Chemical and Pharmaceutical Bulletin
Online ISSN : 1347-5223
Print ISSN : 0009-2363
ISSN-L : 0009-2363
Fluorescence Enzyme Immunoassay of 17α-Hydroxyprogesterone in Dried Blood Samples on Filter Paper and Its Application to Mass Screening for Congenital Adrenal Hyperplasia
HIDETOSHI ARAKAWAMASAKO MAEDAAKIO TSUJIHIROSHI NARUSEEMIKO SUZUKIAKIRA KAMBEGAWA
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Keywords: peroxidase
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1983 Volume 31 Issue 8 Pages 2724-2731

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Abstract

An enzyme immunoassay of 17α-hydroxyprogesterone (17-OHP) in dried blood spotted onto filter paper has been developed. 17-OHP was conjugated to horseradish peroxidase by the mixed anhydride method. Separation of free and bound 17-OHP-peroxidase conjugate was done by the use of insolubilized antibody, prepared by coating polyacetal beads with purified immunoglobulin G (IgG) of goat anti-rabbit IgG serum. The enzyme activity was measured by the fluorophoto-metric method using 3-(p-hydroxyphenyl)propionic acid and H2O2 as substrates. The sensitivity of the present method was 1.0 pg/tube for 17-OHP. The intra-and interassay coefficients of variation were 5.4-7.0% and 8.3-14.7%, respectively. The present enzyme immunoassay method could be applied to mass screening for congenital adrenal hyperplasia.

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© The Pharmaceutical Society of Japan
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