Potato (Solanum tuberosum L.) is among the most economically important crops worldwide, yet its productivity acquires severe constraints from viral infections, especially the Potato virus X (PVX). This study aimed to develop an optimized double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for the rapid and reliable diagnosis of PVX in seed and field materials. Virus propagation in Datura stramonium and D. tatula enabled the preparation of purified antigen and polyclonal antiserum in rabbits. Optimization trials established the most effective dilutions for primary (1:2000) and secondary (1:5000) antibodies, achieving approximately 96% sensitivity and 94%–95% specificity. The improved ELISA demonstrated a strong diagnostic performance, ensuring early detection of PVX in potato cultivars grown under Uzbekistan conditions. The application of the test revealed high infection rates (80%–100%) in certain cultivars, whereas others, including Folva, Piskom, and Aureta, showed no infection, indicating their suitability for virus-free breeding. The developed DAS-ELISA provides a cost-effective, rapid, and reproducible diagnostic platform suitable for seed certification, monitoring, and phytosanitary programs. Its integration with RT-PCR and eco-safe vector management can further enhance potato yield stability and strengthen regional plant health protection systems.
Potato (S. tuberosum L.), Potato virus X, ELISA, antigen, isotonic solution, polyclonal antiserum, conjugate
Potato (S. tuberosum L.) yields primarily rely on virus-free seeds and recommended crop husbandry. The optimized DAS-ELISA (1:2000/1:5000) achieved ~96% sensitivity and ~94%–95% specificity, enabling PVX detection for seed certification, monitoring, and virus-free programs. Pairing ELISA with RT-PCR and vector control strengthens the phytosanitary protection and stability.