Histopathological Findings
Direct smears from the exudate of fluid of the lupoid lesion were stained with special stains Gram, Erlich-Ziehl-Nielsen (EZN), Periodic Acid-Schiff (PAS), Giemsa for detection of bacteria, mycobacteria, fungi and leishmania amastigotes, respectively. A punch biopsy aspiration fluid from lesions was used, and a part of the exudate of fluid of the lupoid lesion was used for the identification ofLeishmania species by Real Time PCR. Application of real time PCR that targeted the ITS1 region of Leishmania species using the DNAs obtained from the lesion of the patient. In this real time PCR, melting temperature of L. major reference control and sample is 85°C as described previously2.