Generation of species-specific DNA probes for Leishmania aethiopica

We report here the cloning of kinetoplast DNA (kDNA) sequences from Leishmania aethiopica in order to develop a specific and sensitive method for the identification of the parasite. Analysis of the cloned kDNA sequences showed different taxonomic specificities demonstrating sequence diversity within the kinetoplast DNA. Cloned whole minicircle hybridized with all Old World Leishmania species tested. Some cloned fragments of minicircle kDNA hybridized with Leishmania species causing cutaneous leishmaniasis in the Old World, but not with the viscerotropic species. Two L. aethiopica-specific clones were found. These clones hybridized with all L. aethiopica isolates tested, but did not react with other Leishmania species. The nucleotide sequence of the L. aethiopica-specific R3 clone is presented. Clones hybridizing with only some of the L. aethiopica isolates were also identified, although none of them showed specificity only for isolates causing localized (LCL) or diffuse (DCL) form of cutaneous leishmaniasis in Ethiopia.