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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Staples JE, Breiman RF, Powers AM (2009) Chikungunya fever: an epidemiological review of a re-emerging infectious disease. Clin Infect Dis 49(6):942–948
Mallilankaraman K, Shedlock DJ, Bao H, Kawalekar OU, Fagone P, Ramanathan AA, Ferraro B, Stabenow J, Vijayachari P, Sundaram SG, Muruganandam N, Sarangan G, Srikanth P, Khan AS, Lewis MG, Kim JJ, Sardesai NY, Muthumani K, Weiner DB (2011) A DNA vaccine against Chikungunya virus is protective in mice and induces neutralizing antibodies in mice and nonhuman primates. PLoS Negl Trop Dis 5(1):e928. doi:10.1371/journal.pntd.0000928
Roehrig J, Hombach J, Barrett A (2008) Guidelines for plaque-reduction neutralization testing of human antibodies to dengue viruses. Viral Immunol 21:123–132
Thomas SJ, Nisalak A, Anderson KB, Libraty DH, Kalayanarooj S, Vaughn DW, Putnak R, Gibbons RV, Jarman R, Endy TP (2009) Dengue Plaque Reduction Neutralization Test (PRNT) in primary and secondary dengue virus infections: how alterations in assay conditions impact performance. Am J Trop Med Hyg 81(5):825–833. doi:10.4269/ajtmh.2009.08-0625
Dulbecco R, Vogt M, Strickland AGR (1956) A study of the basic aspects of neutralization of two animal viruses, Western equine encephalitis virus and poliomyelitis virus. Virology 2(2):162–205
Russell PK, Nisalak A, Sukhavachana P, Vivona S (1967) A Plaque Reduction Test for dengue virus neutralizing antibodies. J Immunol 99(2):285–290
Hobson-Peters J (2012) Approaches for the development of rapid serological assays for surveillance and diagnosis of infections caused by zoonotic flaviviruses of the Japanese encephalitis virus serocomplex. BioMed Res Int 2012:379738
Walker T, Jeffries CL, Mansfield KL, Johnson N (2014) Mosquito cell lines: history, isolation, availability and application to assess the threat of arboviral transmission in the United Kingdom. Parasit Vectors 7(1):382. doi:10.1186/1756-3305-7-382
Stim TB (1969) Arbovirus plaquing in two Simian kidney cell lines. J Gen Virol 5(3):329–338. doi:10.1099/0022-1317-5-3-329
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).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3618-2_25
Published:
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