碳化钒免疫层析法检测玉米中T-2毒素  

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

作　　者：罗昌伟 陈晓阳 黄现青[1] 宋莲军[1] 张西亚 补彤 LUO Changwei;CHEN Xiaoyang;HUANG Xianqing;SONG Lianjun;ZHANG Xiya;BU Tong(School of Food Science and Technology,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学食品科学技术学院,河南郑州450002

出　　处：《食品科学》2024年第19期227-234,共8页Food Science

基　　金：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)。

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5.06μg/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。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 crossreact 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 5.06μg/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.

关 键 词：T-2毒素 玉米 碳化钒 免疫层析试纸条 

分 类 号：TS201.6[轻工技术与工程—食品科学]