人血清转铁蛋白酶联免疫测定法的研究  

A STUDY ON QUANTITATION OF HUMAN SERUM TRANSFERRIN BY COMPETITIVE ENZYME IMMUNOASSAY

作　　者：何涛[1] 吴明泸[1] 罗文炽[1] 杨烨[1] 刘友平[1] 周志远[1] 

机构地区：[1]泸州医学院生物化学教研室,646000

出　　处：《泸州医学院学报》1999年第2期99-101,共3页Journal of Luzhou Medical College

摘　　要：目的：建立人血清转铁蛋白（Tf）的竞争性酶联免疫测定法（CEIA），促进Tf的实验研究及临床检测。方法：采用纯化人Tf免疫家兔制备抗血清，进行间接竞争性酶联免疫抑制试验，并进行灵敏度、精确度准确性、特异性试验。结果：通过棋盘滴定确定抗原包被浓度以10μg／ml，抗血清稀释度以1：4000为最佳工作条件，标准曲线工作范围为0．5～256μg／L，灵敏度为0．39μg／L，批内及批间变异系数分别为6．0％及8．2％，回收率为95．6％，与人血清白蛋白、牛血清白蛋白、铁蛋白、γ－球蛋白无交叉结合反应。用该法测定正常人（20例）、肝病患者（15例）血清Tf含量分别为2．61±0．37g／L、1．85±0．31g／L。结论：该法具有简便、准确、特异、灵敏和成本低的优点，可用于Tf的实验与临床研究。Objective: To develop a competitive enzyme immunoassay for the measurement of human serum transferrin (Tf) to facilitate the further strdy on Tf. Methods: The antibodies of Tf were prepared by immunizing rabbit. The indirect competitive inhibitory test was carried out. The optimum coating concentaion and antiserum titre were determinated by the checkerboard test. Its sensitivity, precision, recovery and specificity were also tested.Serum samples were taken from 20 normal humam subjects and 15 hepatitis patients for Tf measurement. Results: The optimum coatinng concentration and antiserum titer were 10 μg／ml and 1: 4000 respectively. The standard curve with a working range of 0. 5～256μg／L was plotted. The assay provided a sensitivity of 0. 36μg／L, the average intra-and inter-assay coefficient of variation were 6 .0% and 8. 2% respectively. The recovery of the assay was 95. 6%. No cross reactivittes with human serum albumin, bovin serum albumin, ferritin and γ- globulin were observed. The mean serum concentration of Tf in normal human subjects and hepatitis patients were 2. 61±0. 37g／L and 1. 85 ±0. 31g／L respectively. Conclusions: The competitive enzyme immunoassay we established is simple, precise, specific and of lower cost, which can be used in the experimental and clinical study on transferrin.

关 键 词：转铁蛋白 ELISA 血清 

分 类 号：R446.112[医药卫生—诊断学] R446.61[医药卫生—临床医学]