[Enzyme immunoassay of 17-hydroxyprogesterone in dried blood spot on filter paper using specific antibody for 17-hydroxyprogesterone].
Specific antiserum for 17-hydroxyprogesterone (17-OH-P) was prepared by immunizing 7 alpha-(2-carboxyethylthio)-17-OH-P conjugated bovine serum albumin (BSA) in rabbits. Using this antiserum, 17-OH-P enzyme immunoassay for dried blood spots on filter paper was established. As a label, alkaline phosphatase was coupled covalently with 7 alpha-carboxy-methylthio-17-OH-P by carbodiimide method. B/F separation was carried out by the addition of anti-rabbit IgG goat antiserum. All specimens used were punched out with a paper puncher of 3mm diameter. The assay sensitivity was 2pg/tube, which was estimated by two standard deviation at zero concentrations of the calibration curve. Cross reactivities of this antibody were as follows: 11-deoxycortisol (8.21%), 17-OH-pregnenolone (3.33%), progesterone (1.67%), 11-deoxycorticosterone (0.31%), cortisol (0.16%), pregnenolone-3-sulfate Na salt (0.03%), dehydroepiandrosterone (DHEA) (less than 0.03%), 16 alpha-OH-DHEA (less than 0.03%), DHEA-3-glucuronide (less than 0.03%), DHEA-3-sulfate Na salt (less than 0.03%), pregnenolone (less than 0.02%). Intra- and inter-assay coefficient of variations were 4-14% and 9-18%, respectively. In normal babies, 17-OH-P concentrations measured directly (without sample extraction) were below 23pg/disk (n=204). The histogram of 17-OH-P level in normal babies obtained by the direct method was distributed lower than that obtained by the enzyme immunoassay system (range: 4-79pg/disk, n=268) which used antibody raised against 17-OH-P-3-O-carboxymethyloxime conjugated BSA (Enzaplate, Sapporo Diagnostic Laboratory).(ABSTRACT TRUNCATED AT 250 WORDS)
Kobayashi
Y
Y
Clinical Chemistry Laboratory, Kobe Women's College of Pharmacy.
Nishiguchi
Y
Y
Watanabe
F
F
Yamairi
T
T
Hase
Y
Y
Tsuruhara
T
T
Miyai
K
K
D019326:17-alpha-Hydroxyprogesterone; D000818:Animals; D001801:Blood Stains; D003429:Cross Reactions; D006801:Humans; D006908:Hydroxyprogesterones / Q000097:blood* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D007124:Immunoenzyme Techniques; D007231:Infant, Newborn; D007234:Infant, Premature / Q000097:blood; D011237:Predictive Value of Tests; D011817:Rabbits
D006908:Hydroxyprogesterones; D007106:Immune Sera; D019326:17-alpha-Hydroxyprogesterone