Development of flow injection system for detection of allergen using fluorochrome.

许多免疫测定方法已经被开发出来用于传感或检测过敏原。皮肤试验包括点刺试验和划痕试验。然而，这些测试是危险的，因为过敏反应的血清抗体。因此，临床和食品工业领域都需要一种安全、简单、快速的检测过敏反应的方法。我们开发了一种自动流动免疫分析系统，使用离子交换柱检测食物过敏原。根据等电点的差异，利用离子交换柱将抗原-抗体复合物从未反应的分析底物中分离出来。本研究构建了由免疫反应柱、离子交换柱和检测器组成的过敏原检测系统。荧光强度与变应原浓度在0.01 ~ 2.0 mg/ml范围内呈线性关系。在此范围内相关系数为0.994。该方法检测速度快（10 min），操作简单，比使用微量滴度板的传统ELISA检测精度高（<2%）。游离荧光染料共轭IgE的重复利用是可能的。我们还构建了一种新的自动流式免疫测定系统，用于定量系统性红斑狼疮患者血清中的抗双链DNA自身免疫抗体。在0 ~ 300马克杯/毫升范围内，信号与抗dsdna抗体浓度呈线性关系。该技术允许在25分钟内检测抗dsdna自身免疫抗体。这些简单、快速的免疫分析方法可用于过敏原的连续检测。

Many immunoassay methods have been developed for sensing or detecting allergens. The skin tests including prick test and scratch test are typical methods. However, the these tests are dangerous because of anaphylaxis by serum antibodies. Therefore, a safe, simple and rapid method for detecting allergic reaction is required in the clinical and food industry fields. We have developed an automated flow immunoassay system using an ion-exchange column for the detection of food allergens. Antigen-antibody complexes were subsequently separated from unreacted analytical substrate by ion-exchange column, on the basis of the difference in isoelectric point, In this study, the allergen detection system equipped with an immunoreaction column, an ion-exchange column and a detector was constructed. A linear relationship was obtained between the fluorescence intensity and allergen concentration in the range of 0.01 2.0 mg/ml. The correlation coefficient was 0.994 within this range. This method was faster (10 min) and simple to use, and gives higher precision (<2%) than the conventional ELISA using microtiter plates. Reuse of free fluorochrome conjugated IgE was possible. We also constructed a novel automated flow immunoassay system for quantification of anti-double-stranded (anti-ds) DNA autoimmune antibodies in the serum of patients suffering from systemic lupus erythematosus. The assay yielded a linear relationship between signal and concentration of anti-dsDNA antibody in the range of 0 300 mug/ml. This technique permits the assay of anti-dsDNA autoimmune antibodies within 25 min. These simple and rapid immunoassay methods can be used for the continuous detection of allergen.