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An aptamer-based fluorescence probe for facile detection of lipopolysaccharide in drinks

机译:基于适体的荧光探针,可轻松检测饮料中的脂多糖

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Bacterial toxin contamination is a serious food safety issue. Rapid, facile detection of bacterial toxins in food is crucial to control their harmful effects on human health. In this study, a facile, selective, and cost-effective method with a 6-carboxy-X-rhodamine labeled lipopolysaccharide binding aptamer (ROX-LBA)/GO fluorescent probe was developed based on the specific recognition characteristics of aptamers and the super fluorescence quenching efficiency of graphene oxide (GO) and used for the detection of lipopolysaccharides (LPS). ROX-LBA is adsorbed on the GO surface because of its strong π–π stacking interactions in the flexible single-stranded state, and fluorescence is quenched by GO via F?rster resonance energy transfer (FRET). After adding LPS, the aptamer binds to LPS and forms a LPS/ROX-LBA complex, which exhibits weak binding affinity with GO. ROX-LBA cannot effectively combine with GO, and fluorescence is restored. The concentration of LPS can be quantitatively analyzed by observing fluorescence changes in the sensing system. The fluorescence intensity proportionally increases with LPS concentration (25–1600 ng mL?1), and the detection limit is 15.7 ng mL?1. Hence, the proposed method provides an alternative strategy for the specific detection of endotoxin in drink samples (e.g., apple juice and beer) and could be widely applied to quantify various bacterial toxins by replacing the aptamer sequences.
机译:细菌毒素污染是一个严重的食品安全问题。快速,简便地检测食物中的细菌毒素对于控制其对人体健康的有害影响至关重要。在这项研究中,基于适体的特异性识别特性和超荧光性,开发了一种简便,选择性且经济高效的方法,该方法使用6-羧基-X-罗丹明标记的脂多糖结合适体(ROX-LBA)/ GO荧光探针氧化石墨烯(GO)的猝灭效率,用于检测脂多糖(LPS)。 ROX-LBA由于在柔性单链状态下强烈的π-π堆积相互作用而被吸附在GO表面,并且通过F 共振能量转移(FRET)通过GO 猝灭了荧光。加入LPS后,适体与LPS结合并形成LPS / ROX-LBA复合物,该复合物显示出与GO的弱结合亲和力。 ROX-LBA无法与GO有效结合,并且荧光得以恢复。 LPS的浓度可以通过观察传感系统中的荧光变化进行定量分析。荧光强度随LPS浓度(25–1600 ng mL ?1 )成比例增加,检出限为15.7 ng mL ?1 。因此,所提出的方法为饮料样品(例如,苹果汁和啤酒)中内毒素的特异性检测提供了另一种策略,并且可以通过替换适体序列而广泛应用于定量各种细菌毒素。

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