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一种用于定量检测冈田酸的独特型-抗独特型竞争性免疫分析方法。

An idiotypic-anti-idiotypic competitive immunoassay for quantitation of okadaic acid.

作者信息

Shestowsky W S, Quilliam M A, Sikorska H M

机构信息

Rougier Bio-Tech Ltd, Montreal, Quebec, Canada.

出版信息

Toxicon. 1992 Nov;30(11):1441-8. doi: 10.1016/0041-0101(92)90519-b.

Abstract

A competitive indirect enzyme-linked immunosorbent assay for the measurement of okadaic acid, a marine toxin, was developed. The assay uses a murine monoclonal anti-idiotypic antibody bearing an internal image of okadiac acid epitope to capture an anti-okadaic acid monoclonal antibody in the presence of free okadaic acid. Bound anti-okadaic acid antibody is detected with peroxidase-conjugated anti-mouse immunoglobulin antiserum. If present, free toxin will lessen the amount of anti-okadaic acid antibody binding to its corresponding anti-idiotypic antibody in a dose-dependent manner that can be quantified from the standard curve. The assay permits reliable measurement of okadaic acid in the 9-81 ng/ml range. The intra- and interassay coefficients of variation in the measurement of OA in the toxin spiked mussel samples averaged 9% and 12%, respectively. The assay is rapid, accurate, reproducible and relatively simple to perform. It may be of potential use to laboratories involved in monitoring the toxin levels in plankton, seafood or sponges.

摘要

开发了一种用于测定海洋毒素冈田酸的竞争性间接酶联免疫吸附测定法。该测定法使用带有冈田酸表位内影像的鼠单克隆抗独特型抗体,在游离冈田酸存在的情况下捕获抗冈田酸单克隆抗体。用辣根过氧化物酶标记的抗小鼠免疫球蛋白抗血清检测结合的抗冈田酸抗体。如果存在游离毒素,它将以剂量依赖的方式减少抗冈田酸抗体与其相应抗独特型抗体的结合量,这可以从标准曲线中定量。该测定法能够可靠地测定9 - 81 ng/ml范围内的冈田酸。在添加毒素的贻贝样品中测定OA时,批内和批间变异系数平均分别为9%和12%。该测定法快速、准确、可重复且操作相对简单。它可能对参与监测浮游生物、海鲜或海绵中毒素水平的实验室有潜在用途。

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