应用SPA的ELISA检测血清胰岛素抗体

APPLICATION OF ELISA USING SPA TO DETECT INSULIN ANTIBODY OF SERUM

应用SPA代替抗人IgG第二抗体的ELISA测定血清胰岛素抗体(IA)。经正交试验分析,受检血清稀释度是影响IA测定的主要因素。选用胰岛素包被浓度为10μg/ml,受检血清1:2.5稀释时,IA测定可获较满意的结果。竞争性结合抑制试验表明IA测定具有特异性。血清稀释试验提示OD值大小反映了IA滴度高低。28例正常人血清IA测定结果OD正常值范围(X±2SD)为0.05～0.25。使用胰岛素的DM病例均为阳性,而未使用胰岛素病例未发现IA阳性。本文认为应用该法测定IA是一种稳定、特异、简便的方法。

This paper reports the study of application of ELISA using SPA instead of anti-human IgG as the second antibody to detect insulin antibody ( IA ) of serum. The assay conditions were evaluated by the Orthogonal Test which showed that serum dlution was the main factor which affected IA measurement. The results were pretty good when assay was carried out when insulin-coated concentration was 10 μg per ml, and serum dilution was 1 to 2.5. The binding-competitive inhibition test inlicates that the assay is specific for detection of IA. The serum dilution lest suggests that OD value .s able to reflect IA level of serum. On basis of our results, the normal range of OD value for IA is from 0.05 to 0.25(X±2SD). In our experiment all of the patients with diabetes mellitus who had been treated with insulin were IA-positive. However the patients who had not been treated with insulin were IA-negative. So it is considered that the assay reported here is useful for IA detection.