Abstract

Genotyping Cutaneous Leishmaniasis (CL) species is important for selecting the appropriate method to control and prevent disease, determine the health effects of drugs, prepare and evaluate the vaccine and finally to determine disease vectors and reservoirs. In order to identify Leishmania species, endemic foci of Iran center (Isfahan) was used in this study using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) method. For this purpose, 305 clinical samples were collected from patients referred to health centers in the city of Isfahan. Two samples were collected from each patient for direct slide and culture. Nucleic acid of samples was separated using deoxyribonucleic acid (DNA) extraction kit and kept in 20°C until the test time. Amplify of ITS1 area using L5.8s and LITSr primers showed 350 bp in a number of isolated cases and 450 bp in some others. Enzyme digestion of bands was compared using the enzyme HaeIII with the digested areas of reference strains. The resulted product of a number of isolates was analyzed after determining the sequence. The results showed that 193 isolated cases had 350 bp bonds and after the enzyme digestion revealed 220 and 140 bp bands that were related to the Leishmania major and 7 isolated cases with 350 bp bands that revealed bands 200 and 60 bp after enzymatic digestion that was related toLeishmania tropica and 55 isolated cased had 450 bp bond and after enzyme digestion, revealed 300 and 150 bp parts and after determining the sequence with Crithidia fasciculata and Crithidia luciliae they were matched to about 96%. It seems that by planning new studies and completing current studies, more conclusive results can be accessed and therefore it seems that control, prevention and treatment of the disease in the future may be associated with changed strategies.

Key words: Leishmania, Cutaneous Leishmaniasis, ITS1, polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP).