JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2018 Vol.19 No.11 P.871-883

Dual flow immunochromatographic assay for rapid and simultaneous quantitative detection of ochratoxin A and zearalenone in corn, wheat, and feed samples

Author(s): Xian Zhang, Ke He, Yun Fang, Tong Cao, Narayan Paudyal, Xiao-feng Zhang, Hou-hui Song, Xiao-liang Li, Wei-huan Fang
Affiliation(s): China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, Zhejiang A&F University, Lin'an 311300, China; more
Corresponding email(s): whfang@zju.edu.cn
Key Words: Immunochromatographic assay, Gold nanoparticles, Ochratoxin A, Zearalenone, Quantification

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Abstract: A one-step dual flow immunochromatographic assay (DICGA), based on a competitive format, was developed for simultaneous quantification of ochratoxin A (OTA) and zearalenone (ZEN) in corn, wheat, and feed samples. The limit of detection for OTA was 0.32 ng/ml with a detection range of 0.53‒12.16 ng/ml, while for ZEN it was 0.58 ng/ml with a detection range of 1.06‒39.72 ng/ml. The recovery rates in corn, wheat, and feed samples ranged from 77.3% to 106.3% with the coefficient of variation lower than 15%. Naturally contaminated corn, wheat, and feed samples were analyzed using both DICGA and liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the correlation between the two methods was evaluated using a regression analysis. The DICGA method shows great potential for simple, rapid, sensitive, and cost-effective quantitative detection of OTA and ZEN in food safety control.

赭曲霉毒素A和玉米赤霉烯酮二联定量胶体金免疫层析试纸条的制备及应用

目的:优化建立二联胶体金免疫层析试纸条,实现对玉米、面粉和饲料中赭曲霉毒素A和玉米赤霉烯酮的同时快速定量检测.
创新点:采用纳米金颗粒标记真菌毒素单克隆抗体,基于竞争反应模式,通过对金颗粒尺寸、免疫层析相关组成材料及缓冲液配方的系列优化,同时借助手持式信号读取装备,实现对谷物和饲料中赭曲霉毒素A和玉米赤霉烯酮的同时定量检测.该方法简单、快速且成本低,与液相色谱串联质谱(LC-MS/MS)一致性良好,便于基层单位的推广使用.
方法:采用柠檬酸钠还原法制备纳米金颗粒并标记获得金标抗体.通过在基于竞争反应的免疫层析检测体系中,同时设置两条检测线(T1、T2)以实现两种毒素的同时检测,最终采用金标信号读数仪
实现定量检测.
结论:制备的二联定量胶体金试纸条对赭曲霉毒素A和玉米赤霉烯酮的检测限分别为0.32和0.58 ng/ml(图5),在玉米、小麦和饲料样本中的加标回收率为77.3%~106.3%,且变异系数均小于15%(表2).制备的二联定量胶体金免疫层析试纸条和LC-MS/MS对玉米、小麦和饲料等天然样本中赭曲霉毒素A和玉米赤霉烯酮的定量检测结果相关性较好(表3和图7),具有较好的应用价值.
关键词：免疫层析;纳米金颗粒;赭曲霉毒素A;玉米赤霉烯酮;定量检测

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[52]List of electronic supplementary materials

[53]Table S1 Optimal conditions of the dual flow immunochromatographic assay

[54]Table S2 Buffer types, optimum buffers, and optimum concentrations of additives in the dual flow immunochromatographic assay

[55]Table S3 Comparison of results of gold nanoparticles-based immunochromatographic assay published in the past

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赭曲霉毒素A和玉米赤霉烯酮二联定量胶体金免疫层析试纸条的制备及应用

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