基于Arah6的花生间接竞争ELISA检测方法的建立  

A method for detecting peanut based on Ara h 6 by indirect competitive enzyme-linked immuno sorbent assay

作　　者：闫飞[1] 周宁菱[1] 罗春萍[2] 鲁俊华 李欣[1] 吴志华[1] 陈红兵[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,南昌大学中德联合研究院,江西南昌330047 [2]台州科技职业学院,浙江台州318020 [3]德胜中学,江西抚州344606

出　　处：《食品工业科技》2012年第17期303-306,共4页Science and Technology of Food Industry

基　　金：国家科技支撑计划(2011BAK10B03);国家自然科学基金(21162019);江西省科技支撑计划(2009BNA08100);南昌大学研究生创新专项基金项目

摘　　要：建立了基于花生主要过敏原蛋白Arah6的间接竞争酶联免疫检测法,实现了对食物中花生的定量检测。利用花生过敏原Arah6纯品免疫新西兰大白兔,制得兔抗Arah6的多克隆抗体,以Arah6纯品作为包被抗原、自制抗体为一抗、辣根过氧化物酶标记的羊抗兔IgG为二抗,确定了包被抗原质量浓度为1μg/mL,一抗最佳稀释度为1:50000,酶标二抗稀释度为1:5000。此检测方法对Arah6的定量检测范围为16.5～10000ng/mL(折合成含花生的含量,约为165ng/mL～100μg/mL),抑制方程为抑制率I=21.418lgC-6.0633(C为Arah6的质量浓度,单位ng/mL),IC50为414.6ng/mL,相关系数R2=0.9989。该检测方法灵敏度高,检测范围广,适用于食品中花生现场快速检测。An indirect competitive enzyme-linked immunosorbent assay（ ELISA） was developed to detect peanut in food,based on Ara h 6, a main allergen in peanut.The polyclonal antibody was prepared by immunizing New Zealand rabbits with Arah 6. Those parameters were optimized as following.The concentrations of the coating antigen,polyclonal antibody against Ara h 6 and horseradish peroxidase conjugated goat anti-rabbit IgG were 1μg/mL, 1：50000 and 1 ：5000 respectively.The inhibition equation of indirect competitive ELISA was obtained as I = 21.4181gC-6.0633 （I stand for inhibition ratio and C stand for mass concentration）and the curve was linear in range of 16.5-10000ng/mL Thus,this method has good sensitivity and wide range for the detection of Ara h 6 in peanut food,

关 键 词：花生过敏原 ARA h 6 多克隆抗体 间接竞争ELISA 检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]