شناسايي مولكولي گونه هاي عامل ليشمانيوز جلدي در شهرستان سبزوار

Evaluation of Mulecular Epidemiology of Cutaneous Leishmaniasis In Sabzevar

Introduction Cutaneous leishmaniasis is a parasitic infection With an important health problem in many parts of Iran such as Sabzevar, in Khorasan Razavi province. Epidemiological and clinical findings arenʹt sufficient for identification of parasites. Because the host sources are different an accurate identification and diagnosis is necessary before treatment. DNA of every parasite such as every organism is specific. This facilitates extensive use of DNA for diagnostic and identification of parasite species. Molecular methods such as PCR seem to be very useful for this reason. We decided to identify different species of leishmania parasites causing Cutaneous leishmaniasis by PCR in Sabzevar Materials and Methods A Total of 86 patients, whom diseases were confirmed by direct smear, were recruited and samples were isolated and cultured in NNN medium, followed by sub-cultured in RPMI-1640. Then DNA was extracted using four DNA extraction methods. Extracted kinetoplastic DNA was amplified by PCR method using two specific primers. Electrophoresis patterns from each isolate were compared with reference strains of L.major, L.tropica and the marker Results Original article Evaluation of Mulecular Epidemiology of Cutaneous Leishmaniasis In Sabzevar Received: Dec1, 2009; Accepted: Apr 6, 2010 The related bands to amplified products were detected on agarose gel in all samples expected of DNA extracted by boiling method. The results of kDNA gene templets in Electrophoresis gel indicated the leishmania parasite species, causing Cutaneous leishmaniasis, in Sabzevar as 32 samples L.tropica and 54 samples L.major. Conclusion L.tropica and L.major both are Etiologic agents of Cutaneous leishmaniasis in Sabzevar and PCR technique is a suitable tool for the leishmania species characterization in epidemiological studies. The phenol-chloroform based methods are as valuable as DNeasy mini kit (QIAGEN) but more cost effective than kit.