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- IOC - Artigos de Periódicos [12642]
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SCREENING AND CHARACTERIZATION OF RAPD MARKERS IN VISCEROTROPIC LEISHMANIA PARASITES
Autor(es)
Afiliação
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
University of Khartoum. Institute of Endemic Diseases. Khartoum, Sudan.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.
University of Khartoum. Institute of Endemic Diseases. Khartoum, Sudan.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
University of Khartoum. Institute of Endemic Diseases. Khartoum, Sudan.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.
University of Khartoum. Institute of Endemic Diseases. Khartoum, Sudan.
Université Tunis el Manar. Institut Pasteur de Tunis. Laboratoire d’Epide´miologie Mole´culaire et de Pathologie Expe´rimentale (LR11IPT04)/Laboratoire d’Epide´miologie et d’Ecologie Parasitaire (LR00SP04). Tunis, Tunisia.
Resumo em Inglês
Visceral leishmaniasis (VL) is mainly due to the Leishmania donovani complex. VL is endemic in many countries worldwide
including East Africa and the Mediterranean region where the epidemiology is complex. Taxonomy of these pathogens is
under controversy but there is a correlation between their genetic diversity and geographical origin. With steady increase in
genome knowledge, RAPD is still a useful approach to identify and characterize novel DNA markers. Our aim was to identify
and characterize polymorphic DNA markers in VL Leishmania parasites in diverse geographic regions using RAPD in order to
constitute a pool of PCR targets having the potential to differentiate among the VL parasites. 100 different oligonucleotide
decamers having arbitrary DNA sequences were screened for reproducible amplification and a selection of 28 was used to
amplify DNA from 12 L. donovani, L. archibaldi and L. infantum strains having diverse origins. A total of 155 bands were
amplified of which 60.65% appeared polymorphic. 7 out of 28 primers provided monomorphic patterns. Phenetic analysis
allowed clustering the parasites according to their geographical origin. Differentially amplified bands were selected, among
them 22 RAPD products were successfully cloned and sequenced. Bioinformatic analysis allowed mapping of the markers
and sequences and priming sites analysis. This study was complemented with Southern-blot to confirm assignment of
markers to the kDNA. The bioinformatic analysis identified 16 nuclear and 3 minicircle markers. Analysis of these markers
highlighted polymorphisms at RAPD priming sites with mainly 59 end transversions, and presence of inter– and intra–
taxonomic complex sequence and microsatellites variations; a bias in transitions over transversions and indels between the
different sequences compared is observed, which is however less marked between L. infantum and L. donovani. The study
delivers a pool of well-documented polymorphic DNA markers, to develop molecular diagnostics assays to characterize and
differentiate VL causing agents.
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