Melon yellow spot virus (MYSV), a tentative member of the genus Tospovirus, is considered a distinct serotype due to the lack of a serological relationship with other tospoviruses in its nucleocapsid protein (NP). Recently, a virus isolate collected from diseased watermelon in central Taiwan (MYSV-TW) was found to react with a rabbit antiserum (RAs) prepared against the NP of Watermelon silver mottle virus (WSMoV), and a monoclonal antibody (MAb) prepared against the common epitope of the NSs proteins of WSMoV-serogroup tospoviruses, but not with the WSMoV NP-specific MAb, in both enzyme-linked immunosorbent assay (ELISA) and western blotting. In this investigation, both RAs and MAb against MYSV-TW NP were produced. Results of serological tests revealed that the RAs to MYSV-TW NP reacted with the homologous antigen and the crude antigens of members of the WSMoV serogroup, including members of the formal species WSMoV and Peanut bud necrosis virus, and members of three tentative species, Watermelon bud necrosis virus, Capsicum chlorosis virus and Calla lily chlorotic spot virus. The MAb to MYSV-TW NP reacted only with the homologous antigen and the other geographic isolates of MYSV from Japan (JP) and Thailand (TH). Our results of reciprocal tests indicate that the NP and the NSs protein of MYSV are serologically related to those of WSMoV-serogroup tospoviruses. Furthermore, we show that both the MYSV NP MAb and the WSMoV NP MAb are reliable tools for identification of MYSV and WSMoV from single or mixed infection in field surveys, as verified using species-specific primers in reverse transcription-polymerase chain reaction.