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

CutaneousLeishmaniasis, is endemicallyobserved 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 identificationofLeishmania species has
not provided promising efficacy. The aim of this studywas to determine the Leishmania species
that are responsible for cutaneous leishmaniasis in Zahedan/ Iran by using PCR and PCR-RFLP
techniques.
Direct smears were obtained from cutaneous leishmaniasis suspected individualswith
low parasitemia in cutaneous lesions referred to Zahedan health centers. Eventually, the DNA
was extracted from smears usingDNA extractionkit.PCRwas used to amplify bothLeishmania
kinetoplastic DNA (kDNA) and ITS1 locus of ribosomal DNA. Additionally, PCR-RFLP on
ITS1 productswas conducted to determine parasite species.
PCR-RFLP test (detecting ITS1 locus) on all positive samples inmicroscopic analysis
led to the identification of Leishmaniamajor in 52 samples (54.7%), and 43 caseswere detected
to have Leishmania tropica (45.3%). On the other hand, kDNA-PCR results indicated a
frequency of 68 (55.7%) for L.major and 54 (44.3%) for L. tropica.
Due to thehigh frequencyof kDNA in parasiticgenome,PCR-kDNAcompared 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 L. tropica and L. major could be found in Zahedan, with a greater tropical
leishmaniasis endemicity.