ArticleThe Journal of veterinary medical science2026
Development of a conformation-specific nanobody-based ELISA for quantitative detection of intact Senecavirus A virions and virus-like particles.
Article in The Journal of veterinary medical science, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Replication of Senecavirus A (SVA) generates intact virions and empty capsids, some of which disassemble into pentamers. To enable precise quality control of SVA intact viral antigen, we established a double-antibody sandwich ELISA to quantify intact SVA virions and virus-like particles (VLPs). The assay employs a conformation-specific nanobody (SV1) as the capture antibody, which binds both virions and VLPs, and a His-tagged nanobody (SVH1) as the detection antibody. Using purified virions as standards, the assay demonstrated a limit of detection (LOD) of 9.7 ng/mL, with a linear range spanning from 9.7 ng/mL to 1.25 μg/mL. Critically, it showed no cross-reactivity with pentamers and other viral particles, including foot-and-mouth disease virus (FMDV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), classical swine fever virus (CSFV), and encephalomyocarditis virus (EMCV). The coefficients of variation (CV) were <10% for both intra-batch and inter-batch analyses. Validation against sucrose density gradient centrifugation (SDGC) confirmed concordant results. Applied to vaccine development, the ELISA quantified a 36% antigen loss in SVA VLPs after emulsification. In summary, this method provides a rapid, specific tool for SVA vaccine standardization.
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