Food Allergy Research and Resource Program, Department of Food Science and Technology, University of Nebraska, Lincoln, NE 68583-0955, USA.
Biomed Res Int. 2021 Mar 19;2021:6685575. doi: 10.1155/2021/6685575. eCollection 2021.
Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf () and ocean quahog (), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.
海鲜是全球范围内引起食物过敏反应的常见原因。未申报的痕量贻贝可导致敏感个体发生过敏反应。用于测试食品中痕量贻贝存在的工具有限。我们报告了一种夹心 ELISA 的开发，该 ELISA 可用于检测和定量食品中的贻贝蛋白。针对两种商业上重要的贻贝物种（大西洋贻贝（）和美洲帘蛤（））的混合物，用兔和羊制备了抗血清。使用该抗血清构建了夹心 ELISA，并评估了其敏感性和特异性。此外，还分析了用贻贝蛋白加标的模型食品产品，以评估 ELISA 的性能。并与市售的甲壳类动物 ELISA 进行了比较。夹心 ELISA 的定量下限为 2.5 ppm 食品样品中的贻贝蛋白，允许检测可能引发贻贝过敏患者反应的低量贻贝。夹心 ELISA 具有高度特异性，仅对其他软体动物贝类（贻贝和扇贝）表现出交叉反应性。番茄汁和土豆奶油汤中的贻贝蛋白检测良好，回收率分别为 65%至 74%和 74%至 113%。然而，当土豆奶油汤进行高温杀菌时，回收率降至 20%，这可能导致对食品中贻贝含量的低估。市售的甲壳类动物 ELISA 测试不适用于检测贻贝蛋白。本文描述的夹心 ELISA 适用于检测和定量食品中的贻贝蛋白。对于已经高温杀菌的食品产品应格外小心，因为结果可能会被低估。
