Background: Cell-mediated immunity (CMI) plays an important role in resistance against leishmaniasis. Leishmanin test (Montenegro skin test) is a method in evaluation of immune system reaction against cutaneous leishmaniasis. The TDR/WHO reference Leishmanin produced by the Pasteur Institute of Iran is used worldwide. Two other antigens produced by Razi Serum Institute of Iran, Killed Leishmania Major (K.L.M) and Autoclaved Leishmania Major (A.L.M), take advantage of vaccine strains of L.major, and under supervision of WHO, have been investigated as leishmaniasis vaccine. Objective: KLM, ALM and standard Leishmanin as skin test wee compared in this study. Patients and Methods: Fifty patients with cutaneous leishmaniasis were randomly selected. They attended the educational clinics of Isfahan University of Medical Sciences. All 3 antigens were injected intradermally in them and the size of erythema and induration were read and recorded after 48 hours. Data were grouped and analyzed with analysis of variance (ANOVA). Results: There was significant difference between mean of erythema and induration sizes of antigens in two by two comparison. The mean of erythema and induration sizes due to K.L.M were greater than A.L.M, and mean of erythema and induration sizes due to A.L.M were greater than Pasteur Institute reference leishmanin (P<0.0001). Conclusion: Although the mean of erythema and induration sizes due to K.L.M were greater than others, use of A.L.M seems to be more practical compared to. K.L.M, because of simple storage method of A.L.M, and more sensitive than Pasteur Institute reference leishmanin.