Establishment of an ELISA for measuring bovine pregnancy-associated glycoprotein in serum or milk and its application for early pregnancy detection.

It has been shown in several mammalian species that during pregnancy, trophoblast cells express a range of pregnancy-associated glycoproteins (PAG). The presence of PAG in the maternal serum of cows may serve as an indicator of pregnancy from day 28 after AI onward. The present study addresses (1) conversion of an existing PAG-RIA to a competitive double antibody ELISA using a polyclonal anti-bPAG-IgG and an anti-rabbit-IgG raised in sheep for coating and (2) application of newly established ELISA to test its suitability for pregnancy detection by measuring PAG in serum or milk. The intraassay coefficients of variation (CV) for the PAG-ELISA were 2-14% for serum and 10-12% for milk; the corresponding interassay CVs were 8-22% and 12-22%, respectively. Pregnancy-associated glycoprotein profiles established in milk and serum of 12 pregnant cows showed a characteristic pattern with measurable amounts from approximately day 20 onwards in serum and from day 60 onwards in milk. In a field trial, serum PAG was determined in 397 cows sampled between 20 and 50 days after insemination. The outcome was that, pregnancy could reliably be diagnosed from day 28 onwards in serum and from day 150 onwards in milk. In conclusion, it may be stated that the established ELISA provides an efficient and reliable means of pregnancy diagnosis that will, in our judgement, gain in popularity with cattle breeding. The ELISA proved to be an adequate and efficient way of measuring PAG in maternal serum or milk and will be a useful means of pregnancy detection in cows.