The present study describes the use of electrochemiluminescenceimmunoassay (ECL) with the specific human kit for measuring plasma testosterone and progesterone in ovine. This study wascarried out on 16 mixed age animals of the local breed. They were divided into two groups of both sexes, namely Male-1 Group,Male-2 Group, Female-1 Group and Female-2 Group. Blood samples were collected from the jugular vein into tubes containing EDTAand centrifuged at 1500 rpm for 20 min. Plasma was rapidly separated and stored at -20° C until assayed. The reproducibilityinter- and intra-assay of the P4- and T-ECL is satisfactory (2.11-7.3 %). The accuracy (93-102 %) and the test of parallelism were largely acceptable. No cross-reaction was observed with the different hormones including PMSG,hCG, progesterone,testosterone, oxytocin and PGF2α when concentrations of 10 UI/ml and 10-3 UI/ml were used. T concentrations were higher in Male-1 Group than in Male2 Group (1.67 ± 1.15 and 0.38 ± 0.45 ng/ml, respectively). The female-1 Group showed very high P4 concentrations (15.17 ± 4.91 ng/ ml) compared to the Female-2 Group (0.08 ± 0.04 ng/ml). The results obtained clearly show that human ECL system can be used to measure progesterone and testosterone in plasma ovine