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Enzyme-linked immunosorbent assay and colloidal gold immunoassay for ochratoxin A: investigation of analytical conditions and sample matrix on assay performance

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Abstract

A polyclonal antibody against ochratoxin A (OTA) was produced from rabbits immunized with the OTA–BSA conjugate. A competitive direct enzyme-linked immunosorbent assay (cdELISA) and a membrane-base colloidal gold immunoassay in flow-through format were developed for the rapid detection of OTA in various food matrices. In the cdELISA, the concentration causing 50% inhibition was 0.07 ng mL−1, and the effects of different chemical conditions (ionic strength, pH value, and organic solvent) were studied. The sensitivity of the assay was higher than those previously reported. A simple, rapid, and efficient extraction method was developed and 74–110% recoveries of spiked samples were obtained. Fifty percent methanol extracts of some food samples such as barley, wheat, oat, corn, rice, and raisins could be analyzed directly by immunoassay after dilution in PBS; grape juice and beer samples could be analyzed directly after dilution with PBS; for coffee samples, a more complex method was used to remove the matrix effect effectively. Membrane-based colloidal gold immunoassays had a visual detection limit of 1.0 ng mL−1 for OTA with a detection time of less than 10 min. For the validation of the cdELISA and membrane-based colloidal gold immunoassay, samples were analyzed by high-performance liquid chromatography. The correlation between data obtained using the microwell assay and HPLC was good (R 2 = 0.984). The developed immunoassay methods are suitable for the rapid quantitative or qualitative determination of OTA in food samples.

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Acknowledgements

The authors are grateful for financial supports from the Ministry of Science and Technology of the People’s Republic of China (project No.2006BAD05A06, 2006AA10Z448), the New Century Talent Program of Ministry of Education of People’s Republic of China (project No. NECT-04-0243).

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Authors and Affiliations

  1. Tianjin Key Laboratory of Food Nutrition and Safety, Faculty of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China

    Xiang-Hong Wang, Tao Liu, Na Xu, Yan Zhang & Shuo Wang

  2. Faculty of Food Science and Technology, Agricultural University of Hebei, Baoding, 071001, China

    Xiang-Hong Wang

Authors
  1. Xiang-Hong Wang
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  2. Tao Liu
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  3. Na Xu
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  4. Yan Zhang
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  5. Shuo Wang
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Correspondence to Shuo Wang.

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Wang, XH., Liu, T., Xu, N. et al. Enzyme-linked immunosorbent assay and colloidal gold immunoassay for ochratoxin A: investigation of analytical conditions and sample matrix on assay performance. Anal Bioanal Chem 389, 903–911 (2007). https://doi.org/10.1007/s00216-007-1506-6

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  • DOI: https://doi.org/10.1007/s00216-007-1506-6

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