Ramón-Azcón Javier, Kunikata Ryouta, Sanchez F-J, Marco M-P, Shiku Hitoshi, Yasukawa Tomoyuki, Matsue Tomokazu
Applied Molecular Receptors Group, IIQAB-CSIC, Barcelona, Spain.
Biosens Bioelectron. 2009 Feb 15;24(6):1592-7. doi: 10.1016/j.bios.2008.08.035. Epub 2008 Aug 28.
The detection of atrazine using a novel optical immunosensing technique based on negative dielectrophoresis (n-DEP) in microfluidic channels is described. Atrazine is a toxic triazine herbicide within the most frequently used. Polystyrene microparticles (6 microm diameters) modified with bovine serum albumin conjugated with atrazine (atrazine-BSA) were manipulated and captured when subjected to intense n-DEP electric fields. Specifically, particles were trapped when AC voltages with amplitudes of 10 V(peak) and frequencies over 1 MHz were applied to the electrodes. The immunological reaction occurring on the particles for detecting atrazine is based on an indirect competitive assay using a secondary anti-mouse immunogloburin G (IgG) antibody labeled with fluorescein isothiocyanate. The microfluidic device, with three-dimensional microelectrodes, was fabricated comprising two caged areas, allowing two simultaneous measurements inside the same microfluidic channel. The performance of this n-DEP immunosensing technique was evaluated using wine samples. The immunodevice showed a limit of detection for atrazine in buffer samples of 0.11 microgL(-1) and in pre-treated wine samples of 6.8 microg L(-1); these detection limits are lower than the maximum residue level (MRL) established by the European Community for residues of this herbicide in wine (50 microg L(-1)). This methodology offers great promise for rapid, simple, cost effective, and on-site analysis of biological, foods and beverages, and environmental samples.
本文描述了一种基于负介电泳(n-DEP)的新型光学免疫传感技术在微流控通道中检测阿特拉津的方法。阿特拉津是一种常用的有毒三嗪类除草剂。用与阿特拉津偶联的牛血清白蛋白修饰的聚苯乙烯微粒(直径6微米)在强n-DEP电场作用下被操控和捕获。具体而言,当向电极施加幅度为10 V(峰值)且频率超过1 MHz的交流电压时,微粒会被捕获。基于间接竞争分析,利用异硫氰酸荧光素标记的抗小鼠免疫球蛋白G(IgG)二抗在微粒上发生检测阿特拉津的免疫反应。该微流控装置带有三维微电极,制作而成的装置包含两个笼形区域,可在同一微流控通道内同时进行两次测量。使用葡萄酒样品评估了这种n-DEP免疫传感技术的性能。该免疫检测装置对缓冲液样品中阿特拉津的检测限为0.11微克/升,对预处理葡萄酒样品中阿特拉津的检测限为6.8微克/升;这些检测限低于欧洲共同体规定的该除草剂在葡萄酒中的最大残留限量(MRL,50微克/升)。这种方法对于生物、食品和饮料以及环境样品的快速、简单、经济高效的现场分析具有很大的前景。