金纳米花免疫层析试纸条检测玉米中的呕吐毒素  

Detection of deoxynivalenol in corn using gold nanoflowers based immunochromatographic test strips

作　　者：王志华 赵一鸣 任文洁 王玉龙[2] 李一帆 曾峻涛 何保山[1] WANG Zhihua;ZHAO Yiming;REN Wenjie;WANG Yulong;LI Yifan;ZENG Juntao;HE Baoshan(National Engineering Laboratory,Provincal Key Laboratory of food Science Discipline,Henan University of Technology,Zhengzhou 450001,China;Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]河南工业大学粮油食品学院,河南工业大学国家工程实验室,省重点实验室,河南郑州450001 [2]江苏省农业科学院农产品质量安全与营养研究所,江苏南京210014

出　　处：《食品与发酵工业》2025年第2期293-299,共7页Food and Fermentation Industries

基　　金：国家重点研发计划项目(2023YFF1104600);河南省科技研发联合基金(225200810066);新疆农垦科学院重大科技项目(NCG202408)。

摘　　要：呕吐毒素(deoxynivalenol,DON)对人类和动物的健康以及食品经济构成了极大的威胁。该研究建立一种基于金纳米花(gold nanoflowers,AuNFs)的免疫层析试纸条(immunochromatographic test strip,ICTS)快速检测玉米中的呕吐毒素的方法。采用种子生长法制备了AuNFs,将其与抗DON单克隆抗体结合制备免疫层析试纸条中的金标探针,并对pH值、抗体量、上样缓冲液浓度、检测线(test-line,T线)浓度和金标探针量进行优化,最终用于玉米中的DON检测。经过优化实验条件,确定了最优pH值为7.5,最佳抗体添加量为1μL,最适上样缓冲液浓度为PBST+1%PVP K30,最好的T线质量浓度为0.1 mg/mL以及最佳探针量为3μL。在最优反应体系下,实验结果可以通过肉眼观察进行定性分析,该方法检出限为20μg/kg,灵敏度≥99%、假阳性率≤1%、假阴性率≤1%,且与现有标准方法检测结果一致,同时与其他真菌毒素无交叉反应。以AuNFs标记抗体为探针制备的ICTS灵敏度高,特异性强,可以实现玉米中DON的快速检测。Deoxynivalenol(DON)poses a great threat to human and animal health,as well as the food economy.This study established a method for the rapid detection of DON in corn using a gold nanoflowers(AuNFs)-based immunochromatographic test strips(ICTS).AuNFs were prepared using a seed growth method and conjugated with anti-DON monoclonal antibodies to generate the gold-labeled probes for immunochromatographic test strips.The pH,antibody volume,sample buffer concentration,T-line concentration,and the volume of gold-labeled probe were optimized.The developed method was subsequently used for detecting DON in corn.After optimizing the experimental conditions,the optimal pH was determined to be 7.5,the best antibody volume was 1μL,the most suitable sample buffer concentration was PBST+1%PVP K30,the optimal T-line concentration was 0.1 mg/mL,and the volume of gold-labeled probe was 3μL.Under the optimal conditions of the reaction system,the method achieved a detection limit of 20μg/kg,with a sensitivity of≥99%,a false positive rate of≤1%,and a false negative rate of≤1%.Additionally,the results were consistent with existing standard methods and show no cross-reactivity with other mycotoxins.The ICTS using the AuNFs-based immunochromatographic test strips exhibits high sensitivity and strong specificity,enabling rapid detection of DON in corn.

关 键 词：金纳米花 种子生长法 免疫层析试纸条 快速检测 呕吐毒素 

分 类 号：TS213.4[轻工技术与工程—粮食、油脂及植物蛋白工程]