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Detection of staphylococcal enterotoxins by enzyme-linked immunosorbent assays and radioimmunoassays: comparison of monoclonal and polyclonal antibody systems.

机译:通过酶联免疫吸附测定和放射免疫测定法检测葡萄球菌肠毒素:单克隆和多克隆抗体系统的比较。

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Murine monoclonal antibodies reactive with at least one of the serological types of staphylococcal enterotoxin were examined for use in assay systems for the detection of enterotoxin at the level of 1.0 ng of enterotoxin per ml. An antibody sandwich enzyme-linked immunosorbent assay was devised for each toxin type by identifying an effective antibody pair. One antibody (the coating antibody) was coated onto a polystyrene plate and removed the enterotoxin from the test solution; the second antibody (the probing antibody) was conjugated to horseradish peroxidase and detected the captured toxin. Enterotoxins A and E could be detected in the same system by the use of cross-reacting monoclonal antibodies. All subtypes of enterotoxin C could be detected in one assay system. Two effective systems were described for each of types B and D. Each of these systems, when compared with the homologous enterotoxin-specific polyclonal rabbit antibody systems, was found to compare favorably. The monoclonal enzyme-linked immunosorbent assay systems for the detection of enterotoxins A and C2 were examined for a variety of food extracts; no abnormal interference could be detected from these extracts. The monoclonal antibody systems were also compared with the homologous enterotoxin-specific polyclonal serum for the detection of enterotoxin by the competitive radioimmunoassay (RIA). Single monoclonal antibodies generally did not perform as well in the RIA as did the homologous toxin-specific polyclonal serum. However, pools of monoclonal antibodies were prepared that approached the sensitivity and precision of the polyclonal system for the detection of each toxin by the RIA.
机译:检查了与至少一种血清型葡萄球菌肠毒素有反应性的鼠单克隆抗体,用于检测系统中检测肠毒素的水平为每毫升1.0 ng肠毒素。通过鉴定有效的抗体对,针对每种毒素设计了抗体三明治酶联免疫吸附测定。将一种抗体(包被抗体)包被在聚苯乙烯板上,并从测试溶液中去除肠毒素;第二种抗体(探测抗体)与辣根过氧化物酶偶联,并检测到捕获的毒素。通过使用交叉反应单克隆抗体,可以在同一系统中检测到肠毒素A和E。肠毒素C的所有亚型都可以在一个测定系统中检测到。对于B型和D型,分别描述了两个有效的系统。与同源肠毒素特异性多克隆兔抗体系统相比,这些系统中的每一个都具有良好的比较性。检查了用于检测肠毒素A和C2的单克隆酶联免疫吸附测定系统的各种食品提取物。从这些提取物中未检测到异常干扰。还将单克隆抗体系统与同源肠毒素特异性多克隆血清进行了比较,以通过竞争性放射免疫测定(RIA)检测肠毒素。单一单克隆抗体通常在RIA中的表现不如同源毒素特异性多克隆血清。但是,制备的单克隆抗体库接近用于RIA检测每种毒素的多克隆系统的灵敏度和精确度。

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