Measurement of ovine pregnancy-associated glycoprotein (PAG) during early pregnancy in Lacaune sheep

This study describes ovine pregnancy-associated glycoprotein (ovPAG) concentrations in 20 Lacaune sheep during early pregnancy. Measurements were performed by using semi-purified ovPAG as standard, tracer and immunogens for antibody production in rabbits. Antisera R780 (against ovPAG(57+59kDa)) and R805 (against ovPAG5(58+61kDa)) were used respectively in RIA-780 and RIA-805. Blood samples were collected at days 0, 18, 20, 22 and 25 after artificial insemination. From day 18 after breeding onward, the mean ovPAG concentration was significantly higher (p < 0.001) in plasma samples from pregnant ewes (n = 17) than in non-pregnant ones (n = 3). The specific activity of the tracer was 11 760 Ci/mmol in RIA-780 and 14 900 Ci/mmol in RIA-805. The minimal detection limits for RIA-780 and RIA-805 were 0.2 ng/ml and 0.3 ng/ml, respectively. The intra-assay CV of samples with low (1.0 ng/ml), medium (2.5 ng/ml) and high (4.0 ng/ml) PAG concentrations were 3%, 6% and 9% for RIA-780 and 8%, 9% and 5% for RIA-805. The inter-assay CV in the same samples were 13%, 12% and 7% for RIA-780 and 13%, 11% and 5% for RIA-805. The recovery was higher than 95% in both assays. No cross-reaction was observed with members of aspartic proteinase family as well as with other tested proteins. In both RIA-780 and RIA-805, inhibition of the binding of the tracer by antisera was parallel between standard curve and serial dilutions of pregnant ewe samples. In conclusion, the two homologous RIA systems are suitable for early quantification of ovPAG concentrations in ewe plasma samples from day 18 after breeding.