间接竞争ELISA法检测坚果类过敏原特异性研究  

The specific detection research of nuts allergen with indirect-competitive ELISA

作　　者：范建涛 张爱琳[1,2] 姚尧[1] 张钰鑫[1] 苗颖[1,2] 

机构地区：[1]天津农学院食品科学与生物工程学院,天津300384 [2]天津市农副产品深加工技术工程中心,天津300384

出　　处：《食品工业科技》2015年第24期64-66,70,共4页Science and Technology of Food Industry

基　　金：中国博士后基金面上项目(2013M541181);2014年大学生国家创新项目(201410061071);天津农学院大学生科技创新项目

摘　　要：以腰果蛋白为过敏抗原,腰果蛋白致敏的阳性小鼠血清作为一抗,羊抗鼠Ig E-HRP为二抗建立双抗体间接酶联免疫吸附实验(ELISA)体系,通过棋盘法确定腰果蛋白最佳反应条件工作浓度为15.92μg/m L,最佳一抗稀释度均为1∶400,最佳酶标二抗的稀释度为1∶1000,腰果蛋白浓度为1.99μg/m L。并用该方法分别对腰果、核桃、榛子、开心果、板栗、杏仁等六种坚果进行检测分析,未发现有交叉反应,本实验建立的间接竞争性ELISA诊断方法具有良好的敏感性和特异性,为坚果过敏性食品的监测及时而准确的诊断奠定了基础。The methods of the indirect competitive ELISA for determining allergen in nuts were developed by using the self-prepared positive mice serum as a resistance in Cashew protein sensitization. The immunoglubing Ig E- HRP for 2 resistance was adopted. Cashew protein was identified through checkerboard microlilution method,the optimal solution was 15.92 μg/m L,the equal dilution multiples of mice Ig E and antibody-HRP were400 and 1000,cashew protein concentration was 1.99 μg/m L. Using the method of cashew nuts and walnuts,hazelnuts,pistachios,chestnut,almond nuts,six test cross reaction in nuts analysis had not been found. The method for rapid detection to allergen about nuts was established firstly by using and sensitivity and reliability,accurately specificity.

关 键 词：坚果 过敏原 ELISA 棋盘法 

分 类 号：TS201.1[轻工技术与工程—食品科学]