Leishmania parasites of the three distinct pathogenic forms of Indian origin (Cutaneous, visceral and post Kala-azar dermal leishmaniasis) were assessed with five selective media for their rate of primary isolation. Leishmania donovani strains showed the higher rate of isolation in modified Tobie's medium, and Leishmania tropica in Tobie's with FCS. However, strains of post Kala-azar dermal leishmaniasis (PKDL) showed 100% isolation in Schneider's with FCS medium. But as a whole modified Tobie's medium showed a higher rate of parasitic isolation for all the three pathogenic forms. In cutaneous leishmaniasis, the dry type and early lesion are also considered significant in the successful isolation of parasite. The growth study of L. donovani strain in the five selective media showed the modified Tobie's medium was best for the multiplication of the parasite.