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Direct Solid-Phase Time-Resolved Fluoroimmunoassay of 17α-Hydroxyprogesterone in Dried Blood Spotted on Filter Paper

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Abstract

A direct, solid-phase time-resolved fluoroimmunoassay (TRFIA) for measuring 17α-hydroxyprogesterone (17-OHP) in dried blood spotted on filter paper is described. The assay is based on competition between europium (Eu)-labeled 17-OHP and free 17-OHP in blood specimens for polyclonal rabbit anti-17-OHP antibodies. The antibody-label complex is separated by binding to anti-rabbit antibodies coated onto microtiter plate wells. The amount of bound Eu-labeled 17-OHP is inversely proportional to the amount of 17-OHP in the sample. After separation of bound and free fractions, the fluorescence intensity of Eu is measured by TRFIA. 17-OHP values obtained by this method correlated well with those obtained by enzyme linked immunosorbent assay (ELISA) (r=0.969). The detection limit of 17-OHP was 1 pg/assay, being almost equivalent to those of the chemiluminescent and fluorescent enzyme immunoassays developed previously.

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Authors and Affiliations

  1. School of Pharmaceutical Science, Showa University, Hatanodai, Shinagawa, 142, Japan

    Masako Maeda, Kwang Hun Yang & Akio Tsuji

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  1. Masako Maeda
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  2. Kwang Hun Yang
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  3. Akio Tsuji
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Maeda, M., Yang, K.H. & Tsuji, A. Direct Solid-Phase Time-Resolved Fluoroimmunoassay of 17α-Hydroxyprogesterone in Dried Blood Spotted on Filter Paper. ANAL. SCI. 7, 699–702 (1991). https://doi.org/10.2116/analsci.7.699

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  • DOI: https://doi.org/10.2116/analsci.7.699

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