基于化学发光酶免疫分析法检测花生过敏原Ara h 2  

Detection of peanut allergen Ara h 2 based on chemiluminescence enzyme immunoassay

作　　者：王耀[1] 郭开通 王成宾 宋乾召 李才云 葛梦鋆 蔡文锦 孙亚宁[2] 邢云瑞 胡骁飞[2] WANG Yao;GUO Kaitong;WANG Chengbin;SONG Qianzhao;LI Caiyun;GE Mengjun;CAI Wenjin;SUN Yaning;XING Yunrui;HU Xiaofei(Food and Bioengineering College,Henan University of Science and Technology/Henan International Joint Laboratory of Food Green Processing and Quality Safety Control,Luoyang 471003,China;Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

机构地区：[1]河南科技大学食品与生物工程学院,河南省食品绿色加工与质量安全控制国际联合实验室,河南洛阳471003 [2]河南省农业科学院动物免疫学重点实验室,河南郑州450002

出　　处：《食品与发酵工业》2024年第20期266-272,共7页Food and Fermentation Industries

基　　金：国家自然科学基金项目(31702218);河南省科技攻关项目(232102320298);河南省高校青年骨干教师培养计划(2023GGJS046);河南省研究生教育改革与质量提升工程项目(HNYJS2020 JD06);河南省自然科学基金(232300421274);河南省农业科学院自主创新项目(2023ZC086)。

摘　　要：Ara h 2是花生主要过敏原之一,为开发食物中Ara h 2过敏原成分的快速检测方法,减少因误食导致花生过敏事件的发生,该研究采用鼠源单克隆抗体作为捕获抗体、兔源多克隆抗体作为检测抗体,通过棋盘法优化抗体工作浓度,建立了一种检测花生过敏原Ara h 2的间接双抗夹心化学发光酶免疫分析法,并对该方法的灵敏度、准确度、精密度和特异性进行评价。该方法的检出限为1.085 ng/mL,线性范围为3.12~200 ng/mL,添加回收率为78.30%~94.39%,批内和批间变异系数均小于10%,且特异性良好,与其他常见食物过敏原无交叉反应。该方法与相同抗体所建立的间接双抗夹心酶联免疫吸附测定(enzyme-linked immunosorbent assay, ELISA)方法相比,在灵敏度上表现出一定优势。该研究开发的化学发光酶免疫分析法可对花生食品生产过程中和消费前的Ara h 2过敏原成分检测提供可靠的技术支持。Ara h 2 is one of the major peanut allergens.This study aimed to develop a rapid detection method for Ara h 2 allergens in food and reduce the occurrence of peanut allergy caused by accidental eating.In this study,a mouse monoclonal antibody was used as a capture antibody and a rabbit polyclonal antibody as a detecting antibody,and the working concentration of antibodies was optimized by the checkerboard method.An indirect double-antibody sandwich chemiluminescence enzyme immunoassay for the detection of peanut allergen Ara h 2 was established,and the sensitivity,accuracy,precision,and specificity of the method were evaluated.The detection limit of the method was 1.085 ng/mL,the linear range was 3.12-200 ng/mL,the recovery rate was 78.30%-94.39%,both intra-batch and inter-batch coefficients of variation remained below 10%,and the method demonstrated excellent specificity by exhibiting no cross-reactivity with other prevalent food allergens.In a comparative analysis with an indirect double-antibody sandwich ELISA method employing the same antibodies,this method displayed distinct advantages in terms of sensitivity.The chemiluminescent enzyme-linked immunosorbent assay developed in this study offers dependable technical support for the evaluation of Ara h 2 allergen components during peanut food production processes and prior to consumption.

关 键 词：化学发光酶免疫分析 花生过敏原 Ara h 2 检测 

分 类 号：TS2[轻工技术与工程—食品科学与工程]