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Identification and biochemical characterization of Leishmania strains isolated in Peru, Mexico, and Spain
Authors:Rodríguez-González Isabel  Marín Clotilde  Vargas Franklin  Córdova Ofelia  Barrera Mario  Gutiérrez-Sánchez Ramón  Alunda Jose María  Sánchez-Moreno Manuel
Affiliation:Instituto de Biotecnología, Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, C/Severo Ochoa s/n. 18071 Granada, Spain.
Abstract:Eight Leishmania promastigotes were isolated from different geographical areas: three (LP1, LP2, and LP3) from the provincial department La Libertad and the fourth (LP4) from the department of Cajamarca (northern Peru); another three (LM1, LM2, and LM3) in the province of Campeche (Mexico); and the last (LS1) from a clinical case of a dog in Madrid (Spain). The isolates were characterized by carbohydrate cell-surface residues using agglutinations with four purified lectins, by isoenzyme analysis using different isoenzymes, by analysis of kinetoplast DNA (kDNA) restriction fragment length polymorphism using four different restriction endonucleases and by the final metabolite patterns after in vitro culture. These isolates were compared with four reference strains and typified as: Leishmania (Leishmania) donovani, two strains of L. (L.) infantum, and one species of L. (Viania) peruviana. According to our results and the statistical study, the Peruvian isolates represent three different strains: one would be L. (V.) peruviana, another the strain isolated in Cajamarca (LP4) and the third would include the three strains from the department of La Libertad (LP1, LP2, and LP3), these latter three isolates being phylogenetically closer to the reference strain L. (L.) donovani. Meanwhile, the three isolates from Mexico form a group with close phylogenetic relationships to each other. The isolate from Spain belongs to the species L. (L.) infantum. Thus, a close correlation was drawn between the identity of each strain and its geographical origin.
Keywords:Leishmania species   In vitro culture   Lectin agglutination: Con A, lectin from Canavalia ensiformis   VV, lectin from Vicia villosa   WGA, lectin from Triticum vulgaris   PNA, lectin from Arachis hypogae   Isoenzyme electrophoresis: ME, malic enzyme [EC 1.1.1.40]   MDH, malate dehydrogenase [EC 1.1.1.37]   IDH, isocitrate dehydrogenase [EC 1.1.1.42]   GPI, glucose phosphate isomerase [EC 5.3.1.9]   PGM, phosphoglucomutase [EC 2.7.5.1]   SOD, superoxide dismutase [EC 1.15.1.1]   kDNA restriction pattern: kinetoplast deoxyribonucleic acid   1H NMR, proton nuclear magnetic resonance spectroscopy   MEM, minimal essential medium   EDTA, ethylenediaminetetraacetic acid   NaCl, sodium chloride   DNA, deoxyribonucleic acid
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