Amplification of ribosomal internal transcribed spacer 1 (ITS1) sequences from Brazilian clinical isolates of Trypanosomatidae sp.

Promastigote cultures were stablished from two clinical isolates obtained from a patient diagnosed with visceral leishmaniasis in northeast region of Brazil. Clonal colonies were obtained by plating promastigote cultures from each strain. In order to type parasite species, we performed DNA extraction from promastigotes to amplify ribosomal internal transcribed spacer 1 (ITS1) sequences by PCR using primers LITSR and L5.8S as performed previously by Schönian et al. (Diagn Microbiol Infect Dis. 2003 Sep 1;47(1):349–58) . Amplicons were sequenced by Sanger method and multiple sequence alignments showed that the parasites presented higher nucleotide identity with Crithidia species (one-host parasites exclusive for insects) than Leishmania species.