荞麦过敏原特异性抗体酶联免疫吸附法检测试剂制备  

Preparation of an ELISA Method for Determining Buckwheat Allergen

作　　者：张昕[1] 崔晓东[2] 王转花[2] 

机构地区：[1]山西大学环境科学与工程研究中心,山西太原030006 [2]山西大学生物技术研究所,山西太原030006

出　　处：《食品科学》2011年第20期327-330,共4页Food Science

基　　金：山西省科技攻关项目(20100321101);太原市技术创新计划项目(2010-09)

摘　　要：以天然苦荞过敏蛋白包被聚苯乙烯微孔板,以鼠抗人IgE抗体标记辣根过氧化物酶,以间接酶联免疫吸附法(indirect enzyme-linked immunosorbent assay,IELISA)检测病人血清中的特异性抗体,分析并建立关键操作步骤及反应条件。结果表明:ELISA反应的最适工作条件为包被的抗原质量浓度100μg/mL、阳性血清稀释度1:50、酶标二抗稀释度1:1000,37℃时封闭的最佳时间为40min,包被抗原与阳性血清中抗体的最佳作用时间为40min,阳性血清与酶标二抗的最佳作用时间为40min。本方法具有良好的特异性、重复性和精密度。An indirect enzyme-linked immunosorbent assay（IELISA） was proposed to detect specific antibodies in tartary buckwheat allergenic patients,sera using polystyrene micro-well plate coated with natural tartary buckwheat allergenic proteins and horseradish peroxidase（HRP）-labeled mouse anti-human IgE.Key operations and reaction conditions were optimized.The results showed that the optimal coating antigen concentration,positive serum dilution,HRP-labeled mouse anti-human IgE concentration,blocking time at 37 ℃,reaction time between coating antigen and antibodies in positive sera,and reaction time between positive sera and HRP-labeled mouse anti-human IgE were 100 μg/mL,1：50,1：1000,40 min,40 min and 40 min,respectively.This method proved highly specific,repeatable and sensitive.

关 键 词：苦荞 过敏蛋白 试剂盒 间接酶联免疫吸附法 

分 类 号：R392.11[医药卫生—免疫学]