化学发光光纤免疫传感器的研制及其对谷物中呕吐毒素的快速检测
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作者单位:

华中农业大学食品科学技术学院/环境食品学教育部重点实验室,武汉 430070

作者简介:

黄伟,E-mail:2020309120024@webmail.hzau.edu.cn

通讯作者:

陈翊平,E-mail:chenyiping@mail.hzau.edu.cn

中图分类号:

TS207.5

基金项目:

岭南现代农业重点实验室项目(NZ2021036);国家重点研发计划项目(2019YFC1605502)


Development of a chemiluminescent optical fiber immunosensor and its application in rapid detection of deoxynivalenol in grains
Author:
Affiliation:

College of Food Science and Technology,Huazhong Agricultural University/ Key Laboratory of Environment Correlative Dietology,Ministry of Education,Wuhan 430070,China

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    摘要:

    为实现谷物中呕吐毒素(deoxynivalenol,DON)的快速、灵敏检测,采用生物传感和化学发光技术相结合的方式,研制应用于现场快速检测谷物中呕吐毒素的化学发光光纤免疫传感器,并搭建用于化学发光信号检测的便携式传感平台。整个检测平台集成到一个32 cm×15 cm×25 cm的黑色避光盒中,质量小于3 kg。结果显示:所研制的化学发光光纤免疫传感器在0.1~1 200 ng/mL呈现出良好的线性关系,检出限为62.7 pg/mL;功能化的光纤探针能够稳定保存30 d且具有良好的特异性和抗干扰性;其应用于谷物样品的呕吐毒素检测回收率为81.00%~107.33%,相对标准偏差小于8.69%。结果表明,该方法较ELISA法的灵敏度提高了30倍,线性范围拓宽了3个数量级,适合应用于谷物样品中呕吐毒素的现场快速检测。

    Abstract:

    Deoxynivalenol (DON),as a trichothecenes metabolite produced by Fusarium,widely exists in grains including wheat,rice and corn,which seriously threatens human health.Therefore,based on the theoretical guidance of biochemistry,immunology and materials science,we constructed a chemiluminescent fiber optic immunosensor for the on-site rapid detection of DON in grains by using a combination of biosensing recognition and chemiluminescence technology.At the same time,a portable sensing platform for detecting the chemiluminescence signal was integrated into a black light-proof cassette of 32 cm×15 cm× 25 cm,and the weight was less than 3 kg.The results showed that Fourier transform infrared, energy dispersive spectrometry and atomic force microscopy confirmed the successful preparation of fully antigenicized fiber probes. In addition, the constructed chemiluminescence optical fiber immunosensor had a good linear relationship between 0.1-1 200 ng/mL,with the detection limit of 62.7 pg/mL.The functionalized optical fiber probe can be stably stored for 30 days and has good specificity and anti-interference.The recovery rate of DON applied to grain samples was 81.00%-107.33%,and the relative standard deviation was less than 8.69%.Compared with ELISA,the sensitivity of this method was increased by 30 times,and the linear range was broadened by 3 orders of magnitude,indicating that it is suitable for the on-site rapid detection of DON in grain samples.

    表 1 化学发光光纤免疫传感器与参考文献中呕吐毒素检测方法的对比Table 1 Comparison of chemiluminescent fiber optic immunosensors and DON detection methods in reference
    表 2 加标谷物中检测呕吐毒素的回收率Table 2 Recovery of DON in spiked grains
    图1 化学发光光纤免疫传感器检测呕吐毒素原理图Fig.1 Schematic diagram of chemiluminescent fiber optic immunosensor for detection of DON
    图2 化学发光光纤免疫传感器检测平台的结构Fig.2 Structure of the detection platform for chemiluminescent fiber optic immunosensor
    图3 傅里叶变换红外光谱图(A)、能量色散光谱图(B)、修饰前的光纤探针(C)和修饰完全抗原(BSA-DON)后的光纤探针(D)的原子力显微镜图Fig.3 Fourier transform infrared spectrum (FT-IR) characterization diagram(A),energy dispersive spectroscopy diagram(B),atomic force microscope characterization diagram of the optical fiber probe without modification (C) and after BSA-DON modification (D)
    图4 不同时间下光纤探针的稳定性Fig.4 Stability of fiber optic probe at different time
    图5 完全抗原、呕吐毒素抗体和免疫反应时间对化学发光强度的影响Fig.5 Effects of complete antigen,DON-antibody and immune response time on chemiluminescence intensity
    图6 化学发光光纤免疫传感器检测DON的特异性(A)和抗干扰性能(B)Fig.6 Specificity analysis (A) and anti-interference performance analysis (B) of chemiluminescent fiber optic immunosensor for DON detection
    图7 化学发光光纤免疫传感器检测DON的标准曲线(A)和线性范围(B)Fig.7 Standard curve (A) and linear range (B) of chemiluminescent fiber optic immunosensor for DON detection
    图8 酶联免疫吸附法(ELISA)检测DON的标准曲线(A)和线性范围(B)Fig.8 Standard curve (A) and linear range (B) of enzyme-linked immunosorbent assay for DON detection
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黄伟,何慧禹,鲁鹏,陈翊平.化学发光光纤免疫传感器的研制及其对谷物中呕吐毒素的快速检测[J].华中农业大学学报,2022,41(5):273-282

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  • 收稿日期:2022-05-03
  • 在线发布日期: 2022-10-11
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