title: Rapid detection of avian influenza virus H5N1 in chicken tracheal samples using an impedance aptasensor with gold nanoparticles for signal amplification.
abstract: Highly pathogenic avian influenza virus H5N1 is a continuous threat to public health and poultry industry. The recurrence of the H5N1 led us to develop a robust, specific, and rapid detection method for the virus. In this study, an impedance aptasensor was developed for the virus detection using specific H5N1 aptamer and a gold interdigitated microelectrode. Streptavidin was immobilized on the microelectrode surface and biotin labeled H5N1 aptamer was bound to the immobilized streptavidin. The microelectrode was blocked with the polyethylene glycol and the bound aptamer captured the virus. The impedance change caused by the captured virus was measured using an impedance analyzer. To enhance impedance signal, a nanoparticle-based amplifier was designed and implemented by forming a network-like gold nanoparticles/H5N1-aptamer/thiocyanuric acid. The detection limit of the impedance aptasensor was 0.25 HAU for the pure virus and 1 HAU for the tracheal chicken swab samples spiked with the H5N1 virus. The detection time of aptasensor without employing the amplifier was less than an hour. The amplifier increased impedance by a 57-fold for the 1 HAU samples. Only negligible impedance change was observed for non-target viruses such as H5N2, H5N3, H7N2, H1N1, and H2N2. This aptasensor provides a foundation for the development of a portable aptasensor instrument.
authors.affiliation: Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Sardar
authors.initials: S
authors.lastname: Karash
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Ronghui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Kelso
authors.affiliation: Animal Diagnostic Laboratory, State University, State College, PA 16802, USA.
authors.forename: Huaguang
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Engineering Science and Mechanics & Material Research Institute, Pennsylvania State University, State College, PA 16802, USA.
authors.forename: Tony Jun
authors.initials: TJ
authors.lastname: Huang
authors.affiliation: Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA; Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA; Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. Electronic address: yanbinli@uark.edu.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D002645:Chickens; D017097:Electric Impedance; D053124:Influenza A Virus, H5N1 Subtype / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D008839:Microelectrodes; D053758:Nanoparticles / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014132:Trachea / Q000821:virology*
chemical_list: D052157:Aptamers, Nucleotide
keywords: Amplifier; Aptamer; Aptasensor; Avian influenza H5N1; Microelectrode