title: An Impedance Aptasensor with Microfluidic Chips for Specific Detection of H5N1 Avian Influenza Virus.
abstract: 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.
authors.affiliation: Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA. jlum@pacificvetgroup.com.
authors.forename: Jacob
authors.initials: J
authors.lastname: Lum
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA. rwang@uark.edu.
authors.forename: Ronghui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. bhargis@uark.edu.
authors.forename: Billy
authors.initials: B
authors.lastname: Hargis
authors.affiliation: Department of Mechanical Engineering, University of Arkansas, Fayetteville, AR 72701, USA. chstung@uark.edu.
authors.forename: Steve
authors.initials: S
authors.lastname: Tung
authors.affiliation: Department of Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. wbottje@uark.edu.
authors.forename: Walter
authors.initials: W
authors.lastname: Bottje
authors.affiliation: Animal Diagnostic Laboratory, Pennsylvania State University, University Park, PA 16802, USA. hxl15@psu.edu.
authors.forename: Huaguang
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA. yanbinli@uark.edu.|Department of Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. yanbinli@uark.edu.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D001717:Birds / Q000821:virology; D017097:Electric Impedance; D053124:Influenza A Virus, H5N1 Subtype / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000821:virology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D044085:Microfluidics / Q000295:instrumentation*
chemical_list: D052157:Aptamers, Nucleotide
keywords: aptamer; avian influenza virus; impedance aptasensor; microfluidic chip; virus detection