Identification by Q-PCR of Trypanosoma cruzi lineage and determination of blood meal sources in triatomine gut samples in México

Triatomine vectors were collected on human dwellings in Michoacán México. Blood meal sources were identified by real time polymerase chain reaction (Q-PCR) using DNA extracted from triatomine guts. The assay was performed with one only specific primer set to amplify a fragment of the mitochondrial 12S ribosomal gene from vertebrate species. Also Trypanosoma cruzi parasites were detected in triatomine gut samples by microscopy and the positive infection was tested in mice. In addition T. cruzi discrete taxonomic units (DTUs) were identified by Q-PCR with two sets of primers that amplify the mini-circle region (miniexon) and 18S ribosomal mitochondrial gene. The sequences obtained from 18S ribosomal gene amplifications confirmed the presence of T. cruzi I and II lineages, and provide evidence of the presence of lineage TcIII and TcIV.

Figure optionsDownload as PowerPoint slideHighlights
► Q-PCR was used for detection of blood meal sources and identified by sequencing.
► 4 lineages of T. cruzi were identified, the most abundant were TcI and TcIV.
► The identification of T. cruzi was done with DNA obtained from triatomine gut.
► Q-PCR and PCR was used to distinguish lineages of T. cruzi, confirm by sequence.
► Q-PCR is useful for detection of fresh/dried blood meals from triatomine gut.