Molecular Identification of Leishmania Species Causing Cutaneous Leishmaniasis in Mashhad, Iran

Cutaneous leishmaniasis is considered an important health problem in many parts of Iran, especially in Mashhhad. Various species of leishmania are causing the disease. Identification of leishmania is helpful for the control and prevention of the disease. Althought epidemiological and clinical findings are necessary, they are not sufficient for identification of cutaneous leishmaniasis. Difficulties in identification of these parasites, due to the similar morphology of amastigote and promastigote in different species, have been solved by using molecular techniques such as PCR amplification of kDNA. In order to identify leishmania species causing CL in Mashhad by a definite molecular technique (PCR method), a study was undertaken over a 12 months period (Apr. 2004- Feb. 2005) in department of Parasitology, Ghaem hospital, and Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran. A total of twenty one patients were recruited and samples were isolated and cultured in N.N.N medium followed by sub-cultur in RPMI-1640. Then, DNA was extracted by protein as k and amplified by specific primers of kDNA. The PCR product was analysed by gel electrophoresis using 2% agarose. The gel was stained with ethidium bromide and visible band with the presence of 620 bp fragment indicates of leishmania major and 800 bp indicates of leishmania tropica. The results indicated that 19 of the isolates were identified as L. tropica and the other two were identified as L. major.Discussion & As demonstrated by previous investigations, Mashhad had been known as an endemic focus for anthroponotic cutaneous leishmaniasis (ACL); however, it is now concluded that both ACL and zoonotic cutaneous leishmaniasis (ZCL) are present in Mashhad and L. tropica is the dominant species.