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目的:鉴定河虾中的过敏原组分,对主要过敏原组分进行纯化并分析其致敏性强弱。方法:用磷酸盐缓冲液(PBS)制备河虾蛋白粗提液,将其与11例虾过敏症患者血清IgE进行Westernblot,鉴定各种分子量的河虾过敏原组分;河虾蛋白粗提液用硫酸铵沉淀、G-50凝胶层析和阴离子交换层析等方法纯化其主要过敏原组分,再用虾过敏患者血清IgE进行Westernblot鉴定;并将纯化的相对分子质量(Mr)为21000、36000、800003种主要过敏原组分与虾过敏症患者血清IgE做间接ELISA,以分析其致敏性强弱。结果:West-ernblot结果显示,河虾蛋白粗提液出现9个阳性条带;其中3种主要过敏原组分21000、36000、80000与患者血清的反应率为36.4%,63.6%,45.5%;将这3种Mr的蛋白组分分别做间接ELISA,结果显示21000、36000、80000过敏原组分与11例虾过敏症患者的混合血清IgE结合的吸光值都显著高于河虾蛋白粗提液。结论:河虾中至少存在9个过敏原组分;21000、36000、8000等3种蛋白组分为河虾的主要过敏原组分,其中以36000过敏原组分致敏率和致敏性最强。进一步研究将探明21000、36000、80000等3种河虾过敏原组分的共同抗原表位,以期为明确食物过敏原检测、临床诊断和虾过敏原疫苗设计提供基础。
OBJECTIVE: To identify allergen components in shrimp, to purify the major allergen components and to analyze their allergenicity. Methods: The crude shrimp protein extract was prepared by phosphate buffered saline (PBS), which was then subjected to Western blot with serum IgE from 11 cases of shrimp allergy patients to identify shrimp allergen components of various molecular weights. The major allergen components were purified by ammonium sulphate precipitation, G-50 gel chromatography and anion exchange chromatography, and then identified by Western blot with serum IgE from shrimp allergy sufferers. The relative molecular mass (Mr) was 21000 , 36000,800003 kinds of major allergen components and shrimp allergy patients serum IgE indirect ELISA to analyze its sensitivity. Results: West-ernblot results showed that there were 9 positive bands in crude protein extracts of Rhizopus sp., Of which the reaction rates of 21000, 36000 and 800000 for three major allergens were 36.4%, 63.6% and 45.5% respectively. The three kinds of Mr protein components were indirect ELISA results showed that 21000, 36000, 80 000 allergen components and 11 cases of shrimp allergy patients mixed serum IgE binding absorbance values were significantly higher than shrimp protein crude extract . Conclusion: There are at least 9 allergen components in shrimp; 21000, 36000, 8000 and other protein components are the main allergen components of shrimp, of which 36000 allergen component sensitization rate and the most sensitive Strong. Further research will identify common antigenic epitopes of 21000, 36000, 800000 and other three kinds of shrimp allergen components in order to provide a basis for the clear detection of food allergens, clinical diagnosis and shrimp allergen vaccine design.