Abstract
Background Senecavirus A (SVA), a member of the family Picornaviridae, genus Senecavirus, is a recently identified single-stranded RNA virus closely related to members of the Cardiovirus genus. SVA was originally identified as a cell culture contaminant and was not associated with disease until 2007 when it was first observed in pigs with Idiopathic Vesicular Disease (IVD). Vesicular disease is sporadically observed in swine, is not debilitating, but is significant due to its resemblance to foreign animal diseases, such as foot-and-mouth disease (FMD), whose presence would be economically devastating to the United States. IVD disrupts swine production until foreign animal diseases can be ruled out. Identification and characterization of SVA as a cause of IVD will help to quickly rule out infection by foreign animal diseases.MethodsWe have developed and characterized an indirect ELISA assay to specifically identify serum antibodies to SVA. Viral protein 1, 2 and 3 (VP1, VP2, VP3) were expressed, isolated, and purified from E. coli and used to coat plates for an indirect ELISA. Sera from pigs with and without IVD symptoms as well as a time course following animals from an infected farm, were analyzed to determine the antibody responses to VP1, VP2, and VP3.ResultsAntibody responses to VP2 were higher than VP1 and VP3 and showed high affinity binding on an avidity ELISA. ROC analysis of the SVA VP2 ELISA showed a sensitivity of 94.2% and a specificity of 89.7%. Compared to IFA, the quantitative ELISA showed an 89% agreement in negative samples and positive samples from 4–60 days after appearance of clinical signs. Immune sera positive for FMDV, encephalomyocarditis virus, and porcine epidemic diarrhea virus antibodies did not cross-react.ConclusionsA simple ELISA based on detection of antibodies to SVA VP2 will help to differentially diagnose IVD due to SVA and rule out the presence of economically devastating foreign animal diseases.
Highlights
Senecavirus A (SVA), a member of the family Picornaviridae, genus Senecavirus, is a recently identified single-stranded RNA virus closely related to members of the Cardiovirus genus
SVA was originally identified as a cell culture contaminant and was not associated with disease until 2007 when it was first observed in pigs with Idiopathic Vesicular Disease (IVD) [2, 3]
ELISA plates were coated with 500 ng/well of VP1, VP2 or VP3 protein and samples were examined for anti-SVA antibody reactivity
Summary
Senecavirus A (SVA), a member of the family Picornaviridae, genus Senecavirus, is a recently identified single-stranded RNA virus closely related to members of the Cardiovirus genus. Identification and characterization of SVA as a cause of IVD will help to quickly rule out infection by foreign animal diseases. Senecavirus A (SVA), a member of the family Picornaviridae, genus Senecavirus, is a recently identified singlestranded RNA virus closely related to members of the Cardiovirus genus [1, 2]. IVD in association with SVA has been observed recently in Canada, the United States, and Brazil, in the absence of other vesicular foreign animal diseases [3, 5, 6]. SVA infection is diagnosed by RT-PCR, a serum neutralizing assay, indirect fluorescent antibody test (IFA), or competitive enzyme-linked immunosorbent assay (cELISA) [6–9]. Numerous ELISA kits used for the detection of viral antibodies are commercially available, an indirect ELISA kit is not yet commercially available for the detection of anti-SVA antibodies in pigs
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