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首页> 外文期刊>Harmful Algae >Identification and quantification of the toxic dinoflagellate Gymnodinium sp. with competitive enzyme-linked immunosorbent assay (cELISA)
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Identification and quantification of the toxic dinoflagellate Gymnodinium sp. with competitive enzyme-linked immunosorbent assay (cELISA)

机译：鉴定和定量有毒的鞭毛鞭毛藻。竞争性酶联免疫吸附测定（cELISA）

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摘要

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 ln[A/(A_0 - A)]= 4.9193 -1.1006 log[cell amount] (R~2 = 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.
机译：产生了针对Gymnodinium sp。的多克隆抗体。通过用鼠源性菌株的细胞免疫兔子。基于物种特异性抗血清，开发了一种间接竞争性酶联免疫吸附测定（cELISA）来鉴定和定量Gymnodinium sp。建立了标准曲线，以在24至6,250,000个细胞之间的线性范围内，以对数为基础将cELISA信号与细胞数量相关联，推导的等式为ln [A /（A_0-A）] = 4.9193 -1.1006 log [细胞数量]（R〜2 = 0.9948，n = 5）。发现检测限为12个细胞。批内和批间变异系数（CV）分别为5.8％和9.7％。从中国胶州湾采集的野外样品用于评估该方法的鲁棒性。结果与光学显微镜下的细胞计数结果高度吻合。 cELISA的良好再现性和精密度表明该新技术可用于对Gymnodinium sp。进行快速定量。

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