黄曲霉毒素B1荧光/LSPR双信号适配体传感器的构建及应用

doi:10.3964/j.issn.1000-0593(2025)04-1123-06

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摘要：食源性感染可能会引起新疾病产生和原有传染病再次出现，是发达国家和发展中国家疾病和死亡的主要原因之一，严重影响人类的公共卫生安全。黄曲霉毒素B1(Aflatoxin B1, AFB1)是食源性霉菌毒素中毒性最大的一种，它可能通过受污染的谷类食品对人体和动物健康造成严重危害。AFB1在食品中的普遍污染，加之人们对食品安全的日益重视，决定AFB1快速准确的检测方法发展具有越来越重要的意义。光学生物传感器将生物特异性识别元件和生物材料组合起来，将生物反应转化为可测量的信号，由于检测快速、灵敏度高、前处理简单等优点，光学生物传感器在生物检测传感领域有着广阔应用前景。本文利用金纳米球粒子(AuNPs)的局域表面等离子共振（LSPR）特性和荧光抑制性，构建以适配体(Aptamer, Apt)保护的金纳米粒子为信号探针的LSPR/荧光双信号输出适配体传感器，用于AFB1的定量检测。Apt保护的AuNPs在荧光分子罗丹明异硫氰酸酯（RBITC）存在的情况下分散良好，金纳米粒子的LSPR性能稳定，且RBITC荧光强度明显；由于AFB1与Apt之间的特异性，在目标物AFB1竞争作用下，AuNPs失去Apt保护，迅速被阳离子溶液RBITC分子吸附，造成金纳米粒子团聚，溶液颜色和LSPR性能发生改变，且RBITC分子的荧光基团在金纳米粒子作用下发生荧光猝灭；随后，在铁氰化钾(K₃[Fe(CN)₆])和碘化钾(KI)组成的刻蚀剂溶液作用下，金纳米粒子被氧化刻蚀消失，RBITC分子从金纳米粒子上脱落，荧光性能得到恢复，由此建立AFB1浓度与AuNPs的LSPR性能改变前后吸光度比值及RBITC分子荧光强度之间的定量关系。构建的双信号输出适配体传感器能够在0.000 1~1 ng·mL^-1浓度范围内实现AFB1的定量检测，在真实样品检测中，LSPR实验回收率可达95.6%~105%，荧光光谱检测回收率为92.3%~118%。本文将纳米技术、生物技术、适配体技术、霉菌毒素检测技术结合起来构建的基于LSPR/荧光双信号适配体传感器是一种很有潜力且有望用于AFB1现场快速检测的技术。

关键词：荧光光谱；局域表面等离子共振；黄曲霉毒素B1；金纳米粒子；罗丹明异硫氰酸酯

Abstract：Foodborne infections pose a substantial hazard to public health worldwide because they play a significant role in establishing and reemerging infectious illnesses. Aflatoxin B1 (AFB1) is a very toxic mycotoxin mainly present in contaminated cereal and poses a risk to human and animal health. With the widespread contamination of AFB1 in food and people's increasing attention to food safety, a quick, accurate, and trustworthy technique for detecting AFB1 in food products is desperately needed. Optical biosensors combine biological-specific recognition elements and biological materials to convert biological reactions into measurable signals. Due to the advantages of fast detection, high sensitivity, and simple pre-processing, optical biosensors have broad application prospects in biological detection sensing. Therefore, in this work, we designed a dual-signal output aptamer sensor for quantitatively detecting AFB1 based on the fluorescence quenching capabilities of gold nanoparticles (AuNPs) and Localized Surface Plasmon Resonance (LSPR). The sensor design used auNPs shielded by AFB1-specific aptamers as signal probes. The competitive binding of AFB1 was subjected to cause the aptamers to separate from the AuNPs, allowing the exposed AuNPs to adsorb Rhodamine B Isothiocyanate (RBITC) quickly. Furthermore, the AuNPs aggregation and quenching of the fluorescence of RBITC were exploited to measure the optical index. The fluorescence of RBITC was restored by further oxidation and etching of the AuNPs using potassium ferricyanide (K[Fe(CN)₆]) and potassium iodide (KI) solution, allowing for precise quantification of AFB1. The results revealed high precision as the developed sensor exhibited a wide detection range for AFB1, ranging from 0.000 1 to 1 ng·mL^-1. During actual sample testing, recovery rates from the LSPR-based approach ranged from 95.6% to 105%, whereas recovery rates from fluorescence-based detection were between 92.3% and 118%. Using this novel approach for mycotoxin detection, the LSPR/fluorescence dual-signal aptamer sensor holds tremendous potential for the quick and on-site detection of AFB1, providing a useful instrument to improve food safety.

