Synthetic p72 Peptide-Based Indirect- Enzyme-Linked Immunosorbent Assay for Diagnosis of African Swine Fever Virus Infection

In the present study, we explored the potential diagnostic application of B cell linear peptides derived from the p72 protein of African Swine Fever Virus (ASFV) using a two-step bioinformatics approach for developing an indirect enzyme-linked immunosorbent assay (ELISA) for ASF detection. Through computational analysis, eight linear B cell epitopes with significant conservation across various ASFV genotypes were identified. These peptides were chemically synthesized and evaluated for their immunoreactivity using specific rabbit hyperimmune serum against the ASFV p72 protein. The synthesized peptides displayed notable reactivity in dot-ELISA and subsequently in indirect-ELISA. Validation of the indirect ELISA was conducted using samples collected during ASFV outbreaks in the northern states of India from 2022 to 2023, and further corroborated using a commercial kit. The P1 peptide-based indirect ELISA demonstrated a sensitivity of 100% and a specificity of 86%. Additionally, peptides P3, P4, P5, P7, and P8 exhibited a specificity of 100%, while peptides P2, P3, P4, P6, and P7 displayed a sensitivity greater than 70% in the indirect ELISA. These results underscore the diagnostic potential of p72 protein-specific B cell epitopes for the detection of ASFV infection in field settings. The developed assay holds promise for ASFV infection detection, as well as for seromonitoring and serosurveillance applications.