In this study, presence of Brucella spp. in Trakya district was investigated. For this purpose, milk samples of 75 unvaccineted cattle raised in various districts, villages and farms, were randomly collected. The collected samples were used for bacterial isolation and detection of Brucella specific DNA. For isolation, the milk samples were inoculated onto Brucella selective media. PCR technique was applied for determination of the specific DNA in the milk samples. As a result of bacteriological studies, Brucella spp. were isolated from three (4%) of total 75 milk samples. One of the isolates was identified as B. melitensis, while the others were B. abortus. Brucella specifical DNA was amplified in 17 (22.66 %) of 75 samples by PCR. The culture positive milk samples were also found to be positive by PCR.