Abstract

Discrimination of closely related strains is a key issue, particularly for infectious diseases whose incidence fluctuates according to variations in the season and evolutionary changes. Among infectious diseases, influenza viral infections are a worldwide cause of pandemic disease and mortality. With the emergence of different influenza strains, it is vital to develop a method using antibodies that can differentiate between viral types and subtypes. Ideally, such a system would also be user friendly. In this study, a polyclonal antibody generated against A/Udorn/307/1972 (H3N2) was used as a probe to distinguish between influenza H3N2 viruses based on the interaction between the antibody and hemagglutinin, demonstrating its applicability for viral discrimination. Clear discrimination was demonstrated using an evanescent-field-coupled waveguide-mode sensor, which has appealing characteristics over other methods in the viewpoint of improving the sensitivity, measurement time, portability and usability. Further supporting evidence was obtained using enzyme-linked immunosorbent assays, hemagglutination-inhibition assays, and infectivity neutralization assays. The results obtained indicate that the polyclonal antibody used here is a potential probe for distinguishing influenza viruses and, with the aid of a handheld sensor it could be used for influenza surveillance.

Highlights

  • Influenza viruses are spherical or filamentous, enveloped, and range in size from 80 to 100 nm [1]

  • We have previously demonstrated the antibody-based detection of human and avian influenza viruses using the evanescentfield-coupled waveguide-mode (EFC-WM) sensor and the detection limit of this sensor was determined to be in the order of 103 pfu/ml [6,23]

  • Different anti-HA probes are available for detection of viral infections; these have been implemented in several diagnostic methods, including enzyme-linked immunosorbent assays, immunoblots, immunosensor-based methods, interferometry, fluoroimmunoassays, methods based on surface plasmon resonance (SPR), and immunochromatography [6,7,8,9,10,11,26,28]

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Summary

Introduction

Influenza viruses are spherical or filamentous, enveloped, and range in size from 80 to 100 nm [1]. To pinpoint differences among influenza H3N2 viruses by using a polyclonal antibody as the probe, we developed a detection method using an anti-A/Udorn/307/1972 polyclonal antibody on an evanescentfield-coupled waveguide-mode (EFC-WM) sensor. The present study explores a further step, using antibody-based sensing for discrimination of influenza viruses belonging to the H3N2 subtype.

Results
Conclusion
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