Automated, high performance, flow-through chemiluminescence microarray for the multiplexed detection of phycotoxins
Automated, high performance, flow-through chemiluminescence microarray for the multiplexed detection of phycotoxins
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DOI：
10.1016/j.aca.2013.05.028
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发表时间：
2013-07-17
影响因子：
6.2
通讯作者：
Seidel, Michael
中科院分区：
文献类型：
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作者：
Szkola, Agathe;Campbell, Katrina;Seidel, Michael
建立了一种新的多路免疫分析法，用于贝类中藻毒素的分析。为此，建立了一种可再生化学发光芯片，可在MCR3分析平台上自动并行分析软骨藻酸（DA）、山田酸（OA）和蛤蚌毒素（STX）三种不同的藻毒素。作为一种测试格式，采用了间接竞争性免疫分析格式。这些藻毒素直接固定在环氧树脂活化的PEG芯片表面。平行分析是通过在一个微阵列芯片上同时添加所有分析物和特异性抗体来实现的。竞争反应结束后，用CCD相机记录CL信号。由于毒素芯片的再生能力，藻毒素的平行内部校准使用相同的芯片进行，这适用于25个连续测量。对3种目标藻毒素生成了多分析物校准曲线。在提取的贝类基质中，测定的DA、OA和STX的lod值分别为0.5 +/- 0.3 μ g -1、1.0 +/- 0.6 μ g -1和0.4 +/- 0.2 μ g -1，略低于PBS缓冲液。为了测定毒素回收率，对再生过程中观察到的信号损失进行了校正。经数学校正后，加标贝类样品在20 min内的DA、OA和STX的回收率分别为86.2%、102.5%和61.6%。这是基于抗体的藻毒素微阵列的首次演示。(C) 2013 Elsevier B.V.版权所有
A novel multiplexed immunoassay for the analysis of phycotoxins in shellfish samples has been developed. Therefore, a regenerable chemiluminescence (CL) microarray was established which is able to analyze automatically three different phycotoxins (domoic acid (DA), okadaic acid (OA) and saxitoxin (STX)) in parallel on the analysis platform MCR3. As a test format an indirect competitive immunoassay format was applied. These phycotoxins were directly immobilized on an epoxy-activated PEG chip surface. The parallel analysis was enabled by the simultaneous addition of all analytes and specific antibodies on one microarray chip. After the competitive reaction, the CL signal was recorded by a CCD camera. Due to the ability to regenerate the toxin microarray, internal calibrations of phycotoxins in parallel were performed using the same microarray chip, which was suitable for 25 consecutive measurements. For the three target phycotoxins multi-analyte calibration curves were generated. In extracted shellfish matrix, the determined LODs for DA, OA and STX with values of 0.5 +/- 0.3 mu g L-1, 1.0 +/- 0.6 mu g L-1, and 0.4 +/- 0.2 mu g L-1 were slightly lower than in PBS buffer. For determination of toxin recoveries, the observed signal loss in the regeneration was corrected. After applying mathematical corrections spiked shellfish samples were quantified with recoveries for DA, OA, and STX of 86.2%, 102.5%, and 61.6%, respectively, in 20 min. This is the first demonstration of an antibody based phycotoxin microarray. (C) 2013 Elsevier B.V. All rights reserved.