食品科学 ›› 2022, Vol. 43 ›› Issue (18): 310-317.doi: 10.7506/spkx1002-6630-20210721-259

• 安全检测 • 上一篇    

高灵敏度磁荧光免疫层析试纸模式构建及在呕吐毒素检测中的应用

孙亚宁，杨苏珍，杨继飞，胡骁飞，陈鑫鑫，张颖硕，邓瑞广，张改平   

  1. （1.河南省农业科学院动物免疫学重点实验室，河南 郑州 450002；2.河南农业大学动物医学院，河南 郑州 450002）
  • 发布日期:2022-09-28
  • 基金资助:
    “十三五”国家重点研发计划重点专项（2018YFC1602903）；河南省农业科学院科技创新创意项目（2020CX20）； 中国博士后科学基金项目（2018M632778）；河南省农业科学院科研发展专项（2019CY04）

Development and Application of High-Sensitivity Magnetic Fluorescence Immunochromatographic Test Strip for Detection of Deoxynivalenol

SUN Yaning, YANG Suzhen, YANG Jifei, HU Xiaofei, CHEN Xinxin, ZHANG Yingshuo, DENG Ruiguang, ZHANG Gaiping   

  1. (1. Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; 2. College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China)
  • Published:2022-09-28

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摘要： 构建基于磁荧光纳米材料的免疫层析试纸模式，弥补现在免疫层析技术的不足，为更灵敏的免疫学快速检测提供技术支撑。以呕吐毒素（deoxynivalenol，DON）为靶标，采用溶剂热法制备羧基修饰的超顺磁颗粒，碳二亚胺法将磁颗粒、绿色荧光蛋白及DON单克隆抗体进行偶联，一步法制备磁荧光抗体探针，以DON人工抗原（DON-BSA）为检测线建立磁荧光免疫层析试纸。同时用胶体金标记DON单克隆抗体，以DON-BSA为检测线建立胶体金免疫层析试纸；制备的磁荧光抗体探针具有很好的磁性、荧光特性及抗体反应性，基于该探针成功制备了DON磁荧光免疫层析试纸，该试纸回归方程为y=－0.562x＋0.921，R2=0.990，IC50为5.611 ng/mL，检出限为1.089 ng/mL；制备了DON胶体金免疫层析试纸，该试纸裸眼检测灵敏度为500 ng/mL；定量检测回归方程为y=－0.543x＋1.485，R2=0.991，IC50为65.16 ng/mL，检出限为11.94 ng/mL。DON磁荧光免疫层析试纸的灵敏度是胶体金免疫层析试纸的10.96 倍。本实验建立的磁荧光免疫层析试纸模式可以同时实现样品的富集及荧光信号检测，提高检测灵敏度，并成功用于DON的检测，为磁荧光纳米颗粒广泛应用于免疫层析领域提供参考。

关键词: 磁荧光纳米颗粒；免疫学检测；呕吐毒素；试纸

Abstract: This study aimed to develop an immunochromatographic test strip model based on magnetic fluorescence nanomaterials in order to make up for the shortcoming of lateral flow immunoassay and provide technical support for the development of more sensitive rapid immunological assays. Using deoxynivalenol (DON) as the target, carboxyl-modified superparamagnetic particles were prepared by the solvothermal method. The magnetic particles, green fluorescent protein and anti-DON monoclonal antibody were coupled by the carbodiimide method to prepare magnetic fluorescent antibody probe. A magnetic fluorescence immunochromatographic test strip was established with DON-bovine serum albumin (BSA) as the test line. Meanwhile, a colloidal gold immunochromatographic test strip was established with colloidal gold-labeled anti-DON monoclonal antibody as the probe and DON-BSA as the test line. The magnetic fluorescence antibody probe exhibited good magnetic and fluorescent properties and antibody reactivity. The regression equation of the magnetic fluorescence immunochromatographic test strip was y = ?0.562x + 0.921, R2 = 0.990. The half maximal inhibitory concentration (IC50) was 5.611 ng/mL, and the detection limit was 1.089 ng/mL. The sensitivity of the colloidal gold immunochromatographic test strip was 500 ng/mL with naked eyes, the regression equation was y = ?0.543x + 1.485, R2 = 0.991, the IC50 was 65.16 ng/mL, and the detection limit was 11.94 ng/mL. The sensitivity of the magnetic fluorescence immunochromatographic test strip was 10.96 folds higher than that of the colloidal gold immunochromatographic test strip. The magnetic fluorescence immunochromatographic test strip allowed simultaneous sample enrichment and fluorescence signal detection and was successfully used in the detection of DON with high sensitivity. This study may lay the foundation for the application of magnetic fluorescence nanoparticles in the field of immunochromatographic assay.

Key words: magnetic fluorescent nanoparticles; immunoassay; deoxynivalenol; test strip

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