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酶联免疫吸附法检测人血浆中的 Brevetoxin。

Detection of Brevetoxin in Human Plasma by ELISA.

机构信息

Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.

Harbor Branch Oceanographic Institute, Florida Atlantic University, Ft. Pierce, FL 34946, USA.

出版信息

J Anal Toxicol. 2022 Mar 21;46(3):322-327. doi: 10.1093/jat/bkab010.

Abstract

Florida red tides have become more common and persistent in and around the Gulf of Mexico. When in bloom, red tides can produce brevetoxins in high concentrations, leading to human exposures primarily through contaminated food and ocean spray. The research described here includes adapting and validating a commercial brevetoxin water test kit for human plasma testing. Pooled plasma was fortified with a model brevetoxin, brevetoxin 3, at concentrations from 0.00500 to 3.00 ng/mL to generate calibration curves and quality control samples. The quantitative detection range was determined to be 0.0400-2.00 ng/mL brevetoxin 3 equivalents with inter- and intraday accuracies ranging from 94.0% to 109% and relative standard deviations <20%, which is within the US Food and Drug Administration guidelines for receptor-binding assays. Additionally, cross-reactivity was tested using 4 of the 10 known brevetoxins and 12 paralytic shellfish toxins. The cross-reactivity varied from 0.173% to 144% for the commercially available brevetoxin standards and 0% for the commercially available paralytic shellfish toxin standards. Fifty individual unexposed human plasma samples were measured to determine the limit of detection and endogenous interferences to the test. The validated method was used to test 31 plasma samples collected from humans potentially exposed to brevetoxins, detecting 11 positives. This method has been proven useful to measure human exposure to brevetoxins and can be applied to future exposure events.

摘要

佛罗里达的赤潮在墨西哥湾及周边地区变得更加普遍和持久。当赤潮爆发时,它们会产生高浓度的麻痹性贝类毒素,导致人类主要通过受污染的食物和海洋喷雾接触到这些毒素。本研究包括对一种商业性麻痹性贝类毒素水质测试试剂盒进行适配和验证,以用于人类血浆检测。将混合血浆与模型麻痹性贝类毒素——短裸甲藻毒素 3 进行浓缩,浓度从 0.00500 到 3.00ng/mL,以生成校准曲线和质控样本。结果显示,定量检测范围为 0.0400-2.00ng/mL 的短裸甲藻毒素 3 当量,日内和日间精密度的准确率范围在 94.0%至 109%之间,相对标准偏差小于 20%,符合美国食品和药物管理局对受体结合分析的指导方针。此外,还使用 10 种已知的短裸甲藻毒素中的 4 种和 12 种麻痹性贝类毒素进行了交叉反应性测试。结果显示,市售短裸甲藻毒素标准品的交叉反应性在 0.173%至 144%之间,而市售麻痹性贝类毒素标准品的交叉反应性为 0%。检测了 50 份未暴露的人类血浆样本,以确定检测的检出限和内源性干扰。该验证方法用于检测 31 份可能接触过麻痹性贝类毒素的人类血浆样本,检测出 11 份阳性样本。该方法已被证明可用于测量人类接触麻痹性贝类毒素的情况,并可应用于未来的暴露事件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/affd/8679180/31a939bb3449/nihms-1761536-f0001.jpg

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