Development of Neutralization Assay Using an eGFP Chikungunya Virus

Viruses. 2016 Jun 28;8(7):181. doi: 10.3390/v8070181.

Abstract

Chikungunya virus (CHIKV), a member of the Alphavirus genus, is an important human emerging/re-emerging pathogen. Currently, there are no effective antiviral drugs or vaccines against CHIKV infection. Herein, we construct an infectious clone of CHIKV and an eGFP reporter CHIKV (eGFP-CHIKV) with an isolated strain (assigned to Asian lineage) from CHIKV-infected patients. The eGFP-CHIKV reporter virus allows for direct visualization of viral replication through the levels of eGFP expression. Using a known CHIKV inhibitor, ribavirin, we confirmed that the eGFP-CHIKV reporter virus could be used to identify inhibitors against CHIKV. Importantly, we developed a novel and reliable eGFP-CHIKV reporter virus-based neutralization assay that could be used for rapid screening neutralizing antibodies against CHIKV.

Keywords: Chikungunya virus; antiviral; neutralization antibody; reporter virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / immunology*
  • Antibodies, Viral / immunology*
  • Cell Line
  • Chikungunya virus / genetics
  • Chikungunya virus / immunology*
  • Chikungunya virus / physiology
  • Genes, Reporter*
  • Green Fluorescent Proteins / analysis*
  • Green Fluorescent Proteins / genetics
  • Mass Screening / methods
  • Neutralization Tests / methods*
  • Recombinant Proteins / analysis
  • Recombinant Proteins / genetics
  • Staining and Labeling / methods*
  • Virus Replication

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Recombinant Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins