天津医药

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虾有效致敏组分的提取及在sIgE检测中的初步应用*

任杰1,单立新2,陈凯3,李韶深4,李会强5   

  1. 1. 天津医科大学
    2. 天津市北辰医院
    3. 天津市北辰医院检验科
    4. 天津市中医药研究院附属医院
    5. 天津医科大学免疫学教研室
  • 收稿日期:2013-11-25 修回日期:2014-01-17 出版日期:2014-05-15 发布日期:2014-05-15
  • 通讯作者: 任杰

Extraction the Effective Component of Shrimp and the Application of Detection sIgE

2,kai chen 2,   

  • Received:2013-11-25 Revised:2014-01-17 Published:2014-05-15 Online:2014-05-15

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摘要： 目的 通过改善虾类包被抗原质量，提高血清sIgE检测的敏感性。方法 制备中华对虾蛋白溶液，经葡聚糖凝胶层析初步分离大于55KD蛋白组分，经SDS-PAGE技术分析其蛋白组成，采用酶免疫印迹和酶联免疫法初步评价纯化后抗原是否能够提高血清sIgE检测结果的敏感性。结果：葡聚糖凝胶层析法可初步分离>55kD的蛋白组分，其中主要包含10条明显蛋白条带；如以纯化后蛋白作为包被抗原，斑点酶免疫印迹显示，可提高弱阳性血清斑点光密度。同时，两种包被抗原的酶联免疫吸附试验结果显示，虾有效致敏成分作为包被抗原时，92.9%的虾过敏患者血清sIgE检测值升高。结论 采用中华对虾有效致敏原组分作为虾过敏检测抗原，有望提高血清sIgE检测结果的敏感性。

关键词: 食物过敏, 虾, 特异性IgE, 免疫印迹, 敏感性。

Abstract: Objective To improve the detection sensitivity of serum sIgE by improve the quality of shrimp coated antigen. Methods The shrimp protein extracts were prepared and those >55KD proteins were separated by Gel Filtration Chromatography. The separated protein components were separated by SDS–PAGE and using Western blot and ELISA to evaluate whether the detection sensitivity of serum sIgE could be improved by Purified antigen.Result Those >55KD proteins ccould be successfully separated by Gel Filtration Chromatography. The SDS-PAGE of those >55KD proteins showed 10 identifiable bands. Dot-pot immunoassay showed that >55KD proteins could be improved the density spots of the weak serum. Meanwhile, the result of ELISA showed that 92.9% detection value of serum sIgE increased by coating effctive antigen. Conclusions The sensitivity of detection for serum sIgE might be improved by effective protein components of shrimp as coated antigen.

Key words: food allergy, shrimp, sIgE, Western blot, sensitivity.