Development and validation of a competitive ELISA as a screening test for Senecavirus A

An article published in the Journal of Veterinary Diagnostic Investigation (JVDI) presents a competitive Enzyme-Linked ImmunoSorbent Assay (cELISA) and a virus neutralization test (VNT), both validated for the screening of Senecavirus A in a research setting, by the National Centre for Foreign Animal disease (NCFAD). The diagnostic specificity and sensitivity were 98.2% and 96.9% for the cELISA, and 99.6% (99.0–99.9%) and 98.2% (95.8–99.4%) for the VNT, respectively.

In Canada and the USA alike, Senecavirus A is a challenge for producers and veterinarians because of its clinical similarity to Food and Mouth Disease (FMD). Indeed, Senecavirus A, is a causative agent of swine vesicular disease with lesions developing on the snout, around the mouth and on the coronary band of the feet. Therefore, being able to differentiate Senecavirus A infections from FMD rapidly is of utmost importance to be able to take the appropriate measures.

The University of Minnesota, Veterinary Diagnostic Laboratory has developed an ImmunoFluorescence Assay (IFA) to detect antibodies against Senecavirus A. This test was used as a reference for the validation of the cELISA and VNT established by Drs. Goolia, Yang, Babiuk, and Nfon from NCFAD in collaboration with Drs. Vannucci and Patnayak from the UMN-VDL.

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Abstract: Senecavirus A (SVA; family Picornaviridae) is a nonenveloped, single-stranded RNA virus associated with idiopathic vesicular disease (IVD) in swine. SVA was detected in pigs with IVD in Brazil, United States, Canada, and China in 2015, triggering the need to develop and/or validate serologic assays for SVA. Our objective was to fully validate a previously developed competitive enzyme-linked immunosorbent assay (cELISA) as a screening test for antibodies to SVA. Additional objectives included the development and validation of a virus neutralization test (VNT) as a confirmatory test for SVA antibody detection, and the comparison of the cELISA, VNT, and an existing immunofluorescent antibody test (IFAT) for the detection of SVA antibodies in serial bleeds from SVA outbreaks. The diagnostic specificity and sensitivity were 98.2% (97.2–98.9%) and 96.9% (94.5–98.4%) for the cELISA, and 99.6% (99.0–99.9%) and 98.2% (95.8–99.4%) for the VNT, respectively. There was strong agreement among cELISA, VNT, and IFAT when compared based on kappa coefficient. Based on these performance characteristics, these tests are considered suitable for serologic detection of SVA in pigs.

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Investigation of Senecavirus A pathogenesis in finishing pigs

Dr. Fabio Vannucci, a University of Minnesota swine pathologist and his graduate student Dr. Talita Resende collaborated with a team from South Dakota State University to study the pathogenesis of Senecavirus A in finishing pigs. The results of their experiments were published online a few weeks ago in the Journal of General Virology and the printed version should be following shortly.

The importance of Senecavirus A in swine production resides in a striking resemblance in clinical signs with Food and Mouth Disease. Indeed, Senecavirus A causes vesicular lesions around the mouth and on the feet of pigs.

The collaborative work showed that Senecavirus A viremia occurred between 3 to 10 days post-inoculation (dpi), and that the neutralizing antibody response started 5 dpi. Clinical signs first observed 4dpi, lasted up to 10 days.
This study advances our understanding of Senecavirus A pathogenesis to hopefully be able to better manage it in the future.

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Abstract: Senecavirus A (SVA) is an emerging picornavirus that has been recently associated with vesicular disease and neonatal mortality in swine. Many aspects of SVA infection biology and pathogenesis, however, remain unknown. Here the pathogenesis of SVA was investigated in finishing pigs. Animals were inoculated via the oronasal route with a contemporary SVA strain SD15-26 and monitored for clinical signs and lesions associated with SVA infection. Viremia was assessed in serum and virus shedding monitored in oral and nasal secretions and feces by real-time reverse transcriptase PCR (RT-qPCR) and/or virus isolation. Additionally, viral load and tissue distribution were assessed during acute infection and following convalescence from disease. Clinical signs characterized by lethargy and lameness were first observed on day 4 pi and persisted for ~2-10 days. Vesicular lesions were observed on the snout and feet, affecting the coronary bands, dewclaws, interdigital space and heel/sole of SVA-infected animals. A short-term viremia was detected between days 3-10 post-inoculation (pi), whereas virus shedding was detected between days 1-28 pi in oral and nasal secretions and feces. Notably, RT-qPCR and in situ hybridization (ISH) performed on tissues collected on day 38 pi revealed the presence of SVA RNA in the tonsil of all SVA infected animals. Serological responses to SVA were characterized by early neutralizing antibody responses (5 days pi), which coincided with a progressive decrease in the levels of viremia, virus shedding and viral load in tissues. This study provides significant insights on the pathogenesis and infectious dynamics of SVA in swine.

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