Brucella spp. are important infectious agents in bovine abortions worldwide. The bacteriological culture of Brucella spp. is fastidious and time consuming procedure as a classical laboratory method. Brucella spp. can be detected by using different molecular techniques. The aim of this study is to develop a PCR technique with an internal control for detection of Brucella spp. from bovine abortion samples. For this purpose, the sensitivities of three different primer pairs (BgF/BgR, B4/B5 and JP-R/JP-F) were compared. Bovine 12S gene specific primer pairs (12SM-FW/12SBT-REV2) were used as an internal control. The sensitivity of BgF/BgR primers was found higher than the other primer sets. A PCR assay was developed by combining BgF/BgR primer sets and primers for bovine 12S. This protocol was tested and validated by using abomasal contents of two Brucella-positive and eighteen Brucella-negative clinical samples. In conclusion, the developed PCR method with an internal positive control has a potential for use in direct detection and identification of the Brucella spp. from bovine abortion samples.