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The nanostructure, as a transducer, was utilized to immobilize a <jats:italic>Brucella<\/jats:italic>-specific probe and fabrication of a genosensor and the process of immobilization and hybridization was detected by electrochemical methods, using methylene blue as a redox marker. The proposed method for detection of the complementary sequence, sequences with base-mismatched (one-, two- and three-base mismatches) and the sequence of non-complementary sequence was assayed. The fabricated genosensor was evaluated for the assay of the bacteria in the cultured and human samples without polymerase chain reactions (PCR). 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