diagnosis and characterization of leishmania species in giemsa-stained slides by pcr-rflp

background: direct identification of leishmania species in giemsa-stained slides without parasite culturing in the areas where multiple species exist, is very helpful. this study was designed to isolate leishmani spp. from giemsa-stained smears and to characterize them by pcr technique. methods: a total of 48 giemsa-stained slides from confirmed cases of leishmaniasis were examined under a light micro­scope at×1000 and classified based on grading of leishmania parasites. dna from each slide was extracted separately and sub­jected to pcr. the ribosomal internal transcribed spacer 1 (its1) was amplified with specific primers and the pcr prod­ucts were digested with a restriction enzyme (haeiii). results: of the 48 microscopy-positive slides, 43(89.6%) were positive by pcr-rflp and leishmania species were identi­fied. a statistically significant difference was observed between the both methods ( p < 0.05) and also a concordance was found between microscopy and pcr-rflp (k= 0.55). conclusion: pcr-rflp seems to be an effective method to identify leishmania species from giemsa-stained smears which have been collected from both infected humans and animal reservoir hosts in iran.