The various species of Leishmania cannot be distinguished morphologically, but therntreatment of leishmaniasis depends on the specific causative species. Thus accurate andrnrapid diagnosis of the specific species of Leishmania should be available for effectiverntreatment of patients. Identification of Leishmania species in humans, in insect vectorsrnand reservoir hosts should be available before planning for large-scale epidemiologicalrnsurvey, control programs and possible vaccine and/or drug trials.rnTherefore, the aim of this work was to establish isoenzyme technique at AHRI as a goldrnstandard for species identification and introduce PCR-methods for species-specificrndiagnosis of leishmaniasis directly from clinical materials. In this line, cultured parasitesrnfrom 55 localized cutaneous leishmaniasis (LCL) and 3 diffused cutaneous leishmaniasisrn(DCL) cases), diagnosed at the All Africa Leprosy and Tuberculosis Research,rnRehabilitation and Training Center (ALERT) and from Ochollo village were analyzed.rnSpecies typing with isoenzyme electrophoresis and PCR-based techniques showed that, inrnall the cases, L. aethiopica is the aetiologic agent. Moreover, there was no strain differencernbetween those causing LCL and DCL. The comparison of the isoenzyme and the PCRbasedrnspecies typing techniques confirmed that the intergenic transcribed spacer-1rnpolymerase chain reaction-restriction fragment length polymorphism technique alone couldrnbe used for species typing. The sensitivity and specificity of the technique shows itsrnpotential for species specific diagnosis of leishmaniasis from clinical samples avoiding thernneed for culture. Thus, effective species-specific treatment of patients, which is arnrequirement in leishmaniasis, could be effected in a timely manner.rnKey words: Leishmania aethiopica, Isoenzyme, ITS1-PCR-RFLP, ALERT, Ochollo