Construction of eGFP-Tagged Senecavirus A for Facilitating Virus Neutralization Test and Antiviral Assay

Viruses. 2020 Mar 5;12(3):283. doi: 10.3390/v12030283.

Abstract

Senecavirus A (SVA), also known as Seneca Valley virus, is an emerging virus that causes vesicular disease in pigs. This virus belongs to the genus Senecavirus in the family Picornaviridae. The SVA CH-LX-01-2016 was isolated from Guangdong Province of China in 2016. In this study, a recombinant SVA CH-LX-01-2016 was constructed using reverse genetics, and proven to be able to express efficiently an enhanced green fluorescent protein (eGFP) in vitro. This eGFP-tagged recombinant SVA (rSVA-eGFP) exhibited a high capacity for viral replication. Its fluorescence-tracked characteristics greatly facilitated both virus neutralization test (VNT) and antiviral assay. The rSVA-eGFP-based VNT was used to detect eight porcine serum samples, out of which four were determined to be neutralization titer-positive. Subsequently, two antiviral drugs, ribavirin and apigenin, were assayed for evaluating both effects against the rSVA-eGFP in vitro. The result showed that only the ribavirin exhibited an anti-SVA activity.

Keywords: Senecavirus A; apigenin; eGFP; reverse genetics; ribavirin; virus neutralization test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Antibody Technique
  • Genes, Reporter*
  • Genetic Engineering*
  • Green Fluorescent Proteins* / genetics
  • Neutralization Tests
  • Picornaviridae / drug effects*
  • Picornaviridae / genetics*
  • Picornaviridae / immunology
  • Reverse Genetics

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins

Supplementary concepts

  • Senecavirus A