Identification and quantification of the toxic dinoflagellate Gymnodinium sp. with competitive enzyme-linked immunosorbent assay (cELISA) |
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| Authors: | ZeYu Xin ZhiGang Yu TanChun Wang Xin Hui WanLi Gou Jun Sun Haigang Qi RongXiu Li |
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| Institution: | a College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266003, PR China;b School of Life Science and Biotechnology, Shanghai Jiao tong University, Shanghai 200030, PR China;c Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Graduate School of the Chinese Academy of Sciences, Shanghai 200031, PR China;d College of Marine Life Sciences, Ocean University of China, Qingdao 266003, PR China |
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| Abstract: | Polyclonal antibodies were raised against Gymnodinium sp. by immunizing rabbits with cells of the axenic strain. Based on the species-specific antiserum, an indirect competitive enzyme-linked immunosorbent assay (cELISA) was developed to identify and quantify Gymnodinium sp. A standard curve was established to correlate the cELISA signal to cell amount on a logit-log basis in the linear range between 24 and 6,250,000 cells, and the equation deducted was lnA/(A0 − A)]= 4.9193 − 1.1006 logcell amount] (R2 = 0.9948, n = 5). The detection limit was found to be 12 cells. The intra-assay and inter-assay coefficients of variation (CVs) were 5.8% and 9.7%, respectively. Field samples collected from Jiaozhou Bay, China were used to assess the robustness of the method. The results showed high agreement with that of cell-counting with a light microscope. The good reproducibility and precision of the cELISA implied that this new technique could be used for fast quantification of Gymnodinium sp. |
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| Keywords: | Antibody Competitive ELISA Gymnodinium sp Quantification |
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