肠毒素
假阳性悖论
免疫分析
金黄色葡萄球菌
微生物学
检出限
抗体
毒力
化学
计算生物学
生物
色谱法
细菌
计算机科学
免疫学
大肠杆菌
生物化学
遗传学
机器学习
基因
作者
Yan Chen,Xiatong Wang,Haofen Wu,Xiaoling Zhang,Yingming Xu,Gege Yu,Xiaojing Liu,Qin Yao,Jianlong Wang,Yanwei Ji
出处
期刊:Food Control
[Elsevier]
日期:2024-06-01
卷期号:160: 110313-110313
被引量:2
标识
DOI:10.1016/j.foodcont.2024.110313
摘要
Staphylococcal enterotoxin A (SEA), a virulence factor produced by Staphylococcus aureus (S. aureus), poses a serious threat to human health. The rapid and sensitive detection of SEA is crucial for preventing the spread of contaminated food and safeguarding public health. The conventional enzyme-linked immunosorbent assay (ELISA) has encountered limitations in detecting SEA due to the false-positives caused by the combination of the Fc-terminus of the antibodies and staphylococcal protein A (SpA) and its insufficient sensitivity that fails to meet current detection requirements. Therefore, nanobodies (Nbs) without Fc-termini can replace traditional antibodies to prevent false-positives. We developed a "one to two" sandwich ELISA based on identical capture Nbs and double detection phage-displayed Nbs (OtTNb ELISA), which achieved detection with an LOD of 0.43 ng/mL in a linear range of 0.5–512 ng/mL. The sensitivity was over 3.4 times higher than that of the sandwich ELISA based on mAbs, and the linear range exceeded over 8 times. The OtTNb ELISA was successfully applied to the cross-reactivity test and the actual recovery of the sample. This study provides a sensitive tool for detecting staphylococcal enterotoxins (SEs) in food and offers insights into developing detection methods for various foodborne pathogens and toxins.
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