Introduction. The Colombian distribution map of Leishmania species has not been updatedsince seven years ago.Objective. To describe the distribution of Leishmania species isolated from patients attendedat the National Institute of Dermatology during the period 1995 to 2005.Materials and methods. A descriptive study of the geographic distribution of Leishmania specieswas made from 137 isolates obtained from patients consulting the National Institute ofDermatology “Federico Lleras Acosta E.S.E”, with confirmed diagnosis of leishmaniasis. Theclinical history was reviewed to obtain information on the clinical presentation, most probableplace of infection, age and gender. Samples were taken and cultured and the isolates weretyped by monoclonal antibodies, comparing 10% of the results with those obtained by PCR andisoenzymatic patterns.Results. L. panamensis accounted for 74.45% of the 137 isolates studied, L. braziliensis for15.33%, L. guyanensis for 0.73%; L. mexicana complex for 3.65%, L. mexicana 5.11% and the remaining 0.73% corresponded to an isolate which could not be characterized by monoclonalantibodies. The distribution of L. braziliensis, L. panamensis and L. guyanensis was similar tothat reported in previous studies but species of the L. mexicana complex, were found in patientsfrom the departments of Caldas, Santander, Cundinamarca, Caquetá, Casanare, Cauca andValle del Cauca, where they had not been previously reported.Conclusion. Species of the L. mexicana complex display a wider distribution than previouslyreported. The usefulness of species-specific monoclonal antibodies, isoenzymatic patterns,and PCR for identification of L. mexicana and L. amazonensis was limited.