Development of an immunochromatographic strip test for the rapid detection of okadaic acid in shellfish sample
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Development of an immunochromatographic strip test for the rapid detection of okadaic acid in shellfish sample Leqin Hu & Junxiu Liu & Qing Wang & Yang Zhang & Rui Jia & Chuner Cai & Weining Wu & Steven-Feng Chen & Peimin He
Received: 15 April 2012 / Revised and accepted: 15 November 2012 / Published online: 24 February 2013 # Springer Science+Business Media Dordrecht 2012
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. L. Hu : J. Liu : Q. Wang : Y. Zhang : R. Jia : C. Cai : W. Wu : P. He (*) College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China e-mail: [email protected] L. Hu : J. Liu : Q. Wang : Y. Zhang : R. Jia : C. Cai : W. Wu : P. He Institutes for Marine Science Research, Shanghai Ocean University, Shanghai 201306, China S.-F. Chen School of Biological Sciences, The University of Hong Kong, Hong Kong, Hong Kong, China
Keywords Diarrhetic shellfish poison (DSP) . Okadaic acid (OA) . Antibody . Enzyme-linked immunosorbent assay (ELISA) . Colloidal gold, paper test strip, fast detection
Introduction Okadaic acid (OA) is a lipophilic marine algal toxin, which was first extracted from the sponges Halichondria okadai and Halichondria melanodocia (Tachibana et al. 1981). Later it was found to be the same toxin as that isolated from the dinoflagellates Prorocentrum and Dinophysis (Murakami et al. 1982; Carmody et al. 1996). OA is one of the major toxins responsible for diarrhetic shellfish poisoning (DSP) (Murata et al. 1982) and can be accumulated in shellfish feeding on toxigenic algae cells. Diarrhetic shellfish poisoning is the term used to describe the rapid onset of gastrointestinal symptoms such as vomiting and diarrhea in people who consume toxic shellfish (Amzil et al. 1992; Souto et al. 2001). In addition, OA potentially can facilitate tumor promotion. However, the medical treatment for aforementioned symptoms has not been confirmed. Treatment with strong mineral acids could destroy OA in the
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