食品科学 ›› 2024, Vol. 45 ›› Issue (2): 308-314.doi: 10.7506/spkx1002-6630-20230418-179

• 安全检测 • 上一篇    下一篇

基于金纳米花的双信号适配体传感器检测红酒中真菌毒素

祁兴普,董骐玮,朱麟菲,邹婷婷,郑义,张婧怡,李倩   

  1. (1.江苏农牧科技职业学院食品科技学院,江苏 泰州 225300;2.扬州大学食品科学与工程学院,江苏 扬州 225127)
  • 出版日期:2024-01-25 发布日期:2024-02-05
  • 基金资助:
    江苏农牧科技职业学院校科研项目(NSF2021ZR07);泰州市第五期“311工程”科研资助项目(RCPY202115); 泰州市科技支撑计划项目(SSF20210108);江苏省农业科技自主创新项目(CX(21)3005)

A Gold Nanoflower-Based Dual-Signal Fluorescent Aptasensor for Detection of Mycotoxins in Red Wine

QI Xingpu, DONG Qiwei, ZHU Linfei, ZOU Tingting, ZHENG Yi, ZHANG Jingyi, LI Qian   

  1. (1. College of Food Science and Technology, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China; 2. School of Food Science and Engineering, Yangzhou University, Yangzhou 225127, China)
  • Online:2024-01-25 Published:2024-02-05

摘要: 构建基于金纳米花(gold nanoflower,AuFL)和适配体的双信号荧光适配体传感器用于同时检测赭曲霉毒素A(ochratoxin A,OTA)和玉米赤霉烯酮(zearalenone,ZEN)。以AuFL为载体,将含OTA和ZEN适配体的单链DNA共价交联在AuFL表面,进一步将其与两种荧光染料(FAM和Cy3)标记的互补DNA杂交,制备DNA功能化纳米探针。荧光染料和AuFL之间发生荧光共振能量转移(fluorescence resonance energy transfer,FRET)导致纳米探针自身无荧光。加入OTA和ZEN后,FAM和Cy3从探针表面脱落会扰乱FRET过程,导致两者荧光的恢复。优化条件下,该双信号荧光适配体传感器对OTA检测的线性范围为0.05~500 ng/mL,检出限为0.017 ng/mL。对ZEN检测的线性范围为0.1~500 ng/mL,检出限为0.033 ng/mL。该双信号荧光适配体传感器具有良好的选择性和重现性,并能够成功应用于红酒实际样中OTA和ZEN的检测。

关键词: 双信号;适配体传感器;荧光检测;赭曲霉毒素A;玉米赤霉烯酮

Abstract: A dual-signal fluorescent aptasensor based on gold nanoflower (AuFL) was constructed for the simultaneous detection of ochratoxin A (OTA) and zearalenone (ZEN). Single-stranded DNA containing OTA and ZEN aptamers were covalently cross-linked on the surface of AuFL as a nanocarrier and hybridized with complementary DNA labeled with two fluorescent dyes (FAM and Cy3) to prepare a DNA functionalized nanoprobe. Because of fluorescence resonance energy transfer (FRET) between the fluorescent dyes and AuFL, the nanoprobe itself had no fluorescence. When OTA and ZEN were added, FAM and Cy3 were shed from the probe’s surface, disturbing the FRET process and consequently restoring the fluorescence of both fluorescent dyes. Under optimized conditions, the linear range was from 0.05 to 500 ng/mL, and the detection limit was 0.017 ng/mL for OTA; the linear range was from 0.1 to 500 ng/mL, and the detection limit was 0.033 ng/mL for ZEN. The dual-signal fluorescent aptasensor had good selectivity and reproducibility and was successfully applied to the detection of OTA and ZEN in real wine samples.

Key words: dual-signal; aptasensor; ?uorescence detection; ochratoxin A; zearalenone

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