Detection of eight different tospovirus species by a monoclonal antibody against the common epitope of NSs protein

Rabbit antisera against the nucleocapsid protein (NP) have been commonly used for detection of tospoviruses and classification into serogroups or serotypes. Mouse monoclonal antibodies (MAbs) with high specificity to the NPs have also been widely used to identify tospovirus species. Recently, a serogroup-specific MAb against the NSs protein of Watermelon silver mottle virus (WSMoV) was produced by our laboratory to react with five members of WSMoV serogroup, i.e., WSMoV, Capsicum chlorosis virus (CaCV), Calla lily chlorotic spot virus (CCSV), Peanut bud necrosis virus (PBNV) and Watermelon bud necrosis virus (WBNV). The epitope recognized by the NSs MAb was determined and the comparison with the reported sequences of tospoviral NSs proteins revealed that the epitope is highly conserved at the N-terminal region of NSs proteins among members of WSMoV and Iris yellow spot virus (IYSV) serogroups, and Melon yellow spot virus (MYSV) serotype. When the NSs MAb was further used to react with the crude antigens of MYSV serotype, IYSV and Tomato yellow ring virus (TYRV) of IYSV serogroup, strong serological reactions, both in ELISA and western blotting, were observed. Thus, our results indicated that the NSs MAb is a useful and convenient tool for detection of the eight tospovirus species. It is also suggested that these eight Asian-type tospoviruses, i.e., WSMoV, CaCV, CCSV, PBNV, WBNV, MYSV, IYSV and TYRV, may share a common evolutionary ancesto