Objective: to investigate Leishmania species in a series of autochthonous cutaneous leishmaniasis (CL) cases in Amapá State, Brazilian Amazon. Methods: this was a descriptive ecological study carried out from January-October/2018 at a reference center for CL diagnosis in Amapá; individuals with CL receiving care from January-May/2018 were recruited; clinical data and skin biopsies were obtained; from extracted DNA (phenol-chloroform) we amplified the hsp70-234 gene region (PCR) for nucleotide sequencing (Applied Biosystems: ABI3500XL). Results: 38 individuals were interviewed, examined and diagnosed; men predominated (28/38; mean age=32.5±11.3); lesions (most ulcers: 37/38) measuring 0,4-10mm (34/38) and ≥11mm (4/38) were multiple in 20/38 individuals; diagnosis of L. braziliensis (1), L. naiffi (1), L. infantum (1), L. (Viannia) sp. (1), L. amazonensis (2) and L. guyanensis (32); individuals infected with L. guyanensis (32/38) lived in 9/10 municipalities represented in the sample, and 17/32 of these had multiple lesions. Conclusion: presence of Leishmania guyanensis predominated and was frequently associated with multiple lesions.