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Competitive HRP-Linked Colorimetric Aptasensor for the Detection of Fumonisin B1 in Food based on Dual Biotin-Streptavidin Interaction

机译:基于双重生物素-链霉亲和素相互作用的竞争性HRP链接比色适体传感器检测食品中的伏马菌素B1

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摘要

Fumonisin B1 (FB1) is the most prevalent and toxic form among fumonisin homologues which are produced by fusarium species and it contaminates various types of food products, posing serious health hazards for humans and animals. In this work, a colorimetric assay for the detection of FB1 has been developed based on competitive horseradish peroxidase (HRP)-linked aptamer and dual biotin-streptavidin interaction. In short, a biotinylated aptamer of FB1 was immobilized on the microplate by biotin-streptavidin binding; the complementary strand (csDNA) of the aptamer was ligated with HRP by biotin-streptavidin binding again to form a csDNA-HRP sensing probe, competing with FB1 to bind to the aptamer. The color change can be observed after the addition of chromogenic and stop solution, thereby realizing the visual detection of FB1. Under optimal conditions, good linearity was observed within the concentration range of 0.5 to 300 ng/mL, with a detection of limit of 0.3 ng/mL. This assay is further validated by spike recovery tests towards beer and corn samples, it provides a simple, sensitive and reliable method for the screening of FB1 in food samples and may be potentially used as an alternative to conventional assays.
机译:伏马菌素B1(FB1)是镰刀菌属物种产生的伏马菌素同系物中最普遍和最具毒性的形式,它污染了各种食品,对人类和动物构成了严重的健康危害。在这项工作中,基于竞争性辣根过氧化物酶(HRP)连接的适体和双重生物素-链霉亲和素相互作用,开发了用于检测FB1的比色测定法。简而言之,通过生物素-链霉亲和素结合将FB1的生物素化适体固定在微板上。通过再次结合生物素-链霉亲和素,将适体的互补链(csDNA)与HRP连接,以形成csDNA-HRP传感探针,与FB1竞争与适体结合。加入生色和终止溶液后可以观察到颜色变化,从而实现了FB1的视觉检测。在最佳条件下,在0.5至300 ng / mL的浓度范围内观察到良好的线性,检出限为0.3 ng / mL。通过对啤酒和玉米样品的加标回收率测试进一步验证了该分析方法,它为食品样品中FB1的筛选提供了一种简单,灵敏和可靠的方法,并且有可能被用作常规分析方法的替代方法。

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