were obtained with both assays and its impact was discussed. (C) 2016 Elsevier B.V. All rights reservedes_ES
Patrocinadordc.description.sponsorshipCAI + D, Universidad Nacional del Litoral 50120110100160 PI 15-86 FONDECYT Chile 1085154es_ES
Lenguagedc.language.isoenes_ES
Publisherdc.publisherElsevieres_ES
Type of licensedc.rightsAttribution-NonCommercial-NoDerivs 3.0 Chile*
Link to Licensedc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/cl/*
Sourcedc.sourceInfection, Genetics and Evolutiones_ES
Keywordsdc.subjectTrypanosoma cruzies_ES
Keywordsdc.subjectGenotypees_ES
Keywordsdc.subjectMLS-PCRes_ES
Keywordsdc.subjectHybridizationes_ES
Keywordsdc.subjectDTUes_ES
Keywordsdc.subjectMixed infectionses_ES
Títulodc.titleSimple methodology to directly genotype Trypanosoma cruzi discrete typing units in single and mixed infections from human blood sampleses_ES
Document typedc.typeArtículo de revista
Catalogueruchile.catalogadorapces_ES
Indexationuchile.indexArtículo de publicación ISIes_ES


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Attribution-NonCommercial-NoDerivs 3.0 Chile
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 3.0 Chile