Development of a direct microplate enzyme immunoassay for the determination of pregnanediol-3 alpha-glucuronide in urine.
A sensitive direct enzyme immunoassay for urine pregnanediol-3 alpha-glucuronide was developed. The assay system involves the use of an antiserum against pregnanediol-3 alpha-glucuronide and an enzyme-labelled antigen chemically prepared by linking beta-D-galactosidase to 20 alpha-hydroxy-5 beta-pregnane 3(O-carboxymethyl)oxime. Free from antibody-bound antigen was separated by a solid-phase double antibody method, using a microplate coupled with goat anti-rabbit gamma-globulin. This solid-phase enzyme immunoassay for urine pregnanediol-3 alpha-glucuronide was validated in terms of specificity, accuracy and sensitivity. When urine samples were assayed for pregnanediol-3 alpha-glucuronide, the results obtained by the solid phase enzyme immunoassay and conventional radioimmunoassay methods agreed well (n = 30, r = 0.922). This assay system has an advantage over radioimmunoassay, because it does not require the use of radioisotopes. The procedure of this method is very simple, since it does not require purification steps of the biological samples.
Mitsuma
M
M
Department of Obstetrics and Gynecology, Teikyo University School of Medicine, Tokyo, Japan.
Yoshimura
S
S
Kambegawa
A
A
Okinaga
S
S
Arai
K
K
D005260:Female; D005498:Follicular Phase; D006801:Humans; D007124:Immunoenzyme Techniques; D008183:Luteal Phase; D011247:Pregnancy; D011276:Pregnanediol / Q000031:analogs & derivatives* / Q000652:urine; D011786:Quality Control; D011863:Radioimmunoassay; D012016:Reference Values
C022270:pregnanediol-3 alpha-glucuronide; D011276:Pregnanediol