Ohtaki S, Endo Y
Nihon Naibunpi Gakkai Zasshi. 1979 Dec 20;55(12):1558-69. doi: 10.1507/endocrine1927.55.12_1558.
Enzyme-linked sandwich immunoassays for the measurement of thyroglobulin and anti-thyroglobulin autoantibody in human serum using silicone rod and beta-D-galactosidase were studied. These methods showed excellent results in specificity, sensitivity, precision and clinical application. 1) A method using silicone rod coated with rabbit (anti-human thyroglobulin) immunoglobulin G and rabbit (anti-human thyroglobulin) monovalent fragment of immunoglobulin G (Fab') conjugated with beta-D-galactosidase was developed for the measurement of circulating thyroglobulin. The sensitivity of the assay with as little as 2 microliter of serum was 10.7 amoles/tube corresponding to 3.5 ng/ml of serum, which was equal to or rather higher than that of radioimmunoassay. The correlation coefficient between values determined by the present assay and a double-antibody radioimmunoassay was 0.99 (n = 63, p less than 0.001). Circulating thyroglobulin was detectable in 90% of 146 normal subjects, the concentration being 13.3 +/- 10.3 ng/ml (mean +/- S.D.). Interference of anti-thyroglobulin autoantibody with the assay for thyroglobulin was smaller than that in radioimmunoassay. 2) Another method using human thyroglobulin conjugated with beta-D-galactosidase and silicone rod coated with human thyroglobulin was developed for the measurement of circulating (anti-human thyroglobulin) autoantibody. The sensitivity of the assay with as little as 5 microliter of serum was 7 fmoles/tube corresponding to 220 ng/ml of serum, which was equal to or rather higher than that of radioimmunoassay. The highly significant correlation was observed between the concentrations of anti-thyroglobulin autoantibody determined by the present assay and a radioimmunoassay (r = 0.80, n = 74, p less than 0.001) and also between those by the present assay and those by tanned red cell hemagglutination (r = 0.78, n = 199, p less than 0.001). No effect of thyroglobulin on the present assay was observed unless the ratio of the amount of thyroglobulin to that of (anti-human thyroglobulin) immunoglobulin G was higher than a tenth.
研究了使用硅棒和β-D-半乳糖苷酶测定人血清中甲状腺球蛋白和抗甲状腺球蛋白自身抗体的酶联夹心免疫分析法。这些方法在特异性、灵敏度、精密度和临床应用方面均显示出优异的结果。1）开发了一种方法，使用包被有兔（抗人甲状腺球蛋白）免疫球蛋白G和与β-D-半乳糖苷酶偶联的兔（抗人甲状腺球蛋白）免疫球蛋白G单价片段（Fab'）的硅棒来测定循环甲状腺球蛋白。该检测方法对低至2微升血清的灵敏度为10.7阿托摩尔/管，相当于血清3.5纳克/毫升，与放射免疫分析法相当或更高。本检测方法与双抗体放射免疫分析法测定值之间的相关系数为0.99（n = 63，p < 0.001）。在146名正常受试者中，90%可检测到循环甲状腺球蛋白，浓度为13.3±10.3纳克/毫升（平均值±标准差）。抗甲状腺球蛋白自身抗体对甲状腺球蛋白检测的干扰小于放射免疫分析法。2）开发了另一种方法，使用与β-D-半乳糖苷酶偶联的人甲状腺球蛋白和包被有人甲状腺球蛋白的硅棒来测定循环（抗人甲状腺球蛋白）自身抗体。该检测方法对低至5微升血清的灵敏度为7飞摩尔/管，相当于血清220纳克/毫升，与放射免疫分析法相当或更高。本检测方法与放射免疫分析法测定的抗甲状腺球蛋白自身抗体浓度之间观察到高度显著的相关性（r = 0.80，n = 74，p < 0.001），本检测方法与鞣酸红细胞血凝试验测定的浓度之间也观察到高度显著的相关性（r = 0.78，n = 199，p < 0.001）。除非甲状腺球蛋白与（抗人甲状腺球蛋白）免疫球蛋白G的量之比高于十分之一，否则未观察到甲状腺球蛋白对本检测方法的影响。
