Abstract BACKGROUND Broad bean wilt virus 2 (BBWV2) is a major viral pathogen causing significant economic losses in vegetable production. Existing detection methods often lack the speed, sensitivity, or simplicity required for effective on‐site field diagnosis. This study aimed to identify a novel BBWV2 isolate and develop a rapid, equipment‐free visual detection method suitable for field applications. RESULTS A novel BBWV2 isolate (BBWV2‐GZCa) was identified and characterized from pepper in Guizhou Province, China. We developed a one‐step visual detection assay by integrating reverse transcription‐recombinase polymerase amplification (RT‐RPA) with a CRISPR/Cas12a system, enabling readout via fluorescence or lateral flow dipstick (LFD). The assay demonstrated a detection limit of 7.5 copies/μL, which is 10 5 times more sensitive than conventional RT‐PCR. It showed no cross‐reactivity with other common pepper viruses and achieved 100% accuracy when validated using 20 field‐collected samples. CONCLUSION The entire detection process can be completed within one hour without specialized equipment, requiring only visual interpretation. This RT‐RPA‐CRISPR/Cas12a‐LFD method provides a rapid, highly sensitive, specific, and user‐friendly platform for on‐site detection of BBWV2, offering a practical tool for early diagnosis and disease management in agricultural settings. © 2026 Society of Chemical Industry.
Han et al. (Wed,) studied this question.