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条带式免疫分析快速检测噻苯达唑。

Strip-based immunoassay for rapid detection of thiabendazole.

机构信息

Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, Institute of Chemical Technology, Technická 3, 16628 Prague, Czech Republic.

出版信息

Biosens Bioelectron. 2010 May 15;25(9):2122-8. doi: 10.1016/j.bios.2010.02.011. Epub 2010 Feb 24.

Abstract

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 IC(50) 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.

摘要

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。本研究报告了一种基于间接竞争原理的噻菌灵检测免疫层析检测方法的开发,该方法使用碳颗粒作为标记物。硝酸纤维素膜条在规定的测试区域涂有噻菌灵-蛋白质结合物。在抗体-碳复合物和噻菌灵沿条带流动的过程中,测试线上形成的黑色强度反映了噻菌灵的浓度,可以进行半定量目测估计。优化后的测试在 10 分钟内完成,标准样品的目视检测限达到 0.25ng/mL。此外,还使用扫描密度计对免疫层析进行了定量评估。基于标准曲线,该检测方法的检测限低至 0.08+/-0.03ng/mL,IC(50)值为 0.60+/-0.08ng/mL,线性工作范围为 0.11-4.13ng/mL。测试精度、稳定性和特异性的结果表明,该测定法提供了可靠的性能。该免疫层析试纸条用于分析浓度在 0.05-5mg/L 范围内的加标果汁。通过简单稀释样品可以避免基质干扰。目视和仪器评估都表明与 ELISA 结果具有良好的一致性。果汁的回收率在 81.9%至 123.6%之间,相对标准偏差范围在 9.9%至 19.3%之间。开发的条带提供了一种有用的快速简便的方法,可用于在远远低于最大残留限量的水平下筛选果汁中的噻菌灵。

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