In this study, polymerase chain reaction technique (PCR) used to detect Brucella melitensis in aborted ewes' milk in Nineveh Province, Iraq using species specific primers of Br. melitensis.Fifty aborted ewes were involved. Blood and milk samples were taken from each ewe. Blood samples used to detect Brucella antibodies in serum using Rose-Bengal plate test (RBPT), whereas PCR were applied on the milk samples to detect Br. melitensis using one pairs of specific primers for the strain of Br. melitensis. Results indicate 35 out of 50 samples (70%) positive reaction by RBPT. From these 35 samples: 26 samples (74.3%) were positive by PCR, while 9 samples (25.7%) were negative. Samples that showed negative reaction by RBPT (15 out of 50 "30%") revealed 8 samples (53.3%) positive by PCR and 7 samples (46.7%) were negative. The total number of milk samples that gave positive reaction by PCR (Br. melitensis) were 34 out of 50 (68%) distributed on Nineveh's regions between 50–100%. The study concluded that using an accurate tests, such as species-specific PCR technique, is an important for reflecting an accurate incidence; since PCR revealed that Br. melitensis existed in aborted ewe's milk in a high percentage in Nineveh, Iraq, which has a public health importance.