Krambovitis E, Hatzidakis G, Hatzoglou A, Romain S, Durand A, Stefanakis A, Castanas E
Department of Applied Biochemistry and Immunology, Institute of Molecular Biology and Biotechnology, Crete, Greece.
Clin Chem. 1995 Jan;41(1):48-53.
A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturation of active receptors with their respective authentic ligands, (b) heat treatment of the cytosol to release the steroids from their cognate receptors before or after absorption with dextran-coated charcoal, and (c) measurement of both steroids present in the cytosol by a modified competitive-inhibition enzyme immunoassay. The useful range of the method was 10-4000 pmol/L for ER and 6.5-1000 pmol/L for PR. The correlation coefficient (r) between the one-point and Scatchard plot analysis was 0.95 for ER and 0.99 for PR. Comparison of the one-point ELIA and expected values with the radioligand binding assay (RLBA) results for EORTC samples gave r = 0.88 and 0.99 for ER and PR, respectively. Further comparison of the one-point ELIA with RLBA and with a commercial enzyme immunoassay, in blind testing of cancer tissue microsamples from 70 patients, gave good agreement for ER with r = 0.95-0.97 and concordance of 92.9-94.4% (cutoff, 15 pmol/g protein) against the other two methods. The results were more disperse in all three methods for PR estimation, the assay correlating perhaps better with the enzyme immunoassay (r = 0.90) at a concordance of 89.4% (same cutoff value).
本文描述了一种用于测定人乳腺癌组织微量样本中雌激素（ER）和孕激素（PR）受体的新方法（酶-配体免疫测定法，ELIA）。该技术基于对受体结合雌二醇和孕激素的非同位素测量，包括三个步骤：（a）用各自的天然配体同时饱和活性受体；（b）在使用葡聚糖包被活性炭吸附之前或之后，对胞浆进行热处理以从其同源受体中释放类固醇；（c）通过改良的竞争性抑制酶免疫测定法测量胞浆中存在的两种类固醇。该方法的有效范围对于ER为10 - 4000 pmol/L，对于PR为6.5 - 1000 pmol/L。单点分析与Scatchard图分析之间的相关系数（r）对于ER为0.95，对于PR为0.99。将单点ELIA和预期值与EORTC样本的放射性配体结合测定（RLBA）结果进行比较，ER和PR的r分别为0.88和0.99。在对70例患者的癌组织微量样本进行盲法检测时，将单点ELIA与RLBA以及一种商业酶免疫测定法进一步比较，结果显示ER的相关性良好，r = 0.95 - 0.97，与其他两种方法的一致性为92.9 - 94.4%（临界值，15 pmol/g蛋白质）。在所有三种PR测定方法中结果的离散度更大，该测定与酶免疫测定法的相关性可能更好（r = 0.90），一致性为89.4%（相同临界值）。
