title: Detection of avian influenza virus by fluorescent DNA barcode-based immunoassay with sensitivity comparable to PCR.
abstract: In this paper, a coupling of fluorophore-DNA barcode and bead-based immunoassay for detecting avian influenza virus (AIV) with PCR-like sensitivity is reported. The assay is based on the use of sandwich immunoassay and fluorophore-tagged oligonucleotides as representative barcodes. The detection involves the sandwiching of the target AIV between magnetic immunoprobes and barcode-carrying immunoprobes. Because each barcode-carrying immunoprobe is functionalized with a multitude of fluorophore-DNA barcode strands, many DNA barcodes are released for each positive binding event resulting in amplification of the signal. Using an inactivated H16N3 AIV as a model, a linear response over five orders of magnitude was obtained, and the sensitivity of the detection was comparable to conventional RT-PCR. Moreover, the entire detection required less than 2 hr. The results indicate that the method has great potential as an alternative for surveillance of epidemic outbreaks caused by AIV, other viruses and microorganisms.
authors.affiliation: DTU-Nanotech, Department of Micro and Nanotechnology, Technical University of Denmark, DTU Building 345 East, DK-2800 Kongens Lyngby, Denmark.
authors.forename: Cuong
authors.initials: C
authors.lastname: Cao
authors.forename: Raghuram
authors.initials: R
authors.lastname: Dhumpa
authors.forename: Dang Duong
authors.initials: DD
authors.lastname: Bang
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Ghavifekr
authors.forename: Jonas
authors.initials: J
authors.lastname: Høgberg
authors.forename: Anders
authors.initials: A
authors.lastname: Wolff
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D002645:Chickens; D007118:Immunoassay; D009980:Influenza A virus / Q000235:genetics* / Q000472:pathogenicity; D005585:Influenza in Birds / Q000175:diagnosis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013050:Spectrometry, Fluorescence