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Arterial hypertension (HTN) is a growing global health concern, with limited tools available for early detection. Previous studies identified the overexpression of the epithelial sodium channel (ENaC) as a potential biomarker for HTN. In this work, we optimized and clinically validated a lateral flow immunoassay (LFIA) using gold nanoparticles (AuNPs) functionalized with anti-ENaC antibodies. The test strips were prepared with 10 µL of each component and performed in a 9-point herringbone format. For validation, a double-blind study was conducted using platelet lysates from 200 individuals, classified based on real-time blood pressure measurements. ENaC expression was assessed via both LFIA and Western blotting, which served as the reference method. Receiver operating characteristic (ROC) analysis yielded an AUC of 0.7314 for LFIA and 0.6491 for the Western blot, with LFIA demonstrating higher sensitivity (76.24%) and comparable specificity (61.54%) compared to the Western blot (68.31% and 60.34%, respectively). These results support LFIA as a practical, rapid, and moderately accurate tool for screening ENaC levels and identifying individuals at risk of hypertension.