[Evaluation of mulecular epidemiology of cutaneous leishmaniasis in Sabzevar]

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 aren't 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 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 markerThe 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. L.tropica and L.major both are Etiologic agents ofCutaneous 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

Identification of leishmania species causing cutaneous leishmaniasis by RAPD-PCR

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