食品科学 ›› 2024, Vol. 45 ›› Issue (19): 227-234.doi: 10.7506/spkx1002-6630-20240226-136

• 安全检测 • 上一篇    下一篇

碳化钒免疫层析法检测玉米中T-2毒素

罗昌伟, 陈晓阳, 黄现青, 宋莲军, 张西亚, 补彤   

  1. (河南农业大学食品科学技术学院,河南 郑州 450002)
  • 出版日期:2024-10-15 发布日期:2024-10-12
  • 基金资助:
    国家自然科学基金青年科学基金项目(32302208);中国博士后科学基金资助项目(2023M741056); 2022年河南省研究生联合培养基地项目(YJS2022JD16);河南省高校科技创新团队支持计划资助项目(23IRTSTHN023)

Development of Vanadium Carbide-Based Immunochromatographic Assay for T-2 Toxin Detection in Maize Samples

LUO Changwei, CHEN Xiaoyang, HUANG Xianqing, SONG Lianjun, ZHANG Xiya, BU Tong   

  1. (School of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China)
  • Online:2024-10-15 Published:2024-10-12

摘要: 旨在构建一种碳化钒免疫层析试纸条,用于现场灵敏检测玉米中T-2毒素。通过超声辅助-液相剥离法合成碳化钒纳米片(vanadium carbide nanosheets,VCNSs),与T-2单克隆抗体(T-2 monoclonal antibody,T-2-mAb)偶联标记,作为信号探针。基于竞争原理,样品中的T-2毒素与检测线(T线)上T-2抗原竞争结合黑色的VCNSs-mAb探针,试纸条T线读值与T-2毒素质量浓度呈反比。以空白试纸条T线显色情况(信号读值>800,MD-600金标读数仪检测)和0.5、1.5 ng/mL T-2毒素的抑制率为优化指标,获得最佳实验条件,建立标准曲线。结果表明:该方法检测T-2毒素的线性范围在0.14~0.78 ng/mL之间、视觉检出限为0.25 ng/mL、消线值为1.5 ng/mL、半抑制浓度为0.344 ng/mL,灵敏度较传统胶体金试纸条提高了5.6 倍。此外,该方法与其他5 种常见真菌毒素均无交叉反应。针对玉米样品进行真实检测,回收率在92.8%~117.2%之间、变异系数均小于9.13%、检出限为5.06 μg/kg。综上所述,该方法具有操作简便、快速灵敏和成本低等优点,为谷物中T-2的检测提供一定的现场快速检测技术。

关键词: T-2毒素;玉米;碳化钒;免疫层析试纸条

Abstract: The purpose of this study was to construct a vanadium carbide-based immunochromatographic assay for the sensitive detection of T-2 toxin in maize. Vanadium carbide nanosheets (VCNSs) were synthesized by ultrasonic-assisted liquid phase exfoliation, and conjugated with T-2 monoclonal antibody (T-2-mAb) as a signal probe. Based on the competition principle, the T-2 toxin in samples competed with the T-2 antigen on the test (T) line for binding to the black VCNSs-mAb probe. As a result, the T-line value was inversely proportional to T-2 toxin concentration. The best experimental conditions and the standard curve were established based on color development in the T-line of the blank test strip (signal value > 800, detected using an MD-600 reader) and the inhibition rate of 0.5 and 1.5 ng/mL T-2 toxin. The results showed that the linear range of the proposed method was between 0.14 and 0.78 ng/mL, the visual detection limit was 0.25 ng/mL, the cut-off value was 1.5 ng/mL, and the half maximal inhibitory concentration (IC50) was 0.344 ng/mL. The sensitivity was 6.6 times higher than that of the traditional colloidal gold test strip. In addition, the assay did not cross-react with five other common mycotoxins. Moreover, the recoveries for real maize samples ranged from 92.8% to 117.2% with coefficients of variation (CV) of less than 9.13%, and the detection limit of 5.06 μg/kg. In summary, this method is characterized by simple operation, rapid sensitivity and low cost, which enables on-site rapid detection of T-2 toxin in cereals.

Key words: T-2 toxin; maize; vanadium carbide; immunochromatographic test strip

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