kDNA and Molecular Typing of Leishmania spp. of Cutaneous Leishmaniasis Patients in Sistan and Baluchestan Province with Low Amount of Parasite

Abstract

<strong>Background:</strong> Cutaneous Leishmaniasis, is endemically observed in different parts of Iran in two forms of anthroponotic and zoonotic. The identification of both species and the type of disease are beneficial for treatment and prevention. Microscopic identification of <em>Leishmania</em> species has not provided promising efficacy. The aim of this study was to determine the <em>Leishmania</em> species that are responsible for cutaneous leishmaniasis in Zahedan/ Iran by using PCR and PCR-RFLP techniques. <strong>Method:</strong> Direct smears were obtained from cutaneous leishmaniasis suspected individuals with low parasitemia in cutaneous lesions referred to Zahedan health centers. Eventually, the DNA was extracted from smears using DNA extraction kit. PCR was used to amplify both <em>Leishmania</em> kinetoplastic DNA (kDNA) and ITS1 locus of ribosomal DNA. Additionally, PCR-RFLP on ITS1 products was conducted to determine parasite species. <strong>Results:</strong> PCR-RFLP test (detecting ITS1 locus) on all positive samples in microscopic analysis led to the identification of <em>Leishmania major</em> in 52 samples (54.7%), and 43 cases were detected to have <em>Leishmania</em> <em>tropica</em> (45.3%). On the other hand, kDNA-PCR results indicated a frequency of 68 (55.7%) for <em>L. major</em> and 54 (44.3%) for <em>L. tropica</em>. <strong>Conclusion:</strong> Due to the high frequency of kDNA in parasitic genome, PCR-kDNA compared to PCR-RFLP shows a higher efficiency and accuracy not only in identifying infection, but also in determining parasite species, especially among the patients with fewer lesions. This study also indicates that both <em>L. tropica</em> and <em>L. major </em>could be found in Zahedan, with a greater tropical leishmaniasis endemicity.