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基于适配体和金纳米棒的伏马菌素 B1 荧光和 SERS 分析检测法。

Aptamer and gold nanorod-based fumonisin B1 assay using both fluorometry and SERS.

机构信息

State Key Laboratory of Biobased Material and Green Papermaking, College of Food Science and Engineering, Qilu University of Technology, Shandong Academy of Sciences, Jinan, 250353, China.

State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100094, China.

出版信息

Mikrochim Acta. 2020 Mar 11;187(4):215. doi: 10.1007/s00604-020-4192-0.

Abstract

An aptamer-based assay is presented for the determination of fumonisin B1 (FB1). It is bimodal in that both surface-enhanced Raman spectroscopy (SERS) and fluorometry are applied for quantitation. It makes use of platinum-coated gold nanorod (AuNR) and DNA sequences. The complementary DNA of aptamer (cDNA) against FB1 is immobilized on the surface of AuNR. The aptamer of FB1 modified with Cy5.5 are complementarily hybridized with cDNA. In the absence of FB1, the aptamer and its cDNA associate. In this situation, strong SERS and weak fluorescence signals are obtained. In the presence of FB1, the aptamer disassociates with its cDNA and binds the target. As the concentration of FB1 increases, the SERS and fluorescence signal intensities of the mixture are gradually decreased and increased, respectively. Under optimized conditions, the SERS signal at 1366 cm decreases linearly in the 10-500 pg mL concentration range with the calibration equation of y = 1997lgx-594 (the coefficient of determination is 0.998). The fluorescence signal at 670 nm increases linearly in the 10-250 pg mL concentration range with the calibration equation of y = 500lgx-383 (the coefficient of determination is 0.991). The assay was applied to the determination of FB1 contents in spiked corn samples. The average recoveries ranged from 92 to 107%, confirming the practicality of this method. The results obtained by this assay are in good agreement with that of LC-MS/MS method. Graphical abstractSchematic illustration of a bimodal aptasensor based on surface enhanced Raman scattering (SERS) and fluorescence change for the detection of fumonisin B1 (FB1).

摘要

基于适配体的测定法用于检测伏马菌素 B1(FB1)。它是双模态的,因为同时应用了表面增强拉曼光谱(SERS)和荧光法进行定量。它利用了铂涂层金纳米棒(AuNR)和 DNA 序列。针对 FB1 的适配体互补 DNA(cDNA)固定在 AuNR 的表面。经 Cy5.5 修饰的 FB1 适配体与 cDNA 互补杂交。在没有 FB1 的情况下,适配体与其 cDNA 结合。在这种情况下,会得到强的 SERS 和弱的荧光信号。在存在 FB1 的情况下,适配体与 cDNA 解离并与靶标结合。随着 FB1 浓度的增加,混合物的 SERS 和荧光信号强度逐渐降低和增加。在优化条件下,1366 cm 处的 SERS 信号在 10-500 pg mL 浓度范围内呈线性降低,校准方程为 y = 1997lgx-594(决定系数为 0.998)。670nm 处的荧光信号在 10-250 pg mL 浓度范围内呈线性增加,校准方程为 y = 500lgx-383(决定系数为 0.991)。该测定法用于测定加标玉米样品中的 FB1 含量。平均回收率在 92%至 107%之间,证实了该方法的实用性。该测定法的结果与 LC-MS/MS 方法的结果吻合良好。

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