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Development of an immunochromatographic strip test for the rapid detection of okadaic acid in shellfish sample

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Abstract

A rapid detection technology for okadaic acid (OA) in shellfish with one-step immunochromatographic assay using colloidal gold-labeled monoclonal antibody (Mab) probe was developed. OA is one of the diarrhetic shellfish toxins. Firstly, OA was conjugated to bovine serum albumin, and the conjugations as immunogen were injected into mice to raise the polyclonal antibody against OA. Hybridoma cells fused between spleen cells from immunized mouse and myeloma cells (Sp2/0) were prepared and injected into mice intraperitoneally at 1 × 106 cells to produce monoclonal antibody in the ascitic fluid. With the monoclonal antibody against OA, the idc-ELISA assay was established to detect OA. The calibration curve for OA was linear over the concentration range of 0.31–50 ng mL−1, and the detection limit for OA was 0.45 ng mL−1. On that basis, paper test strips for detecting OA were prepared, and a fast detection method for okadaic acid using gold-labeled immunological assay was established. With the paper test strips, the detection limit was 6.25 ng mL−1, and whole detection process for OA in shellfish samples needed only about 40 min.

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Acknowledgments

This study was supported by the China National Marine 863 Program (2007AA09Z406), the Project for Excellent Academic Leaders (08SD14037), the International Cooperation Project (08540702600), the Biomedical Sciences Project (10391901900), and the Mountaineering Project (06dz12015) from Shanghai Committee for Science and Technology, the Hydrobiological Project of Shanghai Leading Academic Discipline (III) (S30701), China.

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Authors and Affiliations

  1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China

    Leqin Hu, Junxiu Liu, Qing Wang, Yang Zhang, Rui Jia, Chuner Cai, Weining Wu & Peimin He

  2. Institutes for Marine Science Research, Shanghai Ocean University, Shanghai, 201306, China

    Leqin Hu, Junxiu Liu, Qing Wang, Yang Zhang, Rui Jia, Chuner Cai, Weining Wu & Peimin He

  3. School of Biological Sciences, The University of Hong Kong, Hong Kong, Hong Kong, China

    Steven-Feng Chen

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  1. Leqin Hu
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Correspondence to Peimin He.

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Hu, L., Liu, J., Wang, Q. et al. Development of an immunochromatographic strip test for the rapid detection of okadaic acid in shellfish sample. J Appl Phycol 25, 1091–1099 (2013). https://doi.org/10.1007/s10811-012-9949-3

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  • DOI: https://doi.org/10.1007/s10811-012-9949-3

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