ABC-ELISA法检测牛乳过敏小鼠模型中的特异性IgE  

Experimental Study on Detection of the Specific IgE in Sera of the Milk Allergen Model Mice by ABC-ELISA

作　　者：毛露甜[1] 向军俭[2] 张在军[2] 

机构地区：[1]惠州学院生命科学系,广东惠州516007 [2]暨南大学生命科学技术学院分子免疫学与抗体工程研究中心,广东广州510632

出　　处：《生物技术通讯》2006年第3期389-391,共3页Letters in Biotechnology

基　　金：广东省自然科学基金项目[2003(36708)];教育部重点项目[2003(03166)]

摘　　要：目的:建立并优化一种有效检测特异性IgE的方法,以取代皮内试验快速诊断过敏症。方法:用乳清蛋白皮下注射BALB/c小鼠建立牛奶过敏模型,获得小鼠抗血清进行方法学研究。结果:乳清蛋白的最适包被量为500ng,用pH9.4的碳酸盐缓冲液作为包被液,4℃包被过夜效果最佳,选用0.2%的白明胶/1%的BSA封闭效果最好,生物素化抗鼠IgE抗体的最佳反应浓度为1∶1000。结论:建立了检测特异性IgE的生物素-亲和素系统改良的ABC-ELISA法,该法特异、灵敏、安全、简便,可用于临床诊断过敏症。Objective： To develop and optimize the assay of detect specific IgE effectively which would be used to substitute for the prick test in clinic allergies diagnosis. Methods： Allergic BALB/c model of whey protein had been established by hypodermic injected with the adjuvant of aluminum hydroxide, then anti-serum with high titer of slgE was isolated for detection. Results： In ABC-ELISA, the optimal contents of whey protein for coating was 500 ng, which was incubated overnight at 4℃ in carbonate-buffered saline（pH9.4）, then blocked with 0.2% gelatin/1% BSA at 37℃ for 2 hours. After washes with PBST, the blots were incubated with biotin-rat anti mouse IgE heavy chain as secondary antibody, which the optimal reaction content was 1：1 000. Conclusion： A reformed system of biotin and conjugated streptomyces avidin had been established, which was specific, sensitive, safe and easy. It would be a new technique in clinic allergies diagnosis.

关 键 词：ABC-ELISA 食物过敏 特异性IGE 

分 类 号：R392.11[医药卫生—免疫学]