Amplification of small subunit rRNA sequences from promastigote clones of Brazilian clinical isolates of Trypanosomatidae spp.

Promastigote cultures were stablished from two clinical isolates (LHV60 and LHV60a) obtained from a patient diagnosed with visceral leishmaniasis in northeast region of Brazil. Another clinical isolate (HUUFS14) also was used as laboratory reference strain of Leishmania infantum. Clonal colonies were obtained by plating promastigote cultures from each strain. In order to type parasite species, we performed DNA extraction from promastigote culture to amplify small subunit RNA sequences by PCR using primers described by Noyes et al (International Journal for Parasitology v. 29, p. 331-339, 1999). Amplicons were sequenced by Sanger method and phylogenetic analysis showed that the LHV60 and LVH60a parasites were more closely related to Crithidia fasciculata (one-host parasites exclusive for insects), rather than Leishmania species.