The Persian fallow deer ( Dama mesopotamica ) is a rare species in the world. After presumed extinction of this species in the wild, captive breeding has been used in human controlled environment and introduced to the wild. This species has The obtained sequences were used for comparison of Persian fallow deer with sequences of other Cervidae species available from GenBank . A phylogenetic tree was constructed using neighbor joining method with the number of replications for bootstrap test was set to 1000 . The results showed that the Persian fallow deer and European fallow deer are genetically divergent and distinct species. Also the European fallow deer haplotype, were not found among Iranian Persian fallow deer population, based on the maternal region of D- loop used in this study. However more studies using bio-parental genes are needed to support this hypothesis. As Persian fallow deer has been illegally hunted in the wild, a pair of primers was designed in conserved region of D-loop which amplified a fragment of 588 bp. Samples from other suspected species such as jabber ( Gazella bennettii ), wild goat ( Capra aegagrus ), urial ( Ovis orientalis ), cattle and sheep were obtained. DNA was extracted from these samples and used in PCR amplification. The result showed that the designed primers only amplify the Persian fallow deer samples and no other species was amplified using these primers, suggesting that these primers can be used for defining these species where identification is required. Keywords: Persian Fallow Deer, Dama mesopotamica , Genetic diversity, D-loop region, specific primers