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新型食品中产肠毒素金黄色葡萄球菌检测平台。

Novel platform for the detection of Staphylococcus aureus enterotoxin B in foods.

机构信息

U.S. FDA Center for Food Safety and Applied Nutrition, College Park, MD, USA.

出版信息

Appl Environ Microbiol. 2013 Mar;79(5):1422-7. doi: 10.1128/AEM.02743-12. Epub 2012 Dec 14.

Abstract

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.

摘要

食品中葡萄球菌的污染以及由其引发的食源性疾病仍然是全球性的问题。传统方法对食品中金黄色葡萄球菌(金黄色葡萄球菌)和/或肠毒素的筛选既繁琐又费力。如果能从单一食品提取物或培养上清液中,可靠且快速地进行多重检测,将会简化检测过程。本研究建立了一种基于荧光的细胞芯片检测方法,用于检测肠毒素 B(SEB),该方法使用与 T 细胞受体(TCR)的肠毒素特异性 Vβ 结构域(Vβ-TCR)或多克隆抗体偶联的磁性微球。研究表明,Vβ-TCR 与 SEB 的结合亲和力处于皮摩尔范围,与最佳的单克隆抗体相当。偶联的微球经纯化肠毒素验证,并在添加肠毒素的各种食品基质中进行测试。研究表明,Vβ-TCR 或抗体可特异性结合包括牛奶、土豆泥、香草布丁和熟鸡肉在内的四种不同食品基质中的 SEB。传统多克隆抗体和 Vβ-TCR 的使用提供了一个冗余系统,可确保肠毒素的准确鉴定,而标记的微球则允许从单个样品中同时检测多种肠毒素。

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