Diagnostic Virology Protocols pp 17-38

Part of the Methods in Molecular Biology book series (MIMB, volume 665)


  • A. M. Powers
  • John T. Roehrig


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.

Key words

Alphavirus Nucleic acid amplification test ELISA Monoclonal antibodies 


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • A. M. Powers
    • 1
  • John T. Roehrig
    • 1
  1. 1.Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, U.S. Department of Health and Human Services, Centers for Disease Control and PreventionPublic Health ServiceFort CollinsUSA

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