Detection and Identification of Human PathogenicLeishmaniaandTrypanosomaSpecies by Hybridization of PCR-Amplified Mini-exon Repeats

Abstract

A single pair of PCR primers within a conserved region of the mini-exon repeat was used to amplify the repeats from 10 species of pathogenicLeishmaniabelonging to four major clinical groups and also from three species ofTrypanosoma.Oligonucleotide hybridization probes for the detection and identification of the PCR-amplified repeats were constructed from alignments of mini-exon intron and intergenic sequences. The probes generated from mini-exon intergenic regions of theL.(V.)braziliensis, L.(L.)donovani,andL.(L.)mexicanaspecies hybridized specifically to their cognate groups without discriminating between the species within the groups. The probes forL.(L.)majorandL.(L.)aethiopicawere species-specific, while theL.(L.)tropicaprobe also hybridized with theL.(L.)aethiopicamini-exon repeat. The mini-exon intron-derived probes forT. cruzi, T. rangeli,andT. bruceiwere species-specific. This method involving the detection of specific PCR-amplified products produced using a single primer set represents a novel sensitive and specific assay for multiple trypanosomatid species and groups.