An Impedance Aptasensor with Microfluidic Chips for Specific Detection of H5N1 Avian Influenza Virus

Sensors

Jacob Lum⁴  Ronghui Wang³  Billy Hargis²  Steve Tung⁵  Walter Bottje²  Huaguang Lu¹  Yanbin Li³ 

[1]Animal Diagnostic Laboratory, Pennsylvania State University, University Park, PA 16802, USA

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[3]Department of Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA

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[5]Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA

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[7]Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA

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[9]Department of Mechanical Engineering, University of Arkansas, Fayetteville, AR 72701, USA

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关键词: impedance aptasensor;    avian influenza virus;    aptamer;    microfluidic chip;    virus detection;   

DOI  :  10.3390/s150818565

来源: mdpi

【 摘 要 】

In this research a DNA aptamer, which was selected through SELEX (systematic evolution of ligands by exponential enrichment) to be specific against the H5N1 subtype of the avian influenza virus (AIV), was used as an alternative reagent to monoclonal antibodies in an impedance biosensor utilizing a microfluidics flow cell and an interdigitated microelectrode for the specific detection of H5N1 AIV. The gold surface of the interdigitated microelectrode embedded in a microfluidics flow cell was modified using streptavidin. The biotinylated aptamer against H5N1 was then immobilized on the electrode surface using biotin–streptavidin binding. The target virus was captured on the microelectrode surface, causing an increase in impedance magnitude. The aptasensor had a detection time of 30 min with a detection limit of 0.0128 hemagglutinin units (HAU). Scanning electron microscopy confirmed the binding of the target virus onto the electrode surface. The DNA aptamer was specific to H5N1 and had no cross-reaction to other subtypes of AIV (e.g., H1N1, H2N2, H7N2). The newly developed aptasensor offers a portable, rapid, low-cost alternative to current methods with the same sensitivity and specificity.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.