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基于抗生素亲和策略和荧光猝灭效应的李斯特菌双重识别夹心荧光法。

Sandwich fluorometric method for dual-role recognition of Listeria monocytogenes based on antibiotic-affinity strategy and fluorescence quenching effect.

机构信息

College of Food Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China.

College of Food Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China.

出版信息

Anal Chim Acta. 2022 Aug 15;1221:340085. doi: 10.1016/j.aca.2022.340085. Epub 2022 Jun 14.

Abstract

In this work, a sandwich fluorometric method for dual-role recognition of L. monocytogenes was developed based on antibiotic-affinity strategy and fluorescence quenching effect for sensitive and rapid detection of L. monocytogenes in ham samples. Vancomycin (Van) was conjugated with magnetic nanoparticles (MNPs) to recognize and capture target bacteria. Biotinylated aptamers were used to bind specifically to L. monocytogenes through the cell wall. The two agents recognized target bacteria at different binding sites showing satisfied specificity. The upconversion fluorescence response signal could be enlarged by using the inner filter effect (IFE) between the colored products produced by enzyme-catalyzing substrate and upconversion nanoparticles (UCNPs). The change in fluorescence intensity could represent the concentration of target bacteria over 10-2 × 10 CFU mL. The developed sandwich fluorimetric method achieved a low detection limit (LOD) of 2.8 × 10 CFU mL. Overall, the constructed fluorometric sensor could provide a simple and reliable method for the detection of L. monocytogenes.

摘要

本工作基于抗生素亲和策略和荧光猝灭效应,开发了一种用于李斯特菌双重识别的三明治荧光法,可用于灵敏、快速地检测火腿样品中的李斯特菌。万古霉素(Van)与磁性纳米粒子(MNPs)偶联,以识别和捕获靶细菌。生物素化适体通过细胞壁特异性结合李斯特菌。这两种试剂在不同的结合位点识别靶细菌,表现出满意的特异性。上转换荧光响应信号可以通过酶催化底物和上转换纳米粒子(UCNPs)产生的有色产物之间的内滤效应(IFE)放大。荧光强度的变化可以代表目标细菌的浓度超过 10-2×10 CFU mL-1。所开发的三明治荧光法的检测限(LOD)低至 2.8×10 CFU mL-1。总的来说,所构建的荧光传感器可以为李斯特菌的检测提供一种简单可靠的方法。

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