Efficient capture of infectious H5 avian influenza virus utilizing magnetic beads coated with anionic polymer
Akikazu Sakudo1, Kazuyoshi Ikuta
1Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan. sakudo@biken.osaka-u.ac.jp
Abstract:
The possible emergence of a pandemic influenza virus from the avian influenza virus (AIV) has become a serious threat. The isolation of viruses will be crucial for further virological analysis and the development of vaccines. However, currently, there is no simple method for facilitating the isolation of infectious AIV. Here, we have developed a simple method of capturing AIV using anionic magnetic beads. The method employed the capture of AIV (H5N1, H5N2, and H5N3) from liquid samples such as allantoic fluid (AF) and cell culture medium (CM) using magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic anhydride). After their incubation with AIV-containing samples, the magnetic beads were separated from the supernatant by applying a magnetic field. The absorption of AIV on the beads was confirmed by immunochromatography and an enzyme-linked immunosorbent assay, which indicated the presence of hemagglutinin, neuraminidase, and nucleoprotein of AIV. Furthermore, the infectivity in chicken eggs of AIV captured by magnetic beads was similar to that of the starting materials. The capture of AIV using magnetic beads coated with anionic polymers will contribute to the sufficient recovery of infectious AIV and approach for potential pandemic influenza viruses.
Insights
A new method uses anionic magnetic beads to easily capture infectious avian influenza viruses (AIV) from samples. This technique aids in isolating AIV for research and developing vaccines against potential pandemics.
Area of Science:
- Virology
- Biotechnology
- Infectious Diseases
Background:
- Avian influenza virus (AIV) poses a pandemic threat.
- Efficient isolation of infectious AIV is critical for virological analysis and vaccine development.
- Current methods for AIV isolation are not simple or efficient.
Purpose of the Study:
- To develop a simple and effective method for capturing infectious AIV.
- To utilize anionic magnetic beads for AIV isolation from various liquid samples.
- To facilitate the study of AIV and the development of pandemic influenza countermeasures.
Main Methods:
- Developed a method using anionic magnetic beads coated with poly(methyl vinyl ether-maleic anhydride).
- Captured AIV (H5N1, H5N2, H5N3) from allantoic fluid and cell culture medium.
- Separated magnetic beads using a magnetic field and confirmed AIV capture via immunochromatography and ELISA.
Main Results:
- Successfully captured AIV from liquid samples using anionic magnetic beads.
- Confirmed the presence of AIV hemagglutinin, neuraminidase, and nucleoprotein on the captured beads.
- Demonstrated that the infectivity of captured AIV in chicken eggs was comparable to the original samples.
Conclusions:
- Anionic magnetic beads provide a simple and efficient method for capturing infectious AIV.
- This technique facilitates the recovery of viable AIV for further research.
- The method offers a promising approach for studying and controlling potential pandemic influenza viruses.
More Related Videos
08:10Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses
Published on: January 15, 2020
08:10A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example
Published on: May 29, 2012