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Acta Trop ; 112(3): 239-48, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19660430

ABSTRACT

The identification and characterization of Leishmania are relevant to diagnosis, treatment, eco-epidemiology studies, prophylactic measures and control of the disease. Two strains of Leishmania (MHOM/BR/1971/BH49 and MHOM/BR/1971/BH121), isolated from human cutaneous leishmaniasis, were studied using biological and molecular characteristics, in comparison with WHO reference strains. These studies are important because both strains were incorporated in a vaccine against American cutaneous leishmaniasis, and one of these strains has been used to prepare specific and sensitive antigen for serodiagnosis of visceral leishmaniasis in Brazil. Studies were made on the growth rates of promastigotes in Grace's insect medium, infectivity to C57BL/6 and BALB/c mice, electrophoresic mobility patterns of isoenzymes, random amplification of polymorphic DNA (RAPD), simple sequence repeat-anchored PCR amplification (SSR-PCR) and DNA fingerprinting profiles, infectivity to murine macrophages and cellular immune response. Infections of mice and macrophages were significantly different among the strains studied. Attempts to infect mice with culture promastigotes were unsuccessful with BH121, but BH49 infected BALB/c and C57BL/6 mice. Isoenzyme electrophoretic mobility patterns, RAPD and SSR-PCR using DNA amplified by polymerase chain reaction (PCR) with nine arbitrary primers, as well as DNA fingerprinting studies with a biotin-labeled 33.15 fingerprinting probe showed similar profiles to those of the Leishmania major WHO reference strain.


Subject(s)
Leishmania major/classification , Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Animals , Brazil , Cells, Cultured , Cluster Analysis , Culture Media , DNA Fingerprinting , DNA, Protozoan/genetics , Female , Genotype , Humans , Isoenzymes/analysis , Leishmania major/pathogenicity , Leishmania major/physiology , Leishmaniasis, Cutaneous/pathology , Macrophages/parasitology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Protozoan Proteins/analysis , Random Amplified Polymorphic DNA Technique , Severity of Illness Index , Skin/pathology , Virulence
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