Evaluation of Mulecular Epidemiology of Cutaneous Leishmaniasis In Sabzevar

Document Type : Research Paper

Authors

1 Associate Professor, Parasitology & Medical Mycology Department, Mashhad University of Medical Sciences, Mashhad, Iran

2 Assistant Professor, Iranian Academic Center of Education, Culture& Research, Mashhad Branch

3 Assistant Professor, Virology Mashhad University of Medical Sciences, Mashhad, Iran

4 Specialist In Pediatric Medicine

5 Associate Professor of Social Medicine,University of Medical Sciences, Mashhad, Iran

6 Master Of Science In Parasitology

7 Research Instructor, Iranian Academic Center of Education, Culture& Research, Mashhad Branch

Abstract

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 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
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
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 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.

Keywords


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