甲状腺素(T_4)酶联免疫测定法的研究——Ⅱ、T_4-EIA方法的建立

Ⅱ.Methodological investigation

本文介绍以辣根过氧化物酶(HRP)标记的 T_4为示踪物的液相酶酸免疫测定方法的建立及方法学考核。加 ANS 阻断血样品(20μl)中之 T_4与 TBG 的结合,然后直接进行 EIA。以驴抗兔血清为分离剂,邻苯二胺为底物溶液。方法之特异性,准确性,一致度及灵敏度(0.62μg/dl)均能满足临床需要。还介绍了 ANS 浓度、B-F 分离手段及降低非特异性结合的一些实践经验。EIA 方法简便、快速,不需特殊设备又无放射性污染,肯定会迅速发展及推广应用。

The present method was developed and used for the determination of serum T_4concentration with the horse-radish peroxidase labelled T_4 as the tracer,T_4 antiserum,horse-radish peroxidase labelled T_4 and T_4 standard solutionwere all prepared in this laboratory.Methodological examination was performed.including sensitivity or minimum detectability (1.25%μg/dl serum),percisionor intrabatch and interbatch variations (5.3 and 8.2%,respectively),averagerecovery (99.6%) and parallel test.i.e.construction of the T_4-content curveswere prepared from hyperthyroid serum of various dilutions