A novel nanobody-based competitive ELISA using Nb84-HRP demonstrated 99% concordance (kappa = 0.97) with Western blot for detecting Senecavirus A antibodies in swine serum.
A novel nanobody-based competitive ELISA provides a rapid and reliable method for detecting Senecavirus A antibodies in swine, outperforming a commercial ELISA kit.
Effect estimate: Kappa 0.97
Senecavirus A (SVA) causes considerable economic losses in the swine industry, particularly among piglets. However, effective serological diagnostic assays for SVA are still lacking. A testing platform based on nanobody-horseradish peroxidase (Nb-HRP) fusion protein-based competitive ELISA (cELISA) for the detection of antibodies against various animal viruses is available. In this study, a rapid and reliable cELISA was established using this platform for specific detection of antibodies against SVA. Two anti-SVA nanobodies were selected from an immunized Bactrian camel using phage display technology. The Nb84-HRP fusion protein was expressed and used as a probe during the process. The assay exhibited a cut-off value of 19.4% and showed no cross-reactivity with antibodies against other swine viruses, demonstrating high specificity. The intra- and inter-assay coefficients of variation ranged from 1.44% to 3.27% and 1.36% to 8.71%, respectively. Evaluation using clinical swine serum samples showed a higher coincidence rate between the cELISA and Western blot (99%, kappa value = 0.97) than that between the cELISA and a commercial ELISA kit (84%, kappa value = 0.35). These results indicate that the cELISA developed in this study is a rapid, cost-effective, and reliable method for serological detection and surveillance of SVA infection.
Nie et al. (Sun,) conducted a other in Senecavirus A (SVA) infection (n=430). Nanobody-based competitive ELISA (Nb84-HRP) vs. Western blot and commercial ELISA kit was evaluated on Concordance rate with Western blot (Kappa 0.97). A novel nanobody-based competitive ELISA using Nb84-HRP demonstrated 99% concordance (kappa = 0.97) with Western blot for detecting Senecavirus A antibodies in swine serum.