Quantitative microarray pesticide analysis
Quantitative microarray pesticide analysis
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DOI：
10.1016/j.jim.2004.01.004
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发表时间：
2004-03-01
影响因子：
2.2
通讯作者：
Christensen, CBV
中科院分区：
文献类型：
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作者：
Belleville, E;Dufva, M;Christensen, CBV
为了取代基于微量滴定板的农药免疫测定法，我们已经开发了一种定量的竞争性微阵列免疫测定法，其允许快速和高度灵敏的定量的草腈降解产物2，6-二氯苯甲酰胺（BAM），和突出使用的除草剂阿特拉津。农药分析是基于荧光偶联单克隆抗体（mAb）与其各自分析物的竞争性结合。最低检测限计算为1纳克/升（5 pM）的BAM和3纳克/升（10 pM）的阿特拉津。相应的IC 50值分别为BAM 10 ng/l（50 pM）和莠去津34 ng/l（160 pM）。与现有的微量滴定板免疫测定相比，发现微阵列的灵敏度高出20倍。与在超过1000倍浓缩样品上进行的气相色谱-质谱（GCMS）分析相比，基于微阵列的免疫测定使用非浓缩样品的灵敏度甚至高出10倍。与单一分析物定量相比，同时测量两种分析物不影响测定灵敏度。除了灵敏度的增加和多重定量的可能性之外，与微量滴定板免疫测定和GCMS相比，微阵列平台的测定时间和测定复杂性显著降低，表明基于微阵列的免疫测定是用于测量皮摩尔量的分析物（例如临床相关分析物）的可行方法。(C)2004 Elsevier B. V.保留所有权利。
To replace a pesticide immunoassay based on microtiter plates, we have developed a quantitative, competitive microarray immunoassay, which permits rapid and highly sensitive quantification of the dichlobenil degradation product 2,6-dichlorobenzamide (BAM), and the prominently used herbicide atrazine. The pesticide analysis is based on the competitive binding of fluorescence conjugated monoclonal antibodies (mAb) to their respective analytes. Lowest detection limits were calculated to 1 ng/l (5 pM) for BAM and 3 ng/l (10 pM) for atrazine. Corresponding IC50 values were, 10 ng/l (50 pM) for BAM and 34 ng/l (160 pM) for atrazin, respectively. In comparison to the existing microtiter plate immunoassay, the microarray was found to be up to 20-fold more sensitive. Compared to the gas chromatography with mass spectroscopy (GCMS) analysis performed on more than 1000-fold concentrated samples, the microarray-based immunoassay was even 10-fold more sensitive using non-concentrated samples. Measuring both analytes simultaneously did not affect assay sensitivity compared to single analyte quantification. Besides a gain in sensitivity and the possibility of multiplex quantification, assay times and assay complexity were reduced drastically with the microarray platform compared to the rnicrotiter plate immunoassay and GCMS, suggesting that the microarray based immunoassay is a viable method for measuring picomolar amounts of analytes, e.g. clinically relevant analytes. (C) 2004 Elsevier B.V. All rights reserved.