Development and application of an ELISA kit for the detection of milk progesterone in dairy cows

Progesterone (P4) is a steroid gonadal hormone that is mainly produced from the corpus luteum and placenta and has various biological functions, especially reproductive regulation. It is important to establish a specific and sensitive P4 enzyme-linked immunosorbent assay (ELISA) for the study of ovary activity and reproductive disorders. Therefore, we prepared a monoclonal antibody (MAb) in a completed antigen (11α-OH-P4-HS-OVA). Based on the MAb and our previously prepared completed antigen, a highly specific and sensitive ELISA was developed. In the present study, a competitive ELISA for the determination of P4 was described in dairy cow milk. It was found that P4 concentration in milk samples from five pregnant cows was significantly higher than that from five estrus cows. The diagnosis rate for pregnancy and non-pregnancy in 54 dairy cows were 93.3% and 95.8%, respectively, at 19 to 23 days after pregnancy by detecting milk P4 concentration. In summary, the developed ELISA is a potential tool for P4 research and offers an alternative, simple, rapid technique for detecting P4, especially in future large clinical investigations on pregnancy identification and reproductive disorders in dairy farms in China.