Multiplex RT-PCR to detect tospoviruses
In Japan, the following tospoviruses have been reported in crops: Tomato spotted wilt tospovirus (TSWV - EPPO A2 list) on chrysanthemums, tomato and Capsicum; Watermelon silver mottle tospovirus (WSMoV – EPPO A1 list) on watermelon and New Zealand spinach (Tetragonia tetragonioides), Impatiens necrotic spot tospovirus* (INSV – EPPO A2 list) on Cineraria, Cyclamen and Eustoma grandiflorum, and Iris yellow spot tospovirus (IYSV – EPPO Alert List) on E. grandiflorum and Alstroemeria. A one-step multiplex RT-PCR has been developed in Japan to simultaneously detect and identify these five different tospovirus species. The system is composed of a universal degenerate primer and five virus species-specific primers. With this method, it was possible to detect single infections and also multiple infections in ornamental crops.
* The EPPO Secretariat had previously no data on the occurrence of INSV in Japan.
Uga H, Tsuda S (2005) A one-step reverse transcription-polymerase chain reaction system for the simultaneous detection and identification of multiple tospovirus infections. Phytopathology 95(2), 166-171.