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PCR-RFLP y RAPD para la tipificación de Leishmania neotropical

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dc.contributor.author Montalvo, A. M.
dc.contributor.author Monzote, L.
dc.contributor.author Fraga, J.
dc.contributor.author Montano, I.
dc.contributor.author Muskus, C.
dc.contributor.author Marín, M.
dc.contributor.author De Doncker, S.
dc.contributor.author Vélez, I. D.
dc.contributor.author Dujardin, J. C.
dc.date.accessioned 2009-06-11T11:47:28Z
dc.date.available 2009-06-11T11:47:28Z
dc.date.issued 2008
dc.identifier.issn 0120-4157
dc.identifier.other ITG-P7B
dc.identifier.other ITG-PLA
dc.identifier.other PARAS
dc.identifier.other U-PROTO
dc.identifier.other FTA
dc.identifier.other UPD14
dc.identifier.other ABSTRACT
dc.identifier.uri http://hdl.handle.net/10390/2682
dc.description.abstract INTRODUCTION: The analysis of the PCR-restriction fragment length polymorphism and random amplified polymorphic DNA have been useful tools for Leishmania identification. OBJECTIVES: Molecular procedures were demonstrated for identification and typing of reference strains of New World Leishmania and their applicability was validated for clinical samples. MATERIALS AND METHODS: DNA was extracted from 16 reference strains of Latin American Leishmania as well as from clinical samples of leishmaniasis patients. A sequence coding for cysteine proteinase B was amplified by PCR and subjected to restriction fragment length polymorphism analysis. The enzyme used was Taq1. For eight of the reference strains, the random amplified polymorphic desoxyribonucleic acid technique (RAPD) was applied. Band patterns for Leishmania species differentiation were established each each method. The sample size of the clinical sample was of 5. RESULTS: PCR products of the cysteine proteinase B gene were obtained for L. braziliensis, L. peruviana, L. panamensis and L. guyanensis. For the other species, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, no amplification occurred. The patterns of restriction fragments revealed band patterns in common for L. peruviana, L. guyanensis and L. panamensis, whereas L. braziliensis had a distinctive pattern. When human samples were examined, amplification occurred for all cases, and the profiles corresponded to the common profile of L. peruviana, L. guyanensis and L. panamensis. The RAPD technique demonstrated reproducible and distinctive patterns for each of the 8 reference strains, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, making possible to differentiate all them. The advantages and limitations of each procedure are discussed. CONCLUSIONS: The combination of RFP and RAPD methodologies provide useful tools to identify medical important species of Leishmania by recognizing DNA sequences characteristic of each species. en
dc.language Spanish en
dc.publisher Instituto Nacional de Salud
dc.subject Protozoology en
dc.subject Leishmania en
dc.subject Identification en
dc.subject Species en
dc.subject Differentiation en
dc.subject PCR-RFLP en
dc.subject RAPD en
dc.subject America, Latin en
dc.title PCR-RFLP y RAPD para la tipificación de Leishmania neotropical en
dc.type Article en
dc.citation.issue 4 en
dc.citation.jtitle Biomédica en
dc.citation.volume 28 en
dc.citation.pages 597-606 en
dc.publisher.place Bogotá sp
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/19462565
dc.citation.jabbreviation Biomédica en


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