TSWV IgG-BSA conjugate was diluted in 20 mM sodium phosphate buffer, containing 1% sucrose to a final concentration of 2 mg/ml and applied as a 0.5 mm thick, 26 cm line, centrally at 1.25 cm from the top and bottom ends on one side of a 2.5 × 26 cm Nitrocellulose membrane was cut into sections .Test line was coated with TSWV-IgG conjugate, which was applied to each membrane in 150 μg/ml TSWV using TLC conjugate sampler.