Department of Physical and Analytical Chemistry & Institute of Biotechnology of Asturias, University of Oviedo, c/Julián Clavería 8, 33006, Oviedo, Spain.
Department of Chemical and Environmental Engineering & Institute of Biotechnology of Asturias, University of Oviedo, Oviedo, Spain.
Mikrochim Acta. 2023 Jun 19;190(7):264. doi: 10.1007/s00604-023-05834-8.
The aim of this study was to develop a sensitive lateral flow immunoassay (LFIA) for the rapid detection of Escherichia coli (E. coli) O157:H7, a pathogen contributor to diseases and fatalities worldwide. Au nanoparticles with high stability, uniform size, and shape were synthesized and coated with heterobifunctional PEG polymer with carboxyl groups, and they were bioconjugated to be used as label in sandwich-LFIA. Then, a silver enhancement strategy was developed as an accessible, rapid, and cost-effective approach for signal amplification to reduce the limit of detection (LOD). The optimal results were achieved when a solution of silver nitrate and hydroquinone/citrate buffer was added to the strips for 4 min. This led to a decrease in the visual LOD from 2 × 10 (CFU mL) to 2 × 10 (CFU mL), resulting in a threefold improvement in sensitivity compared to the conventional LFIA system. The specificity of the system was evaluated by using non-target bacteria (E. coli BL21 and E. coli T515) and its reliability was determined by testing commercial food samples (milk, tap water, and orange juice), demonstrating its effectiveness for quickly detecting pathogenic bacteria in food products.
本研究旨在开发一种灵敏的侧向流动免疫分析（LFIA）方法，用于快速检测大肠杆菌（E. coli）O157:H7，这种病原体是全球疾病和死亡的致病因素。高稳定性、均一尺寸和形状的金纳米颗粒被合成，并涂覆具有羧基的杂双功能 PEG 聚合物，然后被生物偶联用作夹心 LFIA 的标记。随后，开发了一种银增强策略，作为一种可及、快速且具有成本效益的信号放大方法，以降低检测限（LOD）。当将硝酸银和对苯二酚/柠檬酸盐缓冲液的溶液添加到条带中 4 分钟时，可获得最佳结果。这导致视觉 LOD 从 2×10（CFU mL）降低至 2×10（CFU mL），与传统 LFIA 系统相比，灵敏度提高了三倍。通过使用非靶标细菌（大肠杆菌 BL21 和大肠杆菌 T515）评估了该系统的特异性，并通过测试商业食品样品（牛奶、自来水和橙汁）确定了其可靠性，证明了其在快速检测食品中致病菌方面的有效性。
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