Assessment of plaque assay methods for alphaviruses

Diana Juarez, Kanya C. Long, Patricia Aguilar, Tadeusz J. Kochel, Eric S. Halsey

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Viruses from the Alphavirus genus are responsible for numerous arboviral diseases impacting human health throughout the world. Confirmation of acute alphavirus infection is based on viral isolation, identification of viral RNA, or a fourfold or greater increase in antibody titers between acute and convalescent samples. In convalescence, the specificity of antibodies to an alphavirus may be confirmed by plaque reduction neutralization test. To identify the best method for alphavirus and neutralizing antibody recognition, the standard solid method using a cell monolayer overlay with 0.4% agarose and the semisolid method using a cell suspension overlay with 0.6% carboxymethyl cellulose (CMC) overlay were evaluated. Mayaro virus, Una virus, Venezuelan equine encephalitis virus (VEEV), and Western equine encephalitis virus (WEEV) were selected to be tested by both methods. The results indicate that the solid method showed consistently greater sensitivity than the semisolid method. Also, a " semisolid-variant method" using a 0.6% CMC overlay on a cell monolayer was assayed for virus titration. This method provided the same sensitivity as the solid method for VEEV and also had greater sensitivity for WEEV titration. Modifications in plaque assay conditions affect significantly results and therefore evaluation of the performance of each new assay is needed.

Original languageEnglish (US)
Pages (from-to)185-189
Number of pages5
JournalJournal of Virological Methods
Volume187
Issue number1
DOIs
StatePublished - Jan 1 2013

Keywords

  • Agarose
  • Alphavirus
  • Carboxymethylcellulose
  • Plaque assay

ASJC Scopus subject areas

  • Virology

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