Brucellosis is a notable disease, most widespread bacterial zoonoses that still of veterinarian, public health and economic concern in many developing countries including India. Early detection and segregation of infected animals are important in order to control the disease. The present study was done with the objective of isolation and identification of B. melitensis from naturally infected sheep in Sharanpur District, UP (India) using bacteriological and the classical serological techniques; RBPT, STAT, Modified-STAT and i-ELISA. Out of 145 ovine serum samples collected, 48(33.1%) were found to be positive by RBPT taken as a primary screening test. From 48 positive serum samples, 39(81.25%) were found to be positive by STAT, 37(77.08%) by Modified-STAT and 35(72.92%) by i-ELISA. Thus, i-ELISA showed 4 (8.33%) less positive, reduced doubtful by 3 (6.25%) and showed 7 (14.58%) more negative as compared to STAT and 2 (4.16%) less positive, reduced doubtful by 1 (2.08%) and showed 3 (6.25%) more negative to modified-STAT. On the basis of colony morphology, staining characters, phenotypic and biochemical characterizations, the organisms from clinically infected sheep knee joint fluid and aborted foetal samples were isolated and identified as B. melitensis. In addition, two isolates of B. abortus were also identified from liver sample of aborted bovine foetus.