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/sup 125/I-radioimmunoassay for unconjugated estroil in serum of pregnant women

Journal Article · · Clin. Chem.; (United States)
OSTI ID:7053622

An /sup 125/I-radioimmunoassay is described which measures unconjugated estriol in the serum of pregnant women. Estriol is extracted into ethyl acetate/hexane, an aliquot is evaporated, and the residue is redissolved in phosphate buffer. The sample is incubated with tracer and antibody at 37/sup 0/C for 15 min and then at 4/sup 0/C for 1 h and 45 min. The antibody-bound fraction is then precipitated with polyethylene glycol and isolated by centrifugation. Because the antigen-antibody complex is stable in the presence of polyethylene glycol, the separation steps are not influenced by timing. Extraction recovery of /sup 3/H-labeled estriol added to a pool of sera from pregnant women averaged 96.7% (SD = 1.3, n = 10). Estriol-supplemented (5, 10, and 20 ..mu..g/liter) serum from men, carried through the entire procedure, showed analytical recovery ranging from 94 to 106%. Structurally analogous steroids normally present in serum of pregnant women exhibit negligible cross reactivity. Day-to-day precision (CV) is 13.3% (3.1 ..mu..g/liter), 6.4% (7.6 ..mu..g/liter), and 5.6% (21.4 ..mu..g/liter) for n = 21. The current reagent cost (about 17 cents per tube), and a total procedural time, including counting, of 5.5 h make this an acceptable assay for routine use.

Research Organization:
Swedish Hospital Medical Center, Seattle
OSTI ID:
7053622
Journal Information:
Clin. Chem.; (United States), Journal Name: Clin. Chem.; (United States) Vol. 24:7; ISSN CLCHA
Country of Publication:
United States
Language:
English