More sensitive and simpler immune complex transfer enzyme immunoassay for antithyroglobulin igg in serum
More sensitive and simpler immune complex transfer enzyme immunoassay for antithyroglobulin igg in serum
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更灵敏、更简单的免疫复合物转移酶免疫分析法测定血清中的抗甲状腺球蛋白 Igg
DOI：
10.1002/jcla.1860030306
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发表时间：
1989
影响因子：
2.7
通讯作者：
E. Ishikawa
中科院分区：
文献类型：
--
作者：
T. Kohno;T. Mitsukawa;S. Matsukura;Y. Tsunetoshi;E. Ishikawa
本文介绍了一种更灵敏、更简便的血清抗甲状腺球蛋白抗体免疫分析方法，并将其应用于健康人和甲状腺疾病患者的抗甲状腺球蛋白抗体检测。用二硝基苯基甲状腺球蛋白和甲状腺球蛋白-β-D-半乳糖苷酶结合物与受试者血清中的抗甲状腺球蛋白Ig G反应。将抗甲状腺球蛋白Ig G、二硝基苯基甲状腺球蛋白和甲状腺球蛋白-β-D-半乳糖苷酶结合物包被亲和纯化的兔抗甲状腺球蛋白免疫球蛋白包被在两个聚苯乙烯小球上。用洗涤的方法去除试验血清中的非特异性免疫球蛋白，用二硝基苯基-L-赖氨酸洗脱，转移到包被亲和纯化的兔抗人免疫球蛋白γ链的两个聚苯乙烯小球中。以4-甲基伞形花环-β-D-半乳糖苷为底物，用荧光法测定了β-D-半乳糖苷酶与聚苯乙烯小球的结合活性。与以往的免疫复合物转移酶免疫分析方法相比，简化了一步孵育步骤和一步洗涤步骤，血清中抗甲状腺球蛋白Ig G的检出限(0.1μg/L)降低了1 0倍。与以前的研究相比，更仔细和更广泛的检查显示，在很大一部分健康受试者以及所有Graves病和慢性甲状腺炎患者中都存在抗甲状腺球蛋白抗体。
A more sensitive and simpler immune complex transfer enzyme immunoassay for anti‐thyroglobulin IgG in serum and its use for the measurement of anti‐thyroglobulin IgG in healthy subjects and patients with thyroid diseases are described. Antithyroglobulin IgG in test serum was reacted simultaneously with dinitrophenyl thyroglobulin and thyroglobulin‐β‐D‐galactosidase conjugate. The complex formed of antithyroglobulin IgG, dinitrophenyl thyroglobulin, and thyroglobulin‐β‐D‐galactosidase conjugate was trapped onto two polystyrene balls coated with affinity‐purified rabbit (antidinitrophenyl bovine serum albumin) IgG. The polystyrene balls were washed to eliminate nonspecific IgG in the test serum, and the complex was eluted from the polystyrene balls with dinitrophenyl‐L‐lysine and transferred to two polystyrene balls coated with affinity‐purified rabbit (anti‐human IgG γ‐chain) IgG. β‐D‐Galactosidase activity bound to the polystyrene balls was assayed by fluorimetry using 4‐methylumbelliferyl‐β‐D‐galactoside as substrate. As compared with the previous immune complex transfer enzyme immunoassay, the procedure was simplified by reducing one incubation step and one washing step, and the detection limit of antithyroglobulin IgG in serum (0.1 μg/liter) was lowered 100‐fold. More careful and extensive examination than in a previous study revealed the presence of antithyroglobulin IgG in a large proportion of healthy subjects and in all patients with Graves' disease and chronic thyroiditis.