A new method to discriminate between tobamoviruses and their pathotypes that infect green pepper (Capsicum annuum L.) was developed using reverse transcription-polymerase chain reaction (RT-PCR). The P0 pathotype (Tobacco mosaic virus, Tomato mosaic virus, and Tobacco mild green mosaic virus) and the P1 pathotype (Paprika mild mottle virus) were distinguished by RT-PCR using primers specific to each pathotype. However, the P1,2 and P1,2,3 pathotypes of Pepper mild mottle virus (PMMoV) could not be distinguished from each other using this procedure. The P1,2 and P1,2,3 pathotypes were differentiated by RT-nested PCR, in which a DNA fragment was first produced by RT-PCR using primers containing conserved sequences of PMMoV. The product was then used as the DNA template in a second PCR using primers specific to each pathotype. An immunocapture (IC) RT-PCR method was developed based on results from this study, which facilitated detection of tobamoviruses in pepper plants, seeds, and field soils and allowed the identification of their pathotypes.