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Novel Platform for the Detection of Staphylococcus aureus Enterotoxin B in Foods

机译:检测食品中金黄色葡萄球菌肠毒素B的新型平台

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Staphylococcal contamination of food products and staphylococcal food-borne illnesses continue to be a problem worldwide. Screening of food for the presence of Staphylococcus aureus and/or enterotoxins using traditional methods is laborious. Reliable and rapid multiplex detection methods from a single food extract or culture supernatant would simplify testing. A fluorescence-based cytometric bead array was developed for the detection of staphylococcal enterotoxin B (SEB), using magnetic microspheres coupled with either an engineered, enterotoxin-specific Vβ domain of the T-cell receptor (Vβ-TCR) or polyclonal antibodies. The binding affinity of the Vβ-TCR for SEB has been shown to be in the picomolar range, comparable to the best monoclonal antibodies. The coupled beads were validated with purified enterotoxins and tested in a variety of food matrices spiked with enterotoxins. The Vβ-TCR or antibody was shown to specifically bind SEB in four different food matrices, including milk, mashed potatoes, vanilla pudding, and cooked chicken. The use of traditional polyclonal antibodies and Vβ-TCR provides a redundant system that ensures accurate identification of the enterotoxin, and the use of labeled microspheres permits simultaneous testing of multiple enterotoxins from a single sample.
机译:食品的葡萄球菌污染和食源性葡萄球菌仍然是世界范围的问题。使用传统方法筛查食物中是否存在金黄色葡萄球菌和/或肠毒素是费力的。从单一食品提取物或培养物上清液中可靠而快速的多重检测方法将简化测试。使用磁性微球与T细胞受体(Vβ-TCR)的工程化肠毒素特异性Vβ结构域或多克隆抗体偶联,开发了基于荧光的细胞计数珠阵列,用于检测葡萄球菌肠毒素B(SEB)。 Vβ-TCR对SEB的结合亲和力已显示在皮摩尔范围内,与最佳单克隆抗体相当。用纯化的肠毒素验证偶联的珠子,并在掺有肠毒素的各种食品基质中进行测试。已显示Vβ-TCR或抗体在四种不同的食品基质(包括牛奶,土豆泥,香草布丁和煮熟的鸡肉)中与SEB特异性结合。传统多克隆抗体和Vβ-TCR的使用提供了一个冗余系统,可确保准确识别肠毒素,而标记微球的使用可同时检测单个样品中的多种肠毒素。

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