Molecular Identification of Leishmania Species Using Samples Obtained from Negative Stained Smears

Abstract

Background: Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers sus­pected to CL by PCR method.Methods: Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, nega­tive Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed.Results: Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples.Conclusion: Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accu­rate. PCR is recommended for clinically suspected lesions with negative result of direct smear.

Keywords

• Cutaneous Leishmaniasis
• Direct Smear
• Leishmania Tropica
• PCR