Kohno H, Sueshige N, Oguri K, Izumidate H, Masunari T, Kawamura M, Itabe H, Takano T, Hasegawa A, Nagai R
Fuji Research Laboratories, Kyowa Medex Co., Ltd., Nagaizumi-cho, Shizuoka, Japan.
Clin Biochem. 2000 Jun;33(4):243-53. doi: 10.1016/s0009-9120(00)00065-5.
To develop a simple and practical enzyme immunoassay (EIA) for oxidatively modified low density lipoprotein (Ox-LDL) in human blood, a biological marker of atherogenesis.
A sandwich EIA suitable for the measurement of human Ox-LDL was developed using the mouse monoclonal antibody FOH1a/DLH3. This antibody, specific for oxidized phosphatidylcholine, was used as the capture antibody, and a horseradish peroxidase (HRP)-labeled goat anti-human apolipoprotein-B (Apo-B) IgG was used for detection. Copper-oxidized human LDL, prepared under controlled conditions, was used as a standard and the results of the EIA were expressed in arbitrary units (U/mL).
This EIA meets all the requirements for use in routine clinical assays in terms of sensitivity (detection limit: 1 U/mL), reproducibility (total CV: 2.3-7.7%), accuracy (recovery: 90.6-103.8%), simplicity and rapidity (<4 h). Clinical performance of the assay was assessed by measurement of the Ox-LDL in the plasma of normal subjects (10.8 +/- 2.8 U/mL, mean +/- SD) and patients with coronary heart disease (CHD) (19.7 +/- 10.2). The present EIA had a sensitivity of 79% and a specificity of 75% for CHD.
We have developed a new assay, suitable for the measurement of Ox-LDL in human blood, which meets the requirements for routine clinical assay.
开发一种简单实用的酶免疫分析法（EIA），用于检测人血液中氧化型低密度脂蛋白（Ox-LDL），这是动脉粥样硬化发生的一种生物标志物。
使用小鼠单克隆抗体FOH1a/DLH3开发了一种适用于检测人Ox-LDL的夹心EIA。这种对氧化磷脂酰胆碱具有特异性的抗体用作捕获抗体，辣根过氧化物酶（HRP）标记的山羊抗人载脂蛋白B（Apo-B）IgG用于检测。在受控条件下制备的铜氧化人低密度脂蛋白用作标准品，EIA结果以任意单位（U/mL）表示。
该EIA在灵敏度（检测限：1 U/mL）、重复性（总变异系数：2.3 - 7.7%）、准确性（回收率：90.6 - 103.8%）、简便性和快速性（<4小时）方面满足常规临床检测的所有要求。通过测量正常受试者（10.8±2.8 U/mL，均值±标准差）和冠心病（CHD）患者（19.7±10.2）血浆中的Ox-LDL来评估该检测方法的临床性能。本EIA对冠心病的灵敏度为79%，特异性为75%。
我们开发了一种新的检测方法，适用于检测人血液中的Ox-LDL，满足常规临床检测的要求。
