Detection of fetal twins in sheep using a radioimmunoassay for pregnancy-specific protein B1.

Ovine pregnancy-specific protein E (oPSPB) was isolated from sheep placentas. Antiserum to oPSPB was developed in rabbits. A quantitative RIA was developed and used to assay the serum concentrations of oPSPB during and after pregnancy in ewes bearing single or twin fetuses. Suffolk and Panama ewes, kept with rams equipped with a marking harness, were checked daily for breeding marks as an indication of estrus and bled daily between 10 and 30 d after marking. Ovine PSP became detectable at 19.7 +/- 0.14 (mean +/- SE) d after breeding and increased steadily to d 30. Panama oPSPB concentration increased at a greater rate than that of Suffolks (breed X day interaction, P <.0.01). Ten ewes were bled twice weekly 3 wk before their expected date of lambing and weekly for 7 wk postpartum. Serum concentrations differed considerably between prepartum ewes, but concentration remained stable within the period of 20 d prepartum. Following parturition, oPSPB concentrations dropped rapidly. In nine ewes, oPSPB was last detectable at 12.78 +/- 2.26 (mean +/- SE) d postpartum. In the 10th ewe, oPSPB was 0.65 ng/mL at the last sample on d 46 postpartum. To determine the effect of fetal number on oPSPB concentrations, ewes in which estrus was synchronized were bled at d 18, 25, 38, 60, 90, and 120 after breeding. Ewes were killed at d 60, 90, 120, and 148 and fetal number determined. There was a significant (P < 0.01) difference in the log of oPSPB concentrations according to number of fetuses, day postbreeding, and the day X fetal number interaction. At 60, 90, and 120 d, the test was 82, 78, and 82% correct in detecting twins and single fetuses. With a combined 60 to 120 d of gestation range, the test was 78% correct at distinguishing twin from single pregnancies. The RIA provided values that were predictive. Ovine PSPB was detectable in sera very early in gestation, was not detectable within a few weeks after parturition, and was unusually higher in ewes bearing twins.