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Updated: Nov 4, 2025
High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
Development of 6E3 antibody-mediated SERS immunoassay for drug-resistant influenza virus
Hyeran Kim1, Hyunju Kang1, Hye-Nan Kim1
1Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
Abstract:
Influenza viruses are responsible for several pandemics and seasonal epidemics and pose a major public health threat. Even after a major outbreak, the emergence of drug-resistant influenza viruses can pose disease control problems. Here we report a novel 6E3 monoclonal antibody capable of recognizing and binding to the H275Y neuraminidase (NA) mutation, which has been associated with reduced susceptibility of influenza viruses to NA inhibitors. The 6E3 antibody had a KD of 72.74 μM for wild-type NA and 32.76 pM for H275Y NA, suggesting that it can identify drug-resistant pandemic H1N1 (pH1N1) influenza virus. Molecular modeling studies also suggest the high-affinity binding of this antibody to pH1N1 H275Y NA. This antibody was also subject to dot-blot, enzyme-linked immunosorbent assay, bare-eye detection, and lateral flow assay to demonstrate its specificity to drug-resistant pH1N1. Furthermore, it was immobilized on Au nanoplate and nanoparticles, enabling surface-enhanced Raman scattering (SERS)-based detection of the H275Y mutant pH1N1. Using 6E3 antibody-mediated SERS immunoassay, the drug-resistant influenza virus can be detected at a low concentration of 102 plaque-forming units/mL. We also detected pH1N1 in human nasopharyngeal aspirate samples, suggesting that the 6E3-mediated SERS assay has the potential for diagnostic application. We anticipate that this newly developed antibody and SERS-based immunoassay will contribute to the diagnosis of drug-resistant influenza viruses and improve treatment strategies for influenza patients.
Insights
A new 6E3 monoclonal antibody specifically detects drug-resistant pandemic H1N1 influenza virus with a H275Y neuraminidase mutation. This antibody enables rapid, sensitive detection, aiding in improved influenza diagnosis and treatment strategies.
Area of Science:
- Virology
- Immunology
- Nanotechnology
Background:
- Influenza viruses cause pandemics and seasonal epidemics, posing significant public health risks.
- Emergence of drug-resistant strains complicates disease control efforts.
- Neuraminidase (NA) inhibitors are key antiviral drugs, but resistance mutations like H275Y reduce their efficacy.
Purpose of the Study:
- To develop a novel monoclonal antibody for specific detection of drug-resistant influenza viruses.
- To characterize the binding affinity of the antibody to wild-type and mutant neuraminidase.
- To establish a sensitive and rapid diagnostic assay for drug-resistant pandemic H1N1 (pH1N1).
Main Methods:
- Development and characterization of the 6E3 monoclonal antibody.
- Binding affinity assays (KD measurements) for wild-type and H275Y NA.
- Molecular modeling for binding site analysis.
- Dot-blot, ELISA, and lateral flow assays for specificity testing.
- Immobilization of antibody on gold nanoparticles for SERS-based detection.
- Detection of pH1N1 in clinical samples (nasopharyngeal aspirates).
Main Results:
- The 6E3 antibody exhibits high affinity for the H275Y NA mutation (32.76 pM) compared to wild-type NA (72.74 μM).
- Demonstrated specificity for drug-resistant pH1N1 via multiple immunoassay formats.
- Developed a SERS-based immunoassay capable of detecting drug-resistant influenza virus at concentrations as low as 102 PFU/mL.
- Successfully detected pH1N1 in human nasopharyngeal aspirate samples.
Conclusions:
- The 6E3 monoclonal antibody is a promising tool for identifying drug-resistant pH1N1 influenza.
- The developed SERS immunoassay offers a sensitive and rapid method for detecting resistant strains.
- This technology has potential for clinical diagnostic applications, aiding in timely and effective influenza treatment.
