AN ENZYME-IMMUNOASSAY FOR DETERMINING TOTAL THYROXINE IN HUMAN SERUM USING AN ULTRAMICROANALYTICAL SYSTEM
AN ENZYME-IMMUNOASSAY FOR DETERMINING TOTAL THYROXINE IN HUMAN SERUM USING AN ULTRAMICROANALYTICAL SYSTEM
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DOI：
10.1016/0009-8981(91)90137-2
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发表时间：
1991-03-29
影响因子：
5
通讯作者：
BLANCA, S
中科院分区：
文献类型：
--
作者：
GONZALEZ, RR;ROBAINA, R;BLANCA, S
一种简单、快速的竞争性超微量酶联免疫吸附试验已被开发用于测量总甲状腺素（T4），仅使用10 μ l的血清。 我们的新UME是基于4-甲基伞形酮基-β-D-吡喃半乳糖苷水解产物的荧光测量。 将T4-β-半乳糖苷酶缀合物和单克隆抗体固定在聚乙烯板上，水杨酸钠用作甲状腺素结合蛋白的封闭剂。 分析步骤使用名为“SUMA”（超微量分析系统）的半自动化分批测定系统进行。 T4测定在2 h内完成，测量范围为24-386 nmol/L。 试验内变异系数（CV）为4.6-6.9%，试验间C. V.为9.0-12.4%，取决于T4浓度。 回收率范围为99.2- 111%。 回归分析显示，与放射免疫法测定的结果有良好的相关性（n = 121，r = 0.946，P < 0.01）。
A simple, rapid competitive ultramicroElisa assay has been developed for the measurement of total thyroxine (T4) using only 10-mu-l of serum. Our novel UME is based on fluorescence measurements of the hydrolytic product of 4-methyl-umbelliferyl-beta-D-galactopyranoside. T4-beta-Galactosidase conjugates and monoclonal antibodies were immobilized on polyvinyl plates, with sodium salicylate used as a blocking agent for thyroxine binding protein. The analytical steps were carried out using a semiautomated batch-assay system entitled "SUMA" (system for ultramicroanalysis). The T4 assay was completed in 2 h, with a measuring range of 24-386 nmol/L. The intra-assay coefficient of variation (CV) was 4.6-6.9% and the inter-assay C.V. 9.0-12.4% depending on the T4 concentrations. Percentage recovery ranged from 99.2-111%. Regression analysis showed a good correlation with an established radioimmunoassay (n = 121, r = 0.946, P < 0.01).