Twenty-six isolates of Leishmania parasites of Peruvian origin were studied by isoenzyme electrophoresis of four marker enzymes (ASAT, ALAT, G6PD and GPI), kinetoplast DNA hybridization and monoclonal antibody binding and compared with marker strains of the New World organisms L. b. braziliensis, L. b. guyanensis, L. m. mexicana and L. m. amazonensis. 12 of the isolates studied were of Andean origin; 11 of these were isolated from patients with Andean cutaneous leishmaniasis. The organisms originating from the Peruvian Amazonian forest were isolated from patients with cutaneous (12 cases) or mucocutaneous (2 cases) leishmaniasis. One of the Andean isolates was obtained from an infected phlebotomine vector. 25 of the new isolates were identified as L. braziliensis ssp. according to the three techniques employed. The results of monoclonal antibody binding showed that 23 of the isolates were indistinguishable from L. b. braziliensis. Two isolates identified as L. braziliensis ssp. according to their isoenzyme profiles and k-DNA hybridization patterns could not be classified at the subspecies level. The isolate obtained from the phlebotomine vector could not be identified. No evidence of the existence of parasites of the L. mexicana complex in Peruvian territory was found in this study. The results obtained show a remarkable similarity between Leishmania of Andean origin and L. b. braziliensis. © 1987.