Congener-independent immunoassay for microcystins and nodularins
Congener-independent immunoassay for microcystins and nodularins
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DOI：
10.1021/es011182f
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发表时间：
2001-12-15
影响因子：
11.4
通讯作者：
Dietrich, DR
中科院分区：
文献类型：
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作者：
Fischer, WJ;Garthwaite, I;Dietrich, DR
蓝细菌（蓝绿藻）（例如，微囊藻和节球藻属）在世界各地的淡水和半咸水中发现了能够产生有毒肽的细菌。这些毒素包括微囊藻毒素（MC）七肽（> 60个同源物）和节球藻毒素五肽（约100个同源物）。5同系物）。蓝藻环肽毒素对人类、其他哺乳动物、鸟类和鱼类有害。急性暴露于高浓度的这些毒素会导致肝损伤，而亚慢性或慢性暴露可能会促进肝肿瘤的形成。在地表水和饮用沃茨中检测环肽蓝藻毒素受到检测限低的限制，目前的常规检测仅限于少数同类物，即不常见的β-氨基酸ADDA（4 E，6 E-3-氨基-9-甲氧基-2，6，8-三甲基-1-10-苯基癸-4，6-二烯酸）存在于大多数（> 80%）已知的有毒五肽和七肽毒素同系物中。在这里，我们报告的两个ADDA半抗原的合成，ADDA抗体的提高，并利用这些抗体检测微囊藻毒素和节球藻毒素的竞争性间接ELISA的发展。该检测方法的定量限为0.02-0.07 ng/mL（取决于存在的同系物），低于世卫组织提出的饮用水指南（1 ng/mL），与样品基质（原水、饮用水或PBS中的纯毒素）无关。这种新的ELISA是强大的，可以在没有样品预浓缩，检测淡水样品中的毒素在较低的浓度比蛋白磷酸酶抑制试验，并显示出非常好的交叉反应性与所有蓝藻环肽毒素同源物测试到目前为止（MC-LR，-RR，-YR，-LW，-LF，3-desmethyl-MC-LR，3-desmethyl-MIC-RR，和nodularin）。
Cyanobacteria (blue-green algae) (e.g., Microcystis and Nodularia spp.) capable of producing toxic peptides are found in fresh and brackish water worldwide. These toxins include the microcystin (MC) heptapeptides (> 60 congeners) and the nodularin pentapeptides (ca. 5 congeners). Cyanobacterial cyclic peptide toxins are harmful to man, other mammals, birds, and fish. Acute exposure to high concentrations of these toxins causes liver damage, while subchronic or chronic exposure may promote liver tumor formation. The detection of cyclic peptide cyanobacterial toxins in surface and drinking waters has been hampered by the low limits of detection required and that the present routine detection is restricted to a few of the congeners, The unusual beta -amino acid ADDA (4E,6E-3-amino-9-mothoxy-2,6,8-trimethyl-1-10-phenyldeca-4,6-dienoic acid) is present in most (> 80%) of the known toxic penta- and heptapeptide toxin congeners. Here, we report the synthesis of two ADDA-haptens, the raising of antibodies to ADDA, and the development of a competitive indirect ELISA for the detection of microcystins and nodularins utilizing these antibodies. The assay has a limit of quantitation of 0.02-0.07 ng/mL (depending on which congeners are present), lower than the WHO-proposed guideline (1 ng/mL) for drinking water, irrespective of the sample matrix (raw water, drinking water, or pure toxin in PBS). This new ELISA is robust, can be performed without sample preconcentration, detects toxins in freshwater samples at lower concentrations than does the protein phosphatase inhibition assay, and shows very good cross-reactivity with all cyanobacterial cyclic peptide toxin congeners tested to date (MC-LR, -RR, -YR, -LW, -LF, 3-desmethyl-MC-LR, 3-desmethyl-MIC-RR, and nodularin).