We evaluated the polymerase chain reaction (PCR) for the diagnosis of endemic American Tegumentary Leishmaniasis in Salta, Argentina. Diverse Leishmania species, coexistence of mycotic and varicose ulcers, and high endemicity T. cruzi, represent diagnostic challenges in the region. We performed a simplified PCR using sensitive, generic primers on samples obtained by a non-invasive method. We ... [Show full abstract] tested different culture types and clinical specimens with other microorganisms that induce leishmaniasis-like lesions. The PCR had a sensitivity and specificity of 100%. Forty-five patients with presumptive leishmaniasis were compared to the PCR, smears, and the Montenegro skin test (MST). In the same population, the PCR had an increased sensitivity, detecting 25 of 45 cases compared with 16 of 45 for smears and had a higher sensitivity in detecting mucocutaneous lesions. Diagnosis by PCR was supported by clinical presentation, positive MST results, compatible epidemiology, and in some cases histopathologic results or isolation of parasites by culture. These findings indicate the convenience of incorporating this PCR into diagnostic strategies for detecting leishmaniasis in northern Argentina.