Results: L. aethiopica parasites for the study were obtained from Ethiopia and laboratory analysis was conducted at
The Ohio State University. DNA was extracted from cultured parasites and an internal transcribed spacer located at
the ribosomal region of L. aethiopica genomic DNA was PCR amplified for species identification. Microsatellite
markers were identified using multilocus microsatellite typing. We generated an enriched genomic library, and
using Primer3 software, designed PCR primers to amplify sequences flanking the detected microsatellites.
Subsequent screening of the amplified markers for length variations was performed by gel electrophoresis.
Using a variety of molecular methods, 22 different microsatellite markers were identified and tested for typing
L. aethiopica strains using a number of clinical isolates. Of the 22 markers tested, 5 were polymorphic and showed
distinctive multilocus genotypes, classifying them into four clusters. One marker was found to be specific for
L. aethiopica, discriminating it from other species of Leishmania.