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Updated: Sep 24, 2025
A Miniaturized Glycan Microarray Assay for Assessing Avidity and Specificity of Influenza A Virus Hemagglutinins
Published on: May 29, 2016
Detection of Influenza Virus by Agglutination of Microparticles Immobilized a Mixed Glycan Receptor Produced from
Teruhiko Matsubara1, Ayaka Ogami1, Haruka Kori1
1Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama 223-8522, Japan.
Abstract:
The hemagglutination inhibition (HAI) assay is one of the detection methods for influenza virus (IFV) under global influenza surveillance, which uses freshly prepared animal red blood cells (RBCs). Here, we demonstrate that a mixed glycan-modified polystyrene microparticle, which can be chemically prepared in advance, can replace animal RBCs in the HAI assay. A mixture of azide-conjugated glycans containing sialyl- and sulfated-lactose moieties was produced from Madin-Darby canine kidney (MDCK) cells, which are used for IFV isolation, and then immobilized on the surface of a polystyrene microparticle using click chemistry. Human HA and IFV were detected with high sensitivity when using the mixed glycan-immobilized particle.
Insights
This study introduces a novel synthetic microparticle for influenza virus detection, replacing animal red blood cells in hemagglutination inhibition assays. This innovation offers a stable, pre-prepared alternative for global influenza surveillance.
Area of Science:
- Virology and Immunology
- Biotechnology and Nanotechnology
- Diagnostic Assay Development
Background:
- The hemagglutination inhibition (HAI) assay is crucial for influenza virus (IFV) detection in global surveillance.
- Traditional HAI assays rely on freshly prepared animal red blood cells (RBCs), posing logistical and consistency challenges.
- There is a need for stable, readily available reagents to enhance influenza diagnostic capabilities.
Purpose of the Study:
- To develop and validate a synthetic alternative to animal RBCs for use in HAI assays.
- To demonstrate the efficacy of glycan-modified microparticles in detecting human hemagglutinin (HA) and IFV.
- To provide a more stable and accessible reagent for influenza virus surveillance.
Main Methods:
- Chemically synthesized azide-conjugated glycans, including sialyl- and sulfated-lactose moieties, derived from MDCK cells.
- Immobilization of these mixed glycans onto polystyrene microparticles via click chemistry.
- Utilized the glycan-immobilized microparticles in a modified HAI assay for influenza detection.
Main Results:
- Successfully developed mixed glycan-modified polystyrene microparticles as a replacement for animal RBCs.
- Demonstrated high sensitivity in detecting both human HA and infectious influenza virus using the microparticle-based assay.
- The synthetic particles offer a stable, pre-prepared reagent suitable for advance preparation.
Conclusions:
- Mixed glycan-modified microparticles are a viable and sensitive alternative to animal RBCs in HAI assays.
- This approach offers significant advantages for influenza virus (IFV) detection and global surveillance.
- The chemically prepared microparticles enhance the stability and accessibility of diagnostic reagents.