Key words：Fluorescence spectrum; LSPR; AFB1; AuNPs; Rhodamine isocyanate

收稿日期: 2024-03-27 修订日期: 2024-09-13

中图分类号:

Q03

基金资助: 河南省重点研发项目(221111320700)，国家自然科学基金项目(31671581)，国家重点研发计划项目(2017YFD0801204)，海关总署项目(2023HK128), 2025年河南省科技攻关项目（252102320259）资助

通讯作者: 张燕燕 E-mail: zyanyan0923@163.com

作者简介: 朱明明，1981年生, 郑州海关技术中心硕士研究生 e-mail: zhumingming@customs.gov.cn

引用本文:

朱明明, 胡建东, 张守杰, 李光辉, 张燕燕. 黄曲霉毒素B1荧光/LSPR双信号适配体传感器的构建及应用[J]. 光谱学与光谱分析, 2025, 45(04): 1123-1128.
ZHU Ming-ming, HU Jian-dong, ZHANG Shou-jie, LI Guang-hui, ZHANG Yan-yan. Construction and Application of Fluorescence/LSPR Dual Signal Aptamer Sensor for Aflatoxin B1 Detection. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2025, 45(04): 1123-1128.

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[1] Marchese S, Polo A, Ariano A, et al. Toxins, 2018, 10(6): 214.
[2] Rushing B R, Selim M I. Food and Chemical Toxicology, 2019, 124: 81.
[3] Benkerroum N. International Journal of Environmental Research and Public Health, 2020, 17(2): 423.
[4] Li M, Lu W Y, Mao Y H, et al. Talanta, 2023, 251: 123798.
[5] Ghali R, Belouaer I, Hdiri S, et al. Journal of Food Composition and Analysis, 2009, 22(7-8): 751.
[6] Dai C S, Sharma G, Liu G Y, et al. Evironmental Pollution, 2024, 345: 123474.
[7] Nolan P, Auer S, Spehar A, et al. Food Additives and Contaminants Part A: Chemistry Analysis Control Exposure & Risk Assessment, 2019, 36(5): 800.
[8] Yin S P, Niu L Q, Liu Y F. Molecules, 2022, 27(19): 6141.
[9] Bhardwaj H, Rajesh, Sumana G. Journal of Food Science and Technology, 2022, 59(1): 12.
[10] Zhang Y, Chen X F, Xie X Y, et al. Current Analytical Chemistry, 2024, 20(4): 242.
[11] Yadav N, Yadav S S, Chhillar A K, et al. Food and Chemical Toxicology, 2021, 152: 112201.
[12] CHEN Peng, LU Feng, ZHAO Yun-li(陈鹏，陆峰，赵云丽). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2024, 44(2): 413.
[13] Tang Y J, Cui S Q, Pu S Z. Journal of Fluorescence, 2016, 26(4): 1421.
[14] Pereira-Barros M A, Daeid N N, Adegoke O. Journal of Photochemistry and Photobiology A: Chemistry, 2021, 418: 113384.
[15] Manju S, Sreenivasan K. Talanta, 2011, 85(5): 2643.
[16] Amina S J, Guo B A. International Journal of Nanomedicine, 2020, 15: 9823.
[17] He H R，Sun D W，Pu H B, et al. Food Chemistry, 2020, 324: 126832.
[18] Deng Y, Chen Y R, Zhou X D. Acta Chimica Slovenica, 2018, 65(2): 271.
[19] Gao S J, Li Z, Sun Z C, et al. Chinese Journal of Polymer Science, 2020, 38(6): 587.
[20] Kalipillai P, Mani E. Physical Chemistry Chemical Physics, 2021, 23(34): 18618.
[21] Li Y, Hu Y J. Applied Physis Letters, 2013, 102(13): 133103.