Genetic diversity of Leishmania amazonensis strains isolated in northeastern Brazil as revealed by DNA sequencing, PCR-based analyses and molecular karyotyping

Abstract\ud \ud \ud \ud Background\ud \ud Leishmania (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.\ud \ud \ud \ud Results\ud \ud ITS 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.\ud \ud \ud \ud Conclusion\ud \ud These 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.We thank Lucile FloeterWinter for critical reading of the manuscript and Artur T.L. de Queiroz for initial help with phylogenetic analysis. This work is supported by grants from CNPq, FAPESB and PAPES/FIOCRUZ. J.P.C. de Oliveira was supported by a CNPq fellowship; C.I.O. and F.M.C.F were supported by a FAPESB fellowship. AAC, AB, and CIO are senior investigators from CNPq. AB is a senior investigator for Instituto de Investigação em Imunologia (iii).We thank Lucile Floeter-Winter for critical reading of the manuscript and Artur T.L. de Queiroz for initial help with phylogenetic analysis. This work is supported by grants from CNPq, FAPESB and PAPES/FIOCRUZ. J.P.C. de Oliveira was supported by a CNPq fellowship; C.I.O. and F.M.C.F were supported by a FAPESB fellowship. AAC, AB, and CIO are senior investigators from CNPq. AB is a senior investigator for Instituto de Investigação em Imunologia (iii)

Phylogenetic relationships among strains inferred by Maximum likelihood analysis of the sequences of the ITS1, ITS2 and 5

<p><b>Copyright information:</b></p><p>Taken from "Genetic diversity of strains isolated in northeastern Brazil as revealed by DNA sequencing, PCR-based analyses and molecular karyotyping"</p><p>http://www.kinetoplastids.com/content/6/1/5</p><p>Kinetoplastid Biology and Disease 2007;6():5-5.</p><p>Published online 21 Jun 2007</p><p>PMCID:PMC1919383.</p><p></p>8 S region. The numbers indicate the percentages with which a given branch is supported in 1000 bootstraps replications. The tree is rooted using as outgroup

UPGMA dendrogram based on the combined RAPD and SSR-PCR amplification patterns obtained for strains

<p><b>Copyright information:</b></p><p>Taken from "Genetic diversity of strains isolated in northeastern Brazil as revealed by DNA sequencing, PCR-based analyses and molecular karyotyping"</p><p>http://www.kinetoplastids.com/content/6/1/5</p><p>Kinetoplastid Biology and Disease 2007;6():5-5.</p><p>Published online 21 Jun 2007</p><p>PMCID:PMC1919383.</p><p></p> Bootstrap values above 50% are indicated