Rapid screening of ten tobamoviruses based on reverse-transcription recombinase-aided amplification and lateral flow dipstick.

Background

Tobamoviruses infect economically important crops in the Solanaceae and Cucurbitaceae families, causing substantial economic losses. The genus Tobamovirus comprises several highly destructive viruses, such as tobacco mosaic virus (TMV), cucumber green mottle mosaic virus (CGMMV), tomato mosaic virus (ToMV), tomato mottle mosaic virus (ToMMV), pepper mild mottle virus (PMMoV) and tomato brown rugose fruit virus (ToBRFV). Infections caused by these viruses pose serious threats to crop yield and quality, resulting in substantial economic losses. Therefore, rapid and accurate detection of tobamoviruses is crucial for controlling their spread.

Results

A broad-spectrum detection method for important tobamoviruses was developed based on reverse-transcription recombinase-aided amplification (RT-RAA) combined with lateral flow dipstick (LFD). This RT-RAA-LFD method enables the rapid determination of tobamovirus presence in samples via test strip observation following a 20-min isothermal reaction at 42 °C. The assay can detect ten virus species in the genus Tobamovirus, namely TMV, ToMV, ToMMV, ToBRFV, PMMoV, CGMMV, cucumber fruit mottle mosaic virus (CFMMV), cucumber mottle virus (CMoV), paprika mild mottle virus (PaMMV) and Rehmannia mosaic virus (ReMV). The detection limits of this method for ToBRFV, CGMMV and PMMoV were approximately 100 times lower than those of a reverse-transcription-polymerase chain reaction. This method exhibits high stability and detection efficiency, with no cross-reactivity against other common viruses infecting Solanaceae and Cucurbitaceae plants.

Conclusion

This study developed a broad-spectrum RT-RAA-LFD-based detection method for rapid screening of ten tobamoviruses. Our study provides a useful reference for the high-throughput screening of multiple pests in a rapid and simultaneous manner. © 2026 Society of Chemical Industry.