Parasitological studies
Skin lesions
SKIN LESIONS, STERILE, LYSIS
In this study, the smears were taken for diagnosis of CL. Skin lesions were cleaned with antiseptic agent (70% alcohol), and then using a sterile lancet, 2–3 mm superficial incisions were made on the edges of the lesions. Two smears of dermal tissue scrapings were prepared from each patient, then air-dried, fixed in methanol, and giemsa-stained. Each smear was completely scanned for Leishman body. Samples were then screened via nested PCR. Using a sterile surgical blade, the dry smear was scraped from slide, and inserted into micro tube containing 200 μl of lysis buffer (50 mg ethylene diamine tetra-acetic acid (EDTA) pH 7.4, 50 mM NaCl, 200 μg proteinase K/ml, and 1% Triton X-100). The mixture was incubated at 56 °C for 3 h. The lysate was extracted with phenol–chloroform, and finally re-suspended in 50 µl double-distilled water and kept at − 20 °C until used. Characterization of the CL isolates was done using nested PCR by specific primers of CSB1XR, and CSB2XF as external primers, and also LiR, and 13Z as internal primers, which can specifically differentiate CL species [