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Brucellosis is an infectious disease, has a considerable impact on animal health as well as socioeconomic impacts. It causes significant reproductive losses in sexually mature animals. The present study reports the isolation, identification and molecular detection of Brucella abortus. A total of 192 buffalo clinical samples were processed by cultural isolation on BBL Brucella agar plate with selective antibiotic supplements and direct PCR. Out of 192 clinical samples, 7 samples growth yielded on BBL Brucella agar plate and identify Brucella organism by colony character, Gram’s staining, MZN staining, Catalase, KOH and Triple Sugar Iron Agar (TSI) test. | Isolation identification and molecular detection of Brucella abortus from buffaloes in Gujarat India