<p>The invasive Asian hornet (<i>Vespa velutina nigrithorax</i>) continues its spread across Europe, posing a significant threat to biodiversity, viticulture, and apiculture. This study reports the first molecular data of the invasive yellowlegged Asian hornet (<i>Vespa velutina nigrithorax</i>) in Slovakia, confirmed through molecular analysis of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Radio telemetry successfully located the nest within inaccessible private property, highlighting the technique’s crucial role in early detection. This finding, along with the need for manual tracking techniques, public awareness campaigns, and regional monitoring programs, underscores the urgent need for proactive legal frameworks to facilitate the use of radio telemetry and ensure timely intervention to prevent further spread and mitigate the ecological and economic impacts of this invasive species in Slovakia and neighboring countries. Furthermore, this study emphasizes the importance of continued research and development of radio telemetry techniques, including improved signal range and integration with drone technology, to enhance the efficiency and effectiveness of <i>V. velutina</i> detection and control.</p>