基于虾类原肌球蛋白共同表位肽抗体的过敏原检测方法
目的 建立一种能够同时检测多种虾原肌球蛋白的方法。方法 通过合成不同虾类的共同表位肽, 制备能够识别多种虾类原肌球蛋白的共同表位肽多克隆抗体, 建立快速灵敏的虾类原肌球蛋白酶联免疫检测方法。结果 该检测方法在4.79~1400 ng/mL的浓度范围内线性关系良好, 其线性回归方程为Y=-19.083X+ 98.303(R2=0.9813), 最低检出限(IC10)为1.85 ng/mL; 样品加标回收率在94.37%~103.45%之间; 该检测方法与软体动物的原肌球蛋白有交叉反应, 与鱼肉蛋白、牛奶、花生蛋白等无交叉反应; 批内变异系数为3.4%~9.1%, 板间变异系数为12.9%~19.6%, 贮藏试验显示该酶联免疫试剂盒可以在4 ℃下保存6个月以上, 能够应用于食品中虾类过敏原的检测。结论 采用共同表位抗体, 成功建立了基于酶联免疫的过敏原检测方法。
Objective To establish a method for simultaneous determination of various shrimp tropomyosins. Methods By synthesis of common peptide epitopes from different kinds of shrimp, polyclonal antibodies were produced which could identify a variety of shrimp tropomyosins. A rapid and sensitive ELISA method was established for the detection of shrimp tropomyosins. Results The method had a good linear relationship in the range of 4.79~1400 ng/mL, and the linear regression equation was Y=-19.083X+98.303(R2=0.9813). The limit of detection was 1.85 ng/mL(IC10), while the recovery rates were 94.37%~103.45%. The detection method was cross-responsive with invertebrate tropomyosin and had no cross reaction with fish protein, milk, and peanut protein. The coefficients of variation of the batch were 3.4%~9.1%, and the coefficients of variation of the plates were 12.9%~19.6%. The storage test showed that the ELISA kit could be stored more than 6 months at 4 ℃, which could be applied for the detection of shrimp allergens in food. Conclusion A simultaneous detection method of a variety of shrimp tropomyosins is establised successfully based on ELISA using common peptide epitopes.
标题：基于虾类原肌球蛋白共同表位肽抗体的过敏原检测方法
英文标题：Rapid detection of shrimp allergens based against the polyclonal antibodies of shrimp tropomyosin common peptide epitopes
作者：
刘玉敏 威海出入境检验检疫局
梁占丽 威海出入境检验检疫局
李兆杰 威海出入境检验检疫局
徐成钢 威海出入境检验检疫局
魏玮 威海出入境检验检疫局
张玉春 威海出入境检验检疫局
中文关键词:虾类过敏原,酶联免疫法,共同表位肽,抗体,
英文关键词:shrimp allergen,ELISA,common epitopes peptide,antibody,
发表日期：2017-07-23
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