Abstract
BackgroundLeishmania (Leishmania) amazonensis infection in man results in a clinical spectrum of disease manifestations ranging from cutaneous to mucosal or visceral involvement. In the present study, we have investigated the genetic variability of 18 L. amazonensis strains isolated in northeastern Brazil from patients with different clinical manifestations of leishmaniasis. Parasite DNA was analyzed by sequencing of the ITS flanking the 5.8 S subunit of the ribosomal RNA genes, by RAPD and SSR-PCR and by PFGE followed by hybridization with gene-specific probes.ResultsITS sequencing and PCR-based methods revealed genetic heterogeneity among the L. amazonensis isolates examined and molecular karyotyping also showed variation in the chromosome size of different isolates. Unrooted genetic trees separated strains into different groups.ConclusionThese results indicate that L. amazonensis strains isolated from leishmaniasis patients from northeastern Brazil are genetically diverse, however, no correlation between genetic polymorphism and phenotype were found.
Highlights
Leishmania (Leishmania) amazonensis infection in man results in a clinical spectrum of disease manifestations ranging from cutaneous to mucosal or visceral involvement
L. amazonensis has been isolated from patients with visceral leishmaniasis (VL) and from patients with muco-cutaneous leishmaniasis (ML), in Brazil, these two clinical manifestation are most generally associated with infection by L. chagasi and L. braziliensis [4], respectively
Results rRNA sequencing and analysis To begin examining diversity at the interspecific level, the DNA segment containing internal transcribed spacers (ITS) 1 and 2, as well as the 5.8 S RNA gene, was amplified by PCR using as template DNA from 17 L. amazonensis strains isolated from patients with different clinical manifestations of leishmaniasis, DNA from a reference L. amazonensis strain (M2269) and DNA from a L. major strain (MHOM/81/IL/Friedlin), which was used as an out-group (Table 1)
Summary
Leishmania (Leishmania) amazonensis infection in man results in a clinical spectrum of disease manifestations ranging from cutaneous to mucosal or visceral involvement. We have investigated the genetic variability of 18 L. amazonensis strains isolated in northeastern Brazil from patients with different clinical manifestations of leishmaniasis. Leishmania is an intracellular protozoan parasite that infects humans and causes a wide spectrum of diseases known as leishmaniases. In the American continent, leishmaniasis is caused by a different number of species and infection with L. amazonensis, in particular, produces a wide spectrum of clinical diseases [1]. L. amazonensis has been isolated from patients with visceral leishmaniasis (VL) and from patients with muco-cutaneous leishmaniasis (ML), in Brazil, these two clinical manifestation are most generally associated with infection by L. chagasi and L. braziliensis [4], respectively
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