天津医药

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牛奶过敏患者血清特异性抗体检测方法的建立※

常艳敏1,陈寅2,林书祥3,韩煦1,李会强4   

  1. 1. 天津南开医院检验科
    2. 武警云南总队医院
    3. 天津市儿童医院
    4. 天津医科大学免疫学教研室
  • 收稿日期:2012-01-13 修回日期:2012-04-16 出版日期:2012-09-15 发布日期:2012-09-15
  • 通讯作者: 李会强

The Establishment Determination Method for Milk Allergy Specific Antibody

  • Received:2012-01-13 Revised:2012-04-16 Published:2012-09-15 Online:2012-09-15

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摘要： 摘要 目的：建立双抗原夹心酶联免疫法测定疑似牛奶过敏患者血清特异性抗体。方法：用牛奶中主要过敏原包被酶标反应板同时标记辣根过氧化物酶制备酶标记抗原；待检血清中牛奶特异性抗体同时与包被抗原和酶标抗原结合，形成双抗原夹心复合物；酶催化底物显色终止反应后测定A450，以阴性对照A450均值2.1倍为临界点（Cut off）判定结果。结果：双抗原夹心法批间和批内精密度为3.7%-13.6%，敏感度为91.7%,特异度为94%,准确度为93.3%。结论：本文建立双抗原夹心酶联免疫法可以测定疑似牛奶过敏患者血清特异性抗体，用于牛奶过敏的体外诊断。

关键词: 牛奶过敏, 酶联免疫吸附试验, 双抗原夹心法, 特异性抗体

Abstract: Abstract Objective: to establish a double antigen sandwich enzyme linked immunosorbent assay (ELISA) to detect the specific antibody in milk allergic patient serum. Methods: main components of milk allergen were used to coat microwell plate and prepare enzyme conjugate antigen by labelling horseradish peroxidase (HRP). The specific antibody in test serum was combined with coating antigen and enzyme conjugate antigen simultaneously to form the double antigen sandwich compound. After enzyme catalyzed coloration and reaction termination, the absorbance at 450 nm (A450) was read and results were judged by setting 2.1 A450 of negative control as the cut off value. Results: the coefficient of variations (CV) in the group and between groups of the developed method were 3.7%-13.6%, and its sensitivity, specificity, accuracy were 91.7%, 94%, 93.3% respectively. Conclusion：the double antigen sandwich ELISA established in this study can detect the specific antibody in doubtful milk allergic patients serum，showing its feasibility in vitro diagnosis of milk allergy.

Key words: milk allergy, ELISA, double antigen sandwich method, specific antibody