直接竞争酶联免疫吸附法检测虾过敏蛋白

Direct competitive enzyme linked immunoassay for detection of shrimp allergic protein

目的:建立酶标抗原的直接竞争酶联免疫吸附法(dcELISA)检测食品中虾过敏蛋白,为食品过敏诊断试剂的开发和应用提供理论基础。方法:提取虾主要过敏蛋白,免疫小鼠制备抗虾过敏蛋白多克隆抗体,辣根过氧化物酶(HRP)标记抗原,建立酶标抗原的dcELISA检测虾过敏蛋白。结果:所建立的dcELISA法最低检测限为3.94ng/mL,标准曲线在0.12～128.86ng/mL范围内线性良好,批内和批间变异系数分别为6.16%和2.73%,回收率为82%～98%。结论:该方法具有良好的特异性、敏感性和稳定性,为进一步研制检测虾过敏蛋白的ELISA试剂盒提供有效的方法。

Objective：Direct competitive enzyme linked immunoassay（dcELISA） was established with enzyme-labeled antigen for detecting shrimp allergic protein in foods.Method：The main allergic protein of shrimp was extracted,and immunized mice to prepare polyclonal antibody,horseradish peroxidase（HRP） labeled antigen was also prepared.The dcELISA method was established for detecting shrimp allergic protein.Results：The results indicated that the LOD of the dcELISA was 3.94ng/mL,linear range of the standard curve was 0.12～128.86ng/mL,and the CV of intra-assay and inter-assay were 6.16% and 2.73% respectively.Recovery ranged from 82% to 98%.Conclusion：The method showed better specificity,sensitivity and stability,applying basis for further development of ELISA kit for shrimp allergic protein detection.