SummaryTomato spotted wilt virus (TSWV) is an important Tospovirus causing significant crop losses in tomato throughout the World. Resistance to TSWV is conferred by the Sw-5 gene. Screening for TSWV resistance under field conditions, and the development of resistant tomato genotypes based on phenotype is not only time-consuming, but also inconclusive because of variable inoculum pressure in the field. The development of molecular markers for marker-assisted selection (MAS) would help to avoid this problem and facilitate the screening of a large number of independent breeding lines. However, for MAS to be successful, the molecular markers must be reliable, easy to use, and highly reproducible. We evaluated four potential PCR-based markers, developed based on Sw-5 locus-specific sequences. All four PCR-based markers were used successfully to identify resistant and susceptible tomato genotypes, consistent with the phenotypic data. Three (NCSw-003, NCSw-007, and NCSw-011) of the four molecular markers were co-dominant, whereas one (NCSw-012) was dominant. Two of the four molecular markers (NCSw-003 and NCSw-012) were sequence-characterised amplified region (SCAR) markers. The other two (NCSw-007 and NCSw-011) were cleaved amplified polymorphic sequence (CAPS) markers. All four markers were 19 kbp from CT 220, a previously reported molecular marker. All four DNA markers were novel and may be useful to tomato breeders wishing to screen for TSWV resistance in segregating populations.