Development of a direct and specific enzymeimmunoassay for the measurement of oestrone sulfate in bovine milk.
A 12-step procedure is described for the synthesis of an oestrone-3-sulfate-6-hemisuccinate-BSA immunogen with oestradiol as starting material and the production of specific polyclonal antibodies. A competitive inhibition-type enzymeimmunoassay has been developed based on these specific antibodies using 3-hemisuccinate-oestrone-peroxidase as conjugate for direct measurement of the hormone in body fluids. The method has a minimum sensitivity of 0.03 ng ml-1 in bovine milk, and satisfactory specificity, recovery and reproducibility. In a small field trial with a group of 20 pregnant cows that were followed throughout gestation, it was shown that the assay is potentially an accurate pregnancy test for assessing the viability of the fetoplacental unit at approximately 100 days after insemination. The assay is well suited for routine testing, particularly as a confirmatory bovine pregnancy test.
Department of Immunology and Diagnostics, Institute of Molecular Biology and Biotechnology, Heraklion, Crete, Greece.
G
G
Hatzidakis
K
K
Katrakili
E
E
Krambovitis
D000818:Animals; D000906:Antibodies; D002417:Cattle; D004966:Estrogens, Conjugated (USP) / Q000032:analysis* / Q000276:immunology; D004970:Estrone / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D005260:Female; D007124:Immunoenzyme Techniques; D008892:Milk / Q000737:chemistry*
D000906:Antibodies; D004966:Estrogens, Conjugated (USP); D004970:Estrone; C017296:estrone sulfate