Objective:To determine the sand flies species responsible for most transmission of Leishmania major(L major) to human,as well as to determine the main reservoir hosts of the disease. Methods:Sand flies were collected using sticky papers and mounted in Puri’s medium for species identification.Rodents were trapped by live Sherman traps.Both sand flies and rodents were subjected to molecular methods for detection of leishmanial parasite.Results:Phlebotomus papatasi(P.papatasi) was the common species in outdoor and indoor resting places.Employing PCR technique only three specimens of 150 P.papatasi(2%) were found naturally infected by parasites with a band of 350 bp which is equal to the L major parasite.Forty six rodents were captured by Sherman traps and identified.Microscopic investigation on blood smear of the animals for amastigote parasites revealed 1(3.22%) infected Meriones libycus(M.libycus). Infection of this animal to L.major was confirmed by PCR against rDNA loci of the parasite. Conclusions:This is the first molecular report of parasite infection of both vector(P.papatasi) and reservoir(M.libycus) to L major in the region.The results indicated that P.papatasi was the primary vector of the disease and circulating the parasite between human and reservoirs and M. libycus was the most important host reservoir for maintenance of the parasite source in the area. Methods: Sand flies were collected using sticky papers and mounted in Puri’s medium for species identification. Rodents were trapped by live Sherman traps. Both sand flies and rodents were subjected to molecular methods for detection of leishmanial parasite. Results: Phlebotomus papatasi (P. papatasi) was the common species in outdoor and indoor resting places. ImpMENT PCR technique only three specimens of 150 P. papatasi (2%) were found naturally infected by parasites with a band of 350 bp which is equal to the L major parasite. Forty six rodents were captured by Sherman traps and identified. Microscopic investigation on blood smear of the animals for amastigote parasites revealed 1 (3.22%) infected Meriones libycus (M. libycus). Infection of this animal to L. major was confirmed by PCR against rDNA loci of the parasit Conclusions: This is the first molecular report of parasite infection of both vector (P.papatasi) and reservoir (M.libycus) to L major in the region. The results indicated that P.papatasi was the primary vector of the disease and circulating the parasite between human and reservoirs and M. libycus was the most important host reservoir for maintenance of the parasite source in the area.