Amplification of glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) partial sequences from promastigote clones of Brazilian clinical isolates of Trypanosomatidae sp.

Promastigote cultures were stablished from two clinical isolates (LVH60 and LVH60a) 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 promastigotes to amplify glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) sequences by PCR using primers described by Borghesan et al. (Protist, 164:129-152, 2013). Amplicons were sequenced by Sanger method and phylogenetic analysis showed that the LHV60 and LVH60a parasites were more related to Crithidia fasciculata (one-host parasites exclusive for insects), rather than Leishmania species.