Sandwich ELISA Assay for the Quantitation of Palytoxin and Its Analogs in Natural Samples
Sandwich ELISA Assay for the Quantitation of Palytoxin and Its Analogs in Natural Samples
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DOI：
10.1021/es304222t
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发表时间：
2013-02-19
影响因子：
11.4
通讯作者：
Tubaro, A.
中科院分区：
文献类型：
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作者：
Boscolo, S.;Pelin, M.;Tubaro, A.
海葵毒素是一种强效的海洋生物毒素，最近已成为地中海特有的，并且越来越频繁地与海鲜有关。由于它们的高毒性，需要合适的方法来定量海葵毒素。因此，我们建立了一个间接的夹心ELISA的海葵毒素和42-羟基-海葵毒素。一项实验室内研究显示，（检测限，LOD = 1.1 ng/mL;定量限，LOQ = 2.2 ng/mL），准确度（偏倚为2.1%），重复性（试验内和试验间变异性的RSDr分别= 6%和9%）和特异性：其它常见的海洋毒素（冈田酸、软骨藻酸、石房蛤毒素、短尾蛤毒素-3和耶索毒素）在该测定中不发生交叉反应。它在三种不同的基质中表现良好：贻贝提取物、藻网样品和海水的LOQ分别为11.0、9.6和2.4 ng/mL，具有良好的准确度和精密度。海产品中的定量限为11 μ g海葵毒素/kg贻贝肉，低于最常用的检测技术LC-MS/MS。
Palytoxins are potent marine biotoxins that have recently become endemic to the Mediterranean Sea, and are becoming more frequently associated with seafood. Due to their high toxicity, suitable methods to quantify palytoxins are needed. Thus, we developed an indirect sandwich ELISA for palytoxin and 42-hydroxy-palytoxin. An intra-laboratory study demonstrated sensitivity (limit of detection, LOD = 1.1 ng/mL; limit of quantitation, LOQ = 2.2 ng/mL), accuracy (bias of 2.1%), repeatability (RSDr = 6% and 9% for intra- and interassay variability, respectively) and specificity: other common marine toxins (okadaic acid, domoic acid, saxitoxin, brevetoxin-3, and yessotoxin) do not cross-react in this assay. It performed well in three different matrices: observed LOQs were 11.0, 9.6, and 2.4 ng/mL for mussel extracts, algal net samples and seawater, respectively, with good accuracy and precision. The LOQ in seafood is 11 mu g palytoxin/kg mussel meat, lower than that of the most common detection technique, LC-MS/MS.