Abstract
Alphaviruses remain important emerging mosquito-borne, zoonotic pathogens that cause both localized human outbreaks and epizootics (e.g., Venezuelan equine encephalitis) and large human epidemics (e.g., Chikungunya). Alphaviruses are globally dispersed, and each continent has humans at risk from one or more of these arthropod-borne viruses (arboviruses). Symptoms of human alphaviral disease range from frank, severe encephalitis (e.g., eastern and western equine encephalitis) to polyarthritis (e.g., Ross River). Diagnostic techniques to identify human alphaviral infections have changed dramatically with the development and implementation of standardized nucleic acid amplification tests (NAAT). The NAAT is rapidly replacing virus isolation and typing using indirect fluorescent antibody (IFA) assay with monoclonal antibodies (MAbs) as the preferred method of virus identification. The older techniques still have value, however, since alphaviral growth in cell culture is rapid, and IFA with MAbs is inexpensive. This chapter provides detailed, standardized protocols for the identification of alphaviruses from clinical specimens and the serological characterization of human infection-immune sera. Both laboratory approaches are needed to identify and confirm human infections with these agents.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Roehrig, J.T. (1986) The use of monoclonal antibodies in studies of the structural proteins of alphaviruses and flaviviruses. In: Schlesinger, S. and Schlesinger, M.J. Ed. The Viruses: The Togaviridae and Flaviviridae. Plenum Press, New York, pp. 251–278.
Powers, A.M., Brault, A.C., Kang, W., and Weaver, S.C. (2001) Evolutionary relationships and systematics of the alphaviruses. J Virol 75, 10119–10131.
Tsai, T.H. (1992) Arboviruses. In: Rose, N.R., Marcario, E.C., Fahey, J.L., Friedman, H., and Penn, G.M. Ed. Manual of Clinical Laboratory Immunology, 4th Edn. American Society for Microbiology, Washington, DC, pp. 606–618.
Voller, A., Bidwell, D., and Bartlett, A. (1976) Microplate immunoassay for the immunodiagnosis of virus infections. In: Rose, N.R. and Friedman, H.H. Ed. Handbook of Clinical Immunology. American Society for Microbiology, Washington, DC, pp. 506–512.
Powers, A.M., Aguilar, P.V., Chandler, L.J., Brault, A.C., Meakins, T.A., Watts, D.M., Vasconcelos, P.F.C., Travassos Da Rosa, A.P., Weaver, S.C., and Tesh, R.B. (2006) Genetic relationships of Mayaro and Una viruses suggests distinct patterns of transmission. Am J Trop Med Hyg 75(3), 461–469.
Kariuki Njenga, M., Nderitu, L., Ledermann, J.P., Ndirangu, A., Logue, C.H., Kelly, C.H.L., Sang, R., Sergon, K., Breiman, R., and Powers, A.M. (2008) Tracking epidemic chikungunya virus into the Indian Ocean from East Africa. J Gen Virol 89, 2754–2760.
Roehrig, J.T., Hunt, A.R., Chang, G.-J., Sheik, B., Bolin, R.A., Tsai, T.F., and Trent, D.W. (1990) Identification of monoclonal antibodies capable of differentiating antigenic varieties of eastern equine encephalitis viruses. Am J Trop Med Hyg 42, 394–398.
Hunt, A.R. and Roehrig, J.T. (1985) Biochemical and biological characteristics of epitopes on the E1 glycoprotein of western equine encephalitis virus. Virology 142, 334–344.
Tsai, T.F., Bolin, R.A., Montoya, M., Bailey, R.E., Francy, D.B., Jozan, M., and Roehrig, J.T. (1987) Detection of St. Louis encephalitis virus antigen in mosquitoes by capture enzyme immunoassay. J Clin Microbiol 25, 370–376.
Roehrig, J.T., Bolin, R.A., Hunt, A.R., and Woodward, T.M. (1991) Use of a new synthetic peptide derived monoclonal antibody to differentiate vaccine from wild-type Venezuelan equine encephalomyelitis viruses. J Clin Microbiol 29, 630–631.
Roehrig, J.T., Day, J.W., and Kinney, R.M. (1982) Antigenic analysis of the surface glycoproteins of a Venezuelan equine encephalomyelitis virus (TC-83) using monoclonal antibodies. Virology 118, 269–278.
Roehrig, J.T. and Mathews, J.H. (1985) The neutralization site on the E2 glycoprotein of Venezuelan equine encephalomyelitis (TC-83) virus is composed of multiple conformationally stable epitopes. Virology 142, 347–356.
Rico-Hesse, R., Roehrig, J.T., and Dickerman, R.W. (1988) Monoclonal antibodies define antigenic variation within the ID variety of Venezuelan equine encephalitis virus. Am J Trop Med Hyg 38, 187–194.
Karabatsos, N., Lewis, A.L., Calisher, C.H., Hunt, A.R., and Roehrig, J.T. (1988) Identification of Highlands J virus from a Florida horse. Am J Trop Med Hyg 39, 603–606.
Schmaljohn, A.L., Johnson, E.D., Dalrymple, J.M., Cole, G.A. (1983) Non-neutralizing monoclonal antibodies can prevent lethal alphavirus encephalitis. Nature 297, 70–72.
Boere, W.A.M., Harnsen, M., Vinje, J., Benaissa-Trouw, B.J., Kraaijeveld, C.A., and Snippe, H. (1984) Identification of distinct determinants on Semliki Forest virus by using monoclonal antibodies with different antiviral activities. J Virol 52, 575–582.
Wulff, H. and Lange, J.V. (1975) Indirect immunofluorescence for the diagnosis of Lassa fever infection. Bull WHO 52, 429–436.
Wulff, H., Lange, J.V., and Webb, P.A. (1978) Interrelationships among arenaviruses measured by indirect immunofluorescence. Intervirology 9, 344–350.
Riggs, J.L. (1979) Immunofluorescent staining. In: Lennette, E.H. and Schmidt, N.J. Ed. Diagnostic Procedures for Viral, Rickettsia, and Chlamydial Infections, 5th Edn. American Public Health Association, Washington, DC.
Acknowledgements
The authors would like to thank Teresa Brown, Alison Johnson, Denise Martin, Nick Karabatsos, Rob Lanciotti, Roger Nasci, and Jeremy Ledermann for their contributions to the development and standardization of these protocols. Disclaimer. “The findings and conclusions in this report are those of the author(s) and do not necessarily represent the views of [the Centers for Disease Control and Prevention/the Agency for Toxic Substances and Disease Registry].”
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Powers, A.M., Roehrig, J.T. (2010). Alphaviruses. In: Stephenson, J., Warnes, A. (eds) Diagnostic Virology Protocols. Methods in Molecular Biology, vol 665. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-817-1_2
Download citation
DOI: https://doi.org/10.1007/978-1-60761-817-1_2
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-816-4
Online ISBN: 978-1-60761-817-1
eBook Packages: Springer Protocols