A quick and on-the-spot applicable method for the strain-specific detection of wheat dwarf virus by LAMP technique

Wheat dwarf virus (WDV) is a major cereal pathogen comprising two economically important strains, WDV-wheat (WDV-W) and WDV-barley (WDV-B), whose rapid and reliable differentiation is essential for effective disease management. In this study, we developed and validated novel strain-specific loop-mediated isothermal amplification (LAMP) assays for the detection of WDV directly from plant and vector samples. The optimized assays enabled clear discrimination between WDV-W and WDV-B within 30–60 min, using a simple boiling-based DNA preparation without the need for commercial extraction kits. Under laboratory conditions, both primer sets showed high specificity with no cross-reactivity. Sensitivity tests revealed that the WDV-B LAMP assay was up to 10 4 -fold more sensitive than conventional PCR, while the WDV-W assay showed approximately 10-fold higher sensitivity. Field trials demonstrated that the entire workflow, including sample preparation, amplification and result evaluation can be performed on-site within 1–2 h using portable equipment, with consistent and reliable results under realistic environmental conditions. Additionally, a practical outdoor LAMP protocol was developed and is provided as supplementary material. The proposed method offers a rapid, cost-effective and extraction-free approach for strain-specific WDV detection at the point of need, with strong potential to support field surveillance, early disease detection and timely crop protection decisions. • Strain-specific isothermal assays developed for wheat dwarf virus • High sensitivity surpassing conventional PCR methods • Rapid detection within 30–60 min using portable equipment • Outdoor LAMP protocol established for on-site diagnostics