INQUISAL, Department of Chemistry, National University of San Luis, CONICET, Chacabuco 917, D5700BWS, San Luis, Argentina.
Anal Biochem. 2011 Feb 1;409(1):98-104. doi: 10.1016/j.ab.2010.10.015. Epub 2010 Oct 14.
This article describes a microfluidic immunosensor, developed for the detection of IgG antibodies specific to Echinococcus granulosus in human serum samples, which represents an alternative tool that can be used for the immunodiagnosis of hydatidosis in an automated way. Our device consists of a Plexiglas system with a central channel and a gold electrode. For immobilization of the E. granulosus antigen, a gold electrode was modified with the incorporation of gold nanoparticles. Immobilized antigen was allowed to react with IgG-anti-E. granulosus antibodies in samples, and these were quantified by horseradish peroxidase (HRP) enzyme-labeled secondary antibodies specific to human IgG using catechol (Q) as enzymatic mediator. HRP in the presence of hydrogen peroxide (H(2)O(2)) catalyzes the oxidation of Q to o-benzoquinone (P). The electrochemical reduction back to Q was detected on the gold electrode (AuE) at -0.15 V. The current obtained was proportional to the activity of the enzyme and to the concentration of antibodies of interest. The detection limit for electrochemical detection was 0.091 ng ml(-1), and the within- and between-assay coefficients of variation were below 6.7%. The proposed system presents many benefits, the more relevant are: reduced complexity and costs that are considered as the most wanted features for the clinical-immunodiagnostic field.
本文描述了一种微流控免疫传感器,用于检测人血清样本中针对细粒棘球蚴的 IgG 抗体,这是一种替代工具,可用于自动免疫诊断包虫病。我们的设备由一个带有中央通道和一个金电极的有机玻璃系统组成。为了固定棘球蚴抗原,金电极通过掺入金纳米粒子进行修饰。固定化抗原与样品中的 IgG-抗棘球蚴抗体反应,并用辣根过氧化物酶(HRP)标记的针对人 IgG 的二抗进行定量,该二抗特异性针对 IgG-抗棘球蚴抗体,二抗使用儿茶酚(Q)作为酶介体。存在过氧化氢(H2O2)时,HRP 催化 Q 氧化为邻苯醌(P)。在 -0.15 V 下,在金电极(AuE)上检测到 Q 的电化学还原。获得的电流与酶的活性和感兴趣的抗体浓度成正比。电化学检测的检测限为 0.091ngml(-1),批内和批间变异系数均低于 6.7%。所提出的系统具有许多优点,其中更相关的是:降低了复杂性和成本,这被认为是临床免疫诊断领域最需要的特征。