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

Fuxiao Liu,Yilan Huang,Qianqian Wang,Hu Shan

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.

Highlights

Senecavirus A (SVA), previously known as Seneca Valley virus, is an emerging pathogen inChina [1,2]
SVA infection is characterized by porcine vesicular lesions on coronary bands, snout and oral cavities, clinically indistinguishable from those caused by foot-and-mouth disease virus (FMDV), swine vesicular disease virus and vesicular stomatitis virus [4,5]
To facilitate linearization of the recombinant plasmid, the Not I recognition sequence was added to the 30 end of recombinant SVA (rSVA)-enhanced green fluorescent protein (eGFP) cDNA clone

Summary

Introduction

Senecavirus A (SVA), previously known as Seneca Valley virus, is an emerging pathogen in. China [1,2] It is a non-enveloped, positive-sense and single-stranded RNA virus, and classified into the genus Senecavirus in the family Picornaviridae. The SVA has been silently circulating in pig herds of the USA since 1988, SVA-caused vesicular disease was not identified in pigs until 2007 in Canada [6]. The second case of SVA infection was identified in the USA in 2010 [7]. Afterwards, SVA emergences were reported in more countries, including China [2], Thailand [9], Vietnam [10]

Methods

Results

Conclusion