Identification of leishmania species causing cutaneous leishmaniasis by RAPD-PCR

ABSTRACT

Leishmaniasis, especially cutaneous leishmaniasis is considered as an important health problem in many parts of Iran such as Neishabour, in Razavi Khorasan Province. Since DNA of every parasite such as every organism is specific, this facilitates extensive use of DNA for diagnosis and identification of parasite species. Among these methods, RAPD-PCR seems to be a useful method. Since Neishabour is an important focus for cutaneous leishmaniasis and so far molecular researches haven't been done, this study was done to identify different species of leishmania parasites causing cutaneous leishmaniasis by RAPD-PCR. In this study a total of fifty-seven patients, whom their disease confirmed by direct smear, were recruited and samples isolated and cultured in NNN medium, followed by sub-culturing in RPMI-1640. This study was approved by the local ethics. Then DNA was extracted by proteinase k and desirable samples were amplified by RAPD-PCR method, using four oligoprimers. Electrophoresis patterns from each isolate were compared with the reference strains of L. major, L. tropica and the marker. The results of this study indicated that L. tropica is the responsible parasite for causing cutaneous leishmaniasis, in Neishabour, Northeast of Iran. It seems that Leishmania tropica is the only causative agent of Cutaneous Leishmaniasis in this study area. RAPD-PCR technique is a suitable tools for Leishmania characterization the in epidemiological studies