Andes orthohantavirus (ANDV), the primary etiological agent of hantavirus pulmonary syndrome (HPS) in South America, is uniquely capable of limited human-to-human transmission, posing a significant challenge for outbreak control. Recent events, including the 2018-2019 Epuyen outbreak and the 2026 MV Hondius incident, underscore the need for rapid, lineage-specific molecular diagnostics. In this study, we present an artificial intelligence (AI)-driven framework for the design of diagnostic primers targeting the S genomic segment of the Epuyen lineage. Using an evolutionary algorithm integrated with thermodynamic evaluation via Primer3Plus, candidate primers were optimized to maximize classification accuracy while satisfying stringent biochemical constraints. The resulting primer set enables amplification of lineage-specific regions suitable for molecular characterization and surveillance. In silico validation demonstrates that the proposed primers achieve perfect discrimination between 2026 outbreak sequences and other ANDV variants. Furthermore, in silico comparison with standard protocol-based primers reveals substantially reduced sensitivity and specificity in the latter, highlighting the limitations of static diagnostic designs when applied to evolving viral populations. Overall, this work demonstrates that AI-assisted primer design provides a robust and adaptable strategy to improve viral detection, enhance outbreak tracking, and support timely public health interventions. Integrating computational optimization into diagnostic development is essential for strengthening preparedness against emerging zoonotic threats.