To reduce calving interval through an appropriate reproductive management program, the early detection of pregnancy at the 1st
trimester is essential to enable the prompt re-insemination of non-gravid cows. We aimed to re-evaluate the efficacy of barium chloride assay to detect early pregnancy in cross-bred cows. Urine and blood samples were collected from 72 and 65 artificially inseminated cows, respectively. Pregnancy status at 1st (Day: 1-90) and 2nd (Day: 91-180)trimesters were compared using two different methods: a barium chloride assay from urine samples and a progesterone enzyme linked immune sorbent assay (P4-ELISA) from blood samples, where rectal palpation was considered as gold-standard. We found that although the overall (combining both 1st and 2nd trimesters) true positive, negative predictive value (NPV) and accuracy of P4-ELISA was better, the barium chloride assay performed equally well P4-ELISA in terms of true positive percentage of pregnancy diagnosis (70.8% in barium chloride vs. 85.7% in P4-ELISA; P>0.05) at the 1sttrimester. In addition, specificity of pregnancy diagnosis was higher in barium chloride test than P4-ELISA at the 1sttrimester (100% in barium chloride vs. 66.7% in P4-ELISA). Therefore, barium chloride method could be a good choice under farm and field set up of early pregnancy diagnosis at the 1st trimester in cows.