ABSTRACT The optimum conditions of a direct competitive enzyme-linked immunosorbent assay (dcELISA) in regard to different monoclonal antibodies (MAbs), coating conditions, blocking conditions and physicochemical factors of the assay buffer were investigated to develop a broad-specific and sensitive immunoassay for detection of antibacterial synergists (ASGs) in milk, eggs, chicken and pork. Under optimized conditions, a highly broad-specific and sensitive dcELISA for ASGs was obtained based on 5C4 and hapten-C-HRP, demonstrating a 50% specific binding (IC50) for five ASGs of 0.208–9.24 ng mL−1 with cross-reactivity (CR) of 138.2–7.5%. The optimized dcELISA was used to quantify the five ASGs in milk, egg, chicken muscle and beef with the average recoveries of 79.5–105.4% and coefficients of variation (CV) less than 13.4%. The established method with broad specificity and uniform affinity, offered a simple, sensitive, and high-throughput screening tool for the detection of multi- ASGs in animal-derived food.

中文翻译：

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用于筛选多抗菌增效剂的高度广泛特异性和灵敏的直接竞争性酶联免疫吸附试验：试验优化和在动物源性食品中的应用

摘要 研究了直接竞争性酶联免疫吸附试验 (dcELISA) 在不同单克隆抗体 (MAb) 方面的最佳条件、包被条件、封闭条件和测定缓冲液的理化因素，以开发一种广泛特异性和灵敏的免疫测定法。检测牛奶、鸡蛋、鸡肉和猪肉中的抗菌增效剂 (ASGs)。在优化条件下，基于 5C4 和半抗原-C-HRP 获得了针对 ASG 的高度广泛特异性和灵敏的 dcELISA，证明对 0.208-9.24 ng mL-1 的五种 ASG 具有 50% 的特异性结合 (IC50)，具有交叉反应性(CR) 为 138.2–7.5%。优化的 dcELISA 用于量化牛奶、鸡蛋、鸡肌肉和牛肉中的五种 ASG，平均回收率为 79.5-105.4%，变异系数 (CV) 小于 13.4%。