This study aimed to evaluate the resistance of 79 sugarcane genotypes (varieties and elite clones) artificially inoculated with a severe Sugarcane mosaic virus (SCMV) strain and to estimate genetic parameters associated with SCMV resistance. Symptoms were evaluated using a graded scale, and confirmation of infection was performed using the Plate Trapped Antibody-ELISA serological test in plants conducted in a greenhouse experiment and then validated under field conditions. The mean incidence of mosaic disease was low in greenhouse conditions, with higher observed incidence values when assessed in the field. Based on the field results, variance analysis was significant for genotypes and genotype × environment interaction. Tracking mosaic symptom expression over time in the different sugarcane genotypes revealed varying behavior, including recovery from disease in some genotypes. The broad-sense heritability based on the mean and individual level was 62.18% and 19.37%, respectively. Twenty-two genotypes were observed to be resistant to SCMV.