直接竞争酶联免疫吸附法检测虾过敏蛋白

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。

Direct competitive enzyme linked immunoassay for detection of shrimp allergic protein

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.