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Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test

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Chikungunya Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1426))

Abstract

Neutralization assay is a technique that detects and quantifies neutralizing antibody in serum samples by calculating the percentage of reduction of virus activity, as the concentration of virus used is usually constant. Neutralizing antibody titer is conventionally determined by calculating the percentage reduction in total virus infectivity by counting and comparing number of plaques (localized area of infection due to cytopathic effect) with a standard amount of virus. Conventional neutralizing test uses plaque-reduction neutralization test (PRNT) to determine neutralizing antibody titers against Chikungunya virus (CHIKV). Here we describe the plaque reduction neutralization assay (PRNT) using Vero cell lines to obtain neutralizing antibody titers.

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Acknowledgements

We would like to acknowledge Dr. Chang-Kweng Lim, Department of Virology I, National Institute of Infectious Diseases, Japan, for generously providing us with a figure on CHIKV plaques (Fig. 3). This work was supported in part by the research grant, Research on Emerging and Re-emerging Infectious Diseases (H26-shinkou-jitsuyouka-007), from the Ministry of Health, Labour and Welfare, Japan, the Environment Research and Technology Development Fund (S-8) of the Ministry of the Environment, and a Grant-in-Aid for Young Scientists (B) from JSPS (26870872).

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Authors and Affiliations

  1. Department of Virology 1, National Institute of Infectious Diseases, Toyama 1-23-1 Shinjuku-ku, Tokyo, 162-8640, Japan

    Nor Azila Muhammad Azami, Meng Ling Moi & Tomohiko Takasaki

  2. Graduate School for Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577, Japan

    Nor Azila Muhammad Azami

  3. Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan

    Meng Ling Moi

Authors
  1. Nor Azila Muhammad Azami
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  2. Meng Ling Moi
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  3. Tomohiko Takasaki
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Corresponding author

Correspondence to Meng Ling Moi .

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Editors and Affiliations

  1. Dept of Microbiology and Immunology, National University of Singapore, Singapore, Singapore

    Justin Jang Hann Chu

  2. Dept of Microbiology and Immunology, National University of Singapore, Singapore, Singapore

    Swee Kim Ang

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Azami, N.A.M., Moi, M.L., Takasaki, T. (2016). Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test. In: Chu, J., Ang, S. (eds) Chikungunya Virus. Methods in Molecular Biology, vol 1426. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3618-2_25

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  • DOI: https://doi.org/10.1007/978-1-4939-3618-2_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3616-8

  • Online ISBN: 978-1-4939-3618-2

  • eBook Packages: Springer Protocols

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