Nguyen Binh T T, Koh Guiwan, Lim Hui Si, Chua Anthony J S, Ng Mary M L, Toh Chee-Seng
Department of Chemistry, Faculty of Science, Yong Loo Lin School of Medicine, 3 Science Drive 3, National University of Singapore, Singapore 117543.
Anal Chem. 2009 Sep 1;81(17):7226-34. doi: 10.1021/ac900761a.
A membrane-based electrochemical nanobiosensor sensitive toward whole viral particles is fabricated by forming a submicrometer thick nanoporous alumina membrane over a platinum disk electrode. Antibody probe molecules are physically adsorbed onto the walls of the membrane nanochannels. The sensing signal is based on the monitoring of the electrode's Faradaic current response toward ferrocenemethanol, which is extremely sensitive to the formation of immunocomplex within the nanoporous membrane. This nanobiosensor is demonstrated for the sensing of West Nile virus protein domain III (WNV-DIII) and the inactivated West Nile viral particle, using anti-WNV-DIII immunoglobulin M (IgM) as the biorecognition probe. The detection of the viral protein and the particle are logarithmically linear up to 53 pg mL(-1) (R(2) = 0.99) and 50 viral particles per 100 mL (R(2) = 0.93) in pH 7, with extremely low detection limits of 4 pg mL(-1) and ca. 2 viral particles per 100 mL, comparable to sensitivities of polymerase chain reaction techniques. The relative standard deviation (RSD) of whole viral particle detection in whole blood serum is 6.9%. In addition, the simple nanobiosensor construction procedure, minimal sample preparation, and short detection time of 30 min are highly attractive properties and demonstrate that the detection of a wide range of proteins and viruses can be achieved.
通过在铂盘电极上形成亚微米厚的纳米多孔氧化铝膜,制备了一种对完整病毒颗粒敏感的基于膜的电化学纳米生物传感器。抗体探针分子物理吸附在膜纳米通道的壁上。传感信号基于监测电极对二茂铁甲醇的法拉第电流响应,该响应对纳米多孔膜内免疫复合物的形成极其敏感。使用抗西尼罗河病毒DIII结构域免疫球蛋白M(IgM)作为生物识别探针,展示了这种纳米生物传感器用于检测西尼罗河病毒蛋白结构域III(WNV-DIII)和灭活的西尼罗河病毒颗粒。在pH 7条件下,病毒蛋白和病毒颗粒的检测在高达53 pg mL⁻¹(R² = 0.99)和每100 mL 50个病毒颗粒(R² = 0.93)时呈对数线性,检测限极低,分别为4 pg mL⁻¹和约每100 mL 2个病毒颗粒,与聚合酶链反应技术的灵敏度相当。全血血清中完整病毒颗粒检测的相对标准偏差(RSD)为6.9%。此外,简单的纳米生物传感器构建程序、最少的样品制备以及30分钟的短检测时间具有高度吸引力,表明可以实现对多种蛋白质和病毒的检测。