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目的分析家蚕过敏的儿童血清IgE与蚕蛹过敏原反应的图谱。方法提取蚕蛹的蛋白质成分,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对粗提液的蛋白质组分进行分析;用免疫印迹法(Western blotting)对蚕蛹过敏原成分进行鉴定;用离子交换层析对其过敏原进行初步纯化并进行免疫印迹鉴定。结果蚕蛹有20余种不同的蛋白质条带,其中最多的蛋白质主带集中在30、28、14、12ku。免疫印迹结果显示,1条18ku的条带为反应频率最高的过敏原(5/11)。经过离子交换层析,发现反应的主要过敏原在离子交换层析的第3个洗脱峰。结论儿童家蚕过敏患者血清IgE的反应图谱与成人有所不同,可为家蚕过敏诊断及治疗提供理论依据。
Objective To analyze the relationship between serum IgE and silkworm pupa allergen response in silkworm allergy. Methods The protein components of silkworm pupae were extracted and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein components of crude extracts were analyzed by Western blotting. Identification; ion exchange chromatography for its preliminary purification of allergens and Western blot identification. Results The silkworm pupae had more than 20 kinds of different protein bands, most of which were concentrated in 30,28,14,12ku. Immunoblotting showed that one 18ku band was the allergen with the highest frequency of response (5/11). After ion exchange chromatography, the major allergen of the reaction was found to be the third eluting peak in ion exchange chromatography. Conclusion The serum IgE response in children with silkworm allergy is different from that in adults and may provide a theoretical basis for the diagnosis and treatment of silkworm allergy.