A direct enzyme immunoassay for 18-hydroxycortisol in urine: a new tool for screening primary aldosteronism.
A microplate enzyme immunoassay has been developed for the measurement of 18-hydroxycortisol in urine. An antiserum was produced by immunization of rabbits with a 3-O-(carboxymethyl)oximino-18-hydroxycortisol-bovine serum albumin conjugate. IgG was isolated from the antiserum and was biotinylated. Newly synthesized p-nitrophenyl ester of the oxime was used for the preparation of steroid-horseradish peroxidase conjugate. After an incubation of diluted urine samples (or standards) and the steroid-enzyme conjugate with the biotinylated antibody, the resulting antigen-antibody complex was separated by adding a portion of the reaction mixture into the avidin-coated microtiter plate. Peroxidase bound to solid phase was measured colorimetrically. The standard curve was linear from 0.25 to 10 ng/well. Intra- and interassay coefficients of variation were 5.5-8.8 and 7.8-8.2%, respectively. The assay was specific except for 18-hydroxycortisone with minor cross reaction. Urinary 18-hydroxycortisol excretion ranged 836-7460 and 26-696 nmol/24 h, respectively, in patients with primary aldosteronism (n = 8) and in control subjects (n = 40). This simple and rapid (< 4 h) assay is suitable for screening patients with primary aldosteronism.
Chiba
H
H
Department of Laboratory Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
Ikegawa
S
S
Kurosawa
T
T
Yoshimura
T
T
Ito
Y
Y
Matsuno
K
K
Kobayashi
K
K
Tohma
M
M
D000818:Animals; D000918:Antibody Specificity; D006801:Humans; D006854:Hydrocortisone / Q000031:analogs & derivatives* / Q000652:urine; D006929:Hyperaldosteronism / Q000175:diagnosis*; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000302:isolation & purification; D011237:Predictive Value of Tests; D011817:Rabbits
D007074:Immunoglobulin G; C033689:18-hydroxycortisol; D006854:Hydrocortisone