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Updated: Jul 16, 2026
High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
Rapid Diagnostic Assay for Intact Influenza Virus Using a High Affinity Hemagglutinin Binding Protein
Caitlin E Anderson1, Carly A Holstein1, Eva-Maria Strauch2
1Department of Bioengineering, University of Washington , Seattle, Washington 98195-5061, United States.
Abstract:
Influenza is a ubiquitous and recurring infection that results in approximately 500 000 deaths globally each year. Commercially available rapid diagnostic tests are based upon detection of the influenza nucleoprotein, which are limited in that they are unable to differentiate by species and require an additional viral lysis step. Sample preprocessing can be minimized or eliminated by targeting the intact influenza virus, thereby reducing assay complexity and leveraging the large number of hemagglutinin proteins on the surface of each virus. Here, we report the development of a paper-based influenza assay that targets the hemagglutinin protein; the assay employs a combination of antibodies and novel computationally designed, recombinant affinity proteins as the capture and detection agents. This system leverages the customizability of recombinant protein design to target the conserved receptor-binding pocket of the hemagglutinin protein and to match the trimeric nature of hemagglutinin for improved avidity. Using this assay, we demonstrate the first instance of intact influenza virus detection using a combination of antibody and affinity proteins within a porous network. The recombinant head region binder based assays yield superior analytical sensitivity as compared to the antibody based assay, with lower limits of detection of 3.54 × 107 and 1.34 × 107 CEID50/mL for the mixed and all binder stacks, respectively. Not only does this work describe the development of a novel influenza assay, it also demonstrates the power of recombinant affinity proteins for use in rapid diagnostic assays.
Insights
A new paper-based influenza diagnostic assay targets the intact virus hemagglutinin protein. This novel approach uses recombinant affinity proteins and antibodies for sensitive detection, improving upon existing rapid influenza diagnostic tests.
Area of Science:
- Biotechnology
- Infectious Disease Diagnostics
- Protein Engineering
Background:
- Influenza causes significant global mortality annually.
- Current rapid diagnostic tests for influenza have limitations, including inability to differentiate species and requiring viral lysis.
- Targeting intact influenza virus, specifically the hemagglutinin protein, can simplify diagnostics and improve efficiency.
Purpose of the Study:
- To develop a novel paper-based assay for detecting intact influenza virus.
- To utilize a combination of antibodies and computationally designed recombinant affinity proteins.
- To enhance diagnostic sensitivity and specificity by targeting the hemagglutinin protein's receptor-binding pocket.
Main Methods:
- Development of a paper-based assay utilizing a combination of antibodies and recombinant affinity proteins.
- Design of recombinant proteins targeting the conserved receptor-binding pocket of the influenza hemagglutinin protein.
- Detection of intact influenza virus within a porous network using the developed assay.
Main Results:
- Demonstrated the first instance of intact influenza virus detection using antibody and recombinant affinity proteins in a porous network.
- Recombinant protein-based assays showed superior analytical sensitivity compared to antibody-only assays.
- Achieved lower limits of detection (1.34 × 10^7 CEID50/mL) with optimized binder combinations.
Conclusions:
- The developed paper-based assay offers a novel method for rapid influenza diagnostics.
- Recombinant affinity proteins are powerful tools for enhancing the sensitivity and specificity of diagnostic assays.
- This approach minimizes sample preprocessing and improves upon existing influenza diagnostic technologies.
