竞争性ELISA:一种准确、快速、有效的检测环境和生物样品中短链毒素的工具。

Jerome Naar, Allison Weidner, Daniel G Baden
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引用次数: 0

摘要

建立了一种用于短链毒素(PbTxs)分析的竞争性酶联免疫吸附试验(competitive ELISA)。短链毒素抗体与多步骤信号扩增程序相结合用于毒素检测。这个程序最大限度地减少非特定的信号和背景噪声,经常观察到复杂的矩阵。因此,可以对各种样品(海水,空气过滤器,哺乳动物体液,贝类等)进行分析,而不需要大量的提取和/或净化步骤。海水、尿液和血清等液体样品中的Brevetoxin分析无需预处理、稀释或纯化即可进行。在所测试的任何液体样品基质中,PbTxs的定量限为2 ng mL-1。对于贝类监测,分析是在贝类肉(5 g)与brevetoxin-ELISA缓冲液(200 mL)均质后进行的,可以在单个软体动物的组织上进行,也可以在贝类肉池上进行。在含有等量毒素(10 ng mL-1样品)的缓冲液、海水、哺乳动物体液和贝类匀浆中进行的PbTxs比较定量,差异不超过5%。这些数据表明,样品的基质组成不影响分析的性能。由于该检测不受基质组成的影响,可以在贝类匀浆中进行,因此该方法可用于预防或诊断人类暴露于PbTxs,并有可能取代目前用于监测贝类中PbTxs的小鼠生物检测。
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Competitive ELISA: An Accurate, Quick and Effective Tool to Monitor Brevetoxins in Environmental and Biological Sample.

A competitive Enzyme-Linked Immuno-Sorbent Assay (competitive ELISA) has been developed for analyzing brevetoxins (PbTxs). Antibodies to brevetoxins were used in combination with a multi-step signal amplification procedure for the detection of toxins. This procedure minimizes non-specific signals and background noise often observed in complex matrices. Therefore, analysis can be performed with various samples (seawater, air filter, mammalian body fluids, shellfish, etc.) without the need for extensive extraction and/or purification steps. Brevetoxin analysis in liquid samples like seawater, urine and serum can be performed without pretreatment, dilution or purification. The limit of quantification of PbTxs is 2 ng mL-1 in any of the liquid sample matrices tested. For shellfish monitoring, analyses are performed after homogenization of shellfish meat (5 g) with brevetoxin-ELISA buffer (200 mL) and can be performed on tissue from a single mollusk as well as on a pool of shellfish meat. Comparative quantification of PbTxs achieved in buffer, seawater, mammalian body fluid and shellfish homogenate spiked with equal amounts of toxin (10 ng mL-1 sample) varied by no more than 5%. These data suggest that the matrix composition of the sample does not affect the performance of the assay. Because this assay is not affected by matrix composition and can be performed in shellfish homogenate, this procedure can be used to prevent or diagnose human exposure to PbTxs and has the potential to replace the currently used mouse bioassay for monitoring PbTxs in shellfish.

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Brevetoxin Degradation and By-Product Formation via Natural Sunlight. Competitive ELISA: An Accurate, Quick and Effective Tool to Monitor Brevetoxins in Environmental and Biological Sample. Multi-Laboratory Study of Five Methods for the Determination of Brevetoxins in Shellfish Tissue Extracts. Florida Red Tide: Inhalation Toxicity of Karenia brevis Extract in Rats. An Epidemiologic Approach to the Study of Aerosolized Florida Red Tides.
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