A virus-type specific serological diagnosis of flavivirus infection using virus-like particles

Min Qing, Zhi Ming Yuan, Pei Yong Shi

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus (WNV), dengue virus (DENV), yellow fever virus (YFV), and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test (PRNT) is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety level-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles (VLPs) that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps: (i) VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37°C for 1 h; (ii)the neutralized VLPs are used to infect Vero cells; and (iii) the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen (as quantified by the luciferase activities) is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the "gold standard" of the classic PRNT; importantly, it shortens the assay time from >10 days to <1 day, and can be performed in biosafety level-2 facility.

Original languageEnglish (US)
Pages (from-to)136-145
Number of pages10
JournalVirologica Sinica
Volume24
Issue number2
DOIs
StatePublished - Apr 29 2009
Externally publishedYes

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Keywords

  • Flavivirus packaging
  • Neutralization assay
  • Serological diagnosis
  • Viral-like particle
  • West Nile virus

ASJC Scopus subject areas

  • Immunology
  • Molecular Medicine
  • Virology

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