Cymbidium ringspot virus (CymRSV) can pose a significant threat to both the orchid and legume industries due to its wide host range and diverse modes of transmission. Rapid, sensitive, accurate, and easy-to-use detection of CymRSV is crucial for controlling its spread. This study developed a recombinase polymerase amplification (RPA) assay for the specific detection of CymRSV under isothermal conditions. The assay operates at a constant temperature of 39°C for 30 min without the need for thermal cycling. The method demonstrated high specificity, with no cross-reactivity to other viruses affecting orchids or related families. Sensitivity testing revealed a detection limit of 10 2 copies or 0.3 fg, indicating that the designed specific primers were highly sensitive to virus detection. This RPA-based detection technique offers a reliable solution for rapid diagnostics in quarantine settings, as well as in laboratories and field environments where advanced precision instruments may not be available.
Wang et al. (Mon,) studied this question.