Introduction: In the Bel?m Metropolitan Region (BMR), Par? State, Brazil, American cutaneous leishmaniasis (ACL) is endemic; however,
very little is known regarding its causative agents. Therefore, we used our standard diagnostic approach combined with an RNA polymerase
II largest subunit (RNAPOIILS)-polymerase chain reaction (PCR) followed by analysis of restriction fragment length polymorphism
(PCR-RFLP) to identify Leishmania spp. ACL agents in this region. Methods: Thirty-two Leishmania spp. isolates from patients with ACL
in the BMR during 1995-2018 were analyzed. Leishmania spp. DNA samples were amplified using the primers RPOR2/RPOF2, and the
615-bp PCR products were subjected to enzymatic digestion using TspRI and HgaI endonucleases. Results: ACL etiological agents in the
BMR comprised Leishmania (Viannia) lindenbergi (43.7%) followed by Leishmania (Viannia) lainsoni (34.4%), Leishmania (Leishmania)
amazonensis (12.5%), and Leishmania (Viannia) braziliensis (9.4%). Conclusions: To our knowledge, the results of the study revealed for
the first time that L. (V.) lindenbergi and L. (V.) lainsoni are the main ACL agents in BMR.
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