Peptide-driven lateral flow platform for detection of classical swine fever viral antigen

Classical Swine Fever (CSF) is a highly contagious viral disease that causes substantial economic losses in the swine industry. Timely detection of CSFV infection is essential for effective disease control; however, current antibody-based diagnostic methods are limited due to the delayed host immune response. In this study, we developed a novel peptide-based lateral flow assay (LFA) targeting the E2 antigen of the Classical Swine Fever Virus (CSFV) to enable rapid and field-deployable detection. A synthetic peptide derived from the immunodominant region of the CSFV E2 protein was identified through bioinformatics analysis, synthesized, and conjugated to Keyhole Limpet Hemocyanin (KLH) to enhance its immunogenicity. Polyclonal antibodies were generated in rabbits, purified, and validated via ELISA to confirm specificity to CSFV, showing no cross-reactivity with other swine viruses such as ASFV, PRRSV, PCV2, and PPV. Silver nanoparticles (AgNPs) were biosynthesized using Mangifera indica leaf extract and characterized by UV–vis spectroscopy, FTIR, and dynamic light scattering (DLS). These nanoparticles were then conjugated to the purified anti-E2 antibodies and incorporated into LFA strips. The resulting assay exhibited high sensitivity, capable of detecting CSFV antigen concentrations as low as 20 ng/ml, with excellent specificity and no cross-reactivity. This peptide-based LFA provides a rapid, reliable, and cost-effective diagnostic tool for on-site CSF surveillance and outbreak management.