Tomato yellow leaf curl virus (TYLCV) is the most widespread virus species causing Tomato yellow leaf curl disease. Accurate methods for assessment of resistance are requisite in breeding programs. Researchers at the Volcani Center (Israel) developed a scale of differential TYLCV reactions in seven non-segregating tomato host lines, with virus responses ranging from susceptible to highly resistant. In the present work, this scale has been evaluated with inoculation and assessment methods routinely used by two different research groups, in Spain and Cuba. Different TYLCV isolates, inoculation methods, plant ages, and environmental conditions were compared. Symptom scores of the tomato lines were generally lower in the conditions assayed in both locations than those originally described. Reaction ranking order of the standard tomato lines was more similar to the original description in the assays carried out in Spain. However, response of the lines to TYLCV obtained in both locations did not correspond to scale grades for most of the conditions. A large difference between symptom scores in the susceptible and the resistant lines was observed, with the range in the resistant lines being narrow. These results indicate that the number of standard lines used could be reduced, selecting the most susceptible and the most resistant lines, and one with intermediate resistance. All the factors evaluated affected symptom development. This highlights the importance of establishing a standard inoculation method, experimental conditions, evaluation period and appropriate resistance evaluation criteria, to ensure precise evaluation of genotype responses