Comparative analysis of VP1 epitopic variation among different isolates of foot-and-mouth disease virus type-O during an outbreak in the Punjab province of Pakistan.

Foot-and-mouth disease virus (FMDV) is a highly transmissible pathogen causing severe economic losses in the global livestock sector. Frequent outbreaks of FMDV type-O in Pakistan highlight the need for continuous genomic and antigenic surveillance to track its evolution. This study aimed to isolate and molecularly characterize FMDV type-O from an outbreak in the Punjab province of Pakistan using in vitro cell culture techniques. Samples were processed for viral isolation on a susceptible cell line, followed by RNA extraction. The VP1 gene, pivotal for antigenicity and immunogenicity, was amplified using a one-step polymerase chain reaction protocol. Purified amplicons underwent sequencing, and the nucleotide sequences were translated into amino acid sequences for further analysis. Protein three-dimensional modeling and in silico comparison were performed against the vaccinal seed strain PanAsia-2. The VP1 sequence analysis revealed notable genetic variability among the isolates, indicating adaptive evolution. Structural and antigenic modeling uncovered key differences between the field isolates and vaccinal strain, suggesting potential antigenic drift, which could undermine vaccine performance. The study underscores the dynamic evolution of FMDV type-O in Pakistan and the critical importance of ongoing genomic monitoring to refine vaccine strategies and enhance outbreak control.