Journal of Zhejiang University-SCIENCE B

, Volume 19, Issue 11, pp 871–883 | Cite as

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

  • Xian Zhang
  • Ke He
  • Yun Fang
  • Tong Cao
  • Narayan Paudyal
  • Xiao-feng Zhang
  • Hou-hui Song
  • Xiao-liang Li
  • Wei-huan FangEmail author


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.

Key words

Immunochromatographic assay Gold nanoparticles Ochratoxin A Zearalenone Quantification 




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。


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。


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。


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。


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


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Copyright information

© Zhejiang University and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.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 TechnologyZhejiang A&F UniversityLin’anChina
  2. 2.Zhejiang University Institute of Preventive Veterinary MedicineZhejiang Provincial Key Laboratory of Preventive Veterinary MedicineHangzhouChina
  3. 3.Zhejiang Academy of Science and Technology for Inspection and QuarantineHangzhouChina

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