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用于检测软骨藻酸、冈田酸和石房蛤毒素的 ELISA 方法的开发及其在检测比利时采集的贝类毒素样品中的适用性。

Development of ELISAs for detecting domoic acid, okadaic acid, and saxitoxin and their applicability for the detection of marine toxins in samples collected in Belgium.

机构信息

Departement Sante, CERgroupe, B-6900 Marloie, Belgium.

出版信息

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2010 Jun;27(6):859-68. doi: 10.1080/19440041003662881.

Abstract

Okadaic acid, a diarrhetic shellfish poison, domoic acid, an amnesic shellfish poison, and saxitoxin, a paralytic shellfish poison, are three of the best-known marine biotoxins. The mouse bioassay is the method most widely used to detect many of these toxins in shellfish samples, but animal welfare concerns have prompted researchers to seek alternative methods of detection. In this study, three direct competitive enzyme-linked immunosorbent assays (ELISAs), each based on antibodies raised in rabbits against a conjugate of the analyte of interest, were developed for marine biotoxin detection in mussel, oyster, and scallop. One assay was for okadaic acid, one for saxitoxin, and one for domoic acid usually detected and quantified by high-performance liquid chromatography-ultraviolet light (HPLC-UV). All three compounds and a number of related toxins were extracted quickly and simply from the shellfish matrices with a 9 : 1 mixture of ethanol and water before analysis. The detection capabilities (CCbeta values) of the developed ELISAs were 150 microg kg(-1) for okadaic acid, 50 microg kg(-1) for domoic acid, and 5 microg kg(-1) or less for saxitoxin. The assays proved satisfactory when used over a 4-month period for the analysis of 110 real samples collected in Belgium.

摘要

冈田酸、软骨藻酸(一种失忆性贝类毒素)和石房蛤毒素(一种麻痹性贝类毒素)是三种最为知名的海洋生物毒素。在贝类样本中检测这些毒素的方法中,鼠生物测定法是最广泛应用的方法,但动物福利问题促使研究人员寻求替代的检测方法。在本研究中,我们开发了三种基于兔抗分析物结合物的直接竞争酶联免疫吸附测定法(ELISA),用于贝类中海洋生物毒素的检测。其中一个检测法针对冈田酸,一个针对石房蛤毒素,一个针对软骨藻酸(通常通过高效液相色谱-紫外光法(HPLC-UV)检测和定量)。所有三种化合物和一些相关毒素在分析前用 9:1 的乙醇和水混合物从贝类基质中快速而简单地提取。开发的 ELISA 的检测能力(CCbeta 值)分别为:冈田酸 150μgkg(-1),软骨藻酸 50μgkg(-1),石房蛤毒素 5μgkg(-1)或更低。这些检测方法在比利时采集的 110 个实际样本进行为期 4 个月的分析期间表现令人满意。

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