Strip-based immunoassay for rapid detection of thiabendazole
Strip-based immunoassay for rapid detection of thiabendazole
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DOI：
10.1016/j.bios.2010.02.011
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发表时间：
2010-05-15
影响因子：
12.6
通讯作者：
Fukal, Ladislav
中科院分区：
文献类型：
--
作者：
Blazkova, Martina;Rauch, Pavel;Fukal, Ladislav
人们越来越关注研究和实施快速筛选方法来检测农药残留。本文报道了一种基于间接竞争原理，以碳颗粒为标记物的噻菌灵免疫试纸条检测方法。在规定的试验区用噻菌灵-蛋白质结合物包被硝酸纤维素膜条。在抗体-碳复合物和噻菌灵沿着条带流动时，在测试线中形成的黑色强度反映了噻菌灵浓度，并且可以目视进行半定量估计。优化后的方法在10 min内完成，目视检测限达到0.25 ng/mL标准样品。此外，使用扫描光密度法定量评价免疫条。根据标准曲线，拟定检测的检测限低至0.08 +/- 0.03 ng mL（-1），IC 50值为0.60 +/- 0.08 ng mL（-1），线性工作范围为0.11-4.13 ng mL（-1）。精密度、稳定性和特异性测试结果表明，该方法性能可靠，可用于0.05-5 mg L-1果汁样品的检测。通过简单稀释样品避免了基质干扰。目视和仪器评价均表明与ELISA获得的结果具有良好的一致性。果汁中的回收率为81.9 - 123.6%，相对标准偏差为9.9 - 19.3%。开发的试纸条提供了潜在的作为一个有用的快速和简单的方法筛选水平远低于最大残留限量的果汁中噻菌灵。(C)2010 Elsevier B. V.保留所有权利。
There is increased interest in the investigation and implementation of rapid screening methods for detection of pesticide residues. This study reports development of an immunostrip test for thiabendazole detection based on indirect competitive principle using carbon particles as a label. Nitrocellulose membrane strip was coated with a thiabendazole-protein conjugate in the defined test zone. In flow of an antibody-carbon complex and thiabendazole along the strip, the intensity of black colour formed in the test line reflected the thiabendazole concentration and semi-quantitative estimation could be carried out visually. The optimized test was accomplished within 10 min and the visual detection limit was achieved 0.25 ng mL(-1) of standard sample. Moreover, immunostrip was evaluated quantitatively using scanning densitometry. Based on standard curve, the detection limit of the proposed test was as low as 0.08 +/- 0.03 ng mL(-1) with an IC50 value of 0.60 +/- 0.08 ng mL(-1) and a linear working range of 0.11-4.13 ng mL(-1). Results of testing precision, stability, and specificity demonstrated that the assay provided a reliable performance.This immunostrip was applied to analysis of spiked fruit juices in range of 0.05-5 mg L-1. Matrix interferences were avoided by simple dilution of samples. Both visual and instrumental evaluations indicated a good agreement with results obtained by ELISA. Recoveries from juices were from 81.9 to 123.6% and relative standard deviations ranged from 9.9 to 19.3%. The developed strip offers potential as a useful rapid and simple method for screening of thiabendazole in fruit juices at levels far below the maximum residue limits. (C) 2010 Elsevier B.V. All rights reserved.