利用邻位连接技术检测花生过敏原成分  

A Proximity Ligation Assay of Peanut Allergen

作　　者：张霞[1] 赵良娟[1] 高琳[1] 温华蔚 王飞雪[2] 董焱[2] 刘掘茫 刘寅[2] 郑文杰[1] 

机构地区：[1]天津出入境检验检疫局,天津300461 [2]南开大学,天津300071

出　　处：《食品研究与开发》2016年第9期148-151,共4页Food Research and Development

基　　金：质检总局科研专项<邻位连接技术高效检测食品中过敏原蛋白方法的研究>资助(2014IK093)

摘　　要：建立一种新型花生过敏原成分的快速筛选检测方法--邻位连接技术检测方法。针对花生过敏原选择适当的鼠源单克隆抗体,并合成针对花生过敏原的兔源多克隆抗体然后与磁颗粒偶联,而后将羊抗兔二抗与一条DNA分子耦联制成亲和探针1,将另一条寡核苷酸序列与羊抗鼠二抗耦联制成亲和探针2,上述成分能够与一连结DNA组合成邻位连接复合物,建立邻位连接检测花生过敏原的方法。使用3种花生过敏原标准品和10种确定不含花生过敏原的样本进行特异性试验;通过稀释花生过敏原标准品进行灵敏度验证。建立方法具有很好的特异性;检测标准品的灵敏度为0.01 ng/m L,比普通ELISA试剂盒灵敏度提高了1 000倍。建立的邻位连接检测花生过敏原的方法能够检测出使用目前常规方法不能检测到的花生过敏原,具有重要的实际意义。To establish a new method with high-sensitivity for detection of peanut allergen-proximity ligation assay. The proximity ligation assay compound consists of monoclonal antibody from mouse to peanut allergen, polyclonal antibody from rabbit to peanut allergen, secondary antibody goat anti-mouse, secondary antibody goat anti-rabbit, DNA nucleotide probe 1 and 2, DNA connector. Three kinds of peanut allergen reference standards and ten samples without peanut allergen including hazelnut and apricot seed references were evaluated the specificity of the proximity ligation assay. The dilution of the peanut allergen reference standards were used to evaluate the sensitivity of the assay. The assay established in present study was highly specific and sensitive. The limit of detection was 0.01 ng/mL for the peanut allergen reference standards, which was 1 000 times better than the ELISA method. The method can detect the invisible peanut allergen objects using traditional methods. It is very useful for detection of peanut allergen in the actual detection work.

关 键 词：花生过敏原 邻位连接 亲和探针 磁颗粒 

分 类 号：TS207.3[轻工技术与工程—食品科学]