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

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.

SUBMITTER: Liu F

PROVIDER: S-EPMC7150990 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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

Liu Fuxiao F Huang Yilan Y Wang Qianqian Q Shan Hu H

Viruses 20200305 3

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 <i>Senecavirus</i> in the family <i>Picornaviridae</i>. 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 ...[more]

PMID: 32150804

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Project description:Senecavirus A (SVA)-associated porcine idiopathic vesicular disease (PIVD) and pseudorabies (PR) are highly contagious swine diseases that pose a significant threat to the swine industry in China. Since there is currently no effective commercial vaccine against SVA, the virus has spread widely throughout China and its pathogenicity has increased over the last decade. In this study, a recombinant strain named rPRV-XJ-ΔTK/gE/gI-VP2 was constructed by using the pseudorabies virus (PRV) variant strain XJ as the parental virus and by deleting the TK/gE/gI gene while coexpressing SVA VP2. The recombinant strain can stably proliferate and express foreign protein VP2 in BHK-21 cells while having a similar virion appearance to that of the parental strain. rPRV-XJ-ΔTK/gE/gI-VP2 is safe and effective for BALB/c mice, inducing high levels of neutralizing antibodies against both PRV and SVA, providing 100% protection from the virulent PRV strain. Histopathological examination and quantitative PCR (qPCR) assay have demonstrated that SVA can infect mice through intranasal inoculation, while the vaccination of mice with rPRV-XJ-ΔTK/gE/gI-VP2 can significantly reduce SVA virus copies and alleviate the pathological inflammatory changes in the heart and liver. The evaluation of the safety and immunogenicity indicates that rPRV-XJ-ΔTK/gE/gI-VP2 holds promise as a candidate vaccine against PRV and SVA. IMPORTANCE This study reports the construction of a recombinant PRV with SVA for the first time, and the resulting virus, rPRV-XJ-ΔTK/gE/gI-VP2, can induce high levels of neutralizing antibodies against both PRV and SVA in model mice. These findings provide valuable insights for evaluating the effectiveness of rPRV-XJ-ΔTK/gE/gI-VP2 as a vaccine for pigs. Additionally, this study reports transient SVA infection in mice, with qPCR assays showing that the copies of the SVA 3D gene peaked at 3 to 6 days postinfection and fell below the sensitivity threshold by 14 days postinfection. The copies of the gene were more regular and at a higher level in the heart, liver, spleen, and lung tissue.

Dataset Information

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

Publications

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

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