甲状腺刺激ホルモンの酵素免疫測定法の開発と臨床応用

概要

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An enzyme-immunoassay for the thyroid-stimulating hormone (TSH) was developed through the use of horseradish peroxidase as a label enzyme. TSH-peroxidase conjugate was prepared according to the periodate oxidation method. Procedures were established using the conjugate and an insoluble second antibody coupled with activated Sepharose 4B. Enzyme activity was determined by the fluorescence reaction of tyramine and hydrogen peroxide as substrates. The sensitivity of the method was sufficiently satisfactory to determine serum TSH concentrations as 0.06μU/tube, which is similar to the results of a radioimmunoassay. TSH values obtained in an unknown 84 samples by the method were in good agreement with those obtained by radioimmunoassay. The coefficients of variation were 1.8 to 5.3% (within assay) and 5.1 to 22% (between assays).<BR>Interference of serum, hemolysis and eluate from dried blood spots using the assay system were also examined, and the applicability of the method in clinical practice was discussed. Since the present method requires only minimal equipment and carries no radiation hazards, it would be useful for routine assays of TSH. Though the method requires further study, it is expected that the method will become an important tool especially in the field of neonatal mass-screening for congenital hypothyroidism.
日本内分泌学会の論文