DNA aptamers targeting glycoprotein D enable specific detection of pseudorabies virus (PRV)

Pseudorabies virus (PRV, SuidAlphaherpesvirus 1) causes substantial economic losses in swine production. Here, we report the development of DNA aptamers targeting the PRV glycoprotein D (gD) through an optimized SELEX protocol. After 15 selection cycles, Apt-gD-2 demonstrated nanomolar affinity (Kd = 6.107 ± 0.476 nM) and high specificity for gD, as validated by an enzyme-linked aptamer-sorbent assay (ELASA) and fluorescence microscopy. Molecular docking revealed hydrogen bonding as the key interaction mechanism. The developed ic-ELASA achieved 83.3% concordance with qPCR in clinical samples, supporting its utility for on-farm PRV surveillance. These findings highlight the potential of aptamer-based diagnostic methods for rapid, sensitive, and onsite detection of PRV, offering a promising tool for disease control in the swine industry.