Journal of Agronomy and Field Innovations

Background:  Ralstonia solanacearum is a pathogen responsible for bacterial wilt, a disease that affects solanaceous crops. Current methods for diagnosing R. solanacearum infection are slow, taking anywhere from 2 to 7 days for confirmation.
Objective: The goal of this study was to develop a fast, sensitive, and field-friendly RPA assay that could accurately detect R. solanacearum.
Methods: RSC0707 gene primers were designed and standardized to enable rapid RPA application (37°C for 15 minutes). Sensitivity was determined using serial dilutions (10¹-10⁸ CFU/mL). Specificity was determined by testing with 12 relevant bacterial species. The field validation was conducted using a variety of samples.
Results: The RPA assay can detect the presence of 10² CFU/mL or more of R. solanacearum, which is similar to PCR. The results of the RPA assay can be obtained within just 15 minutes with 98.3% sensitivity and 99.1% specificity in diagnosis. The accuracy of the RPA in analyzing 150 field samples was 95.2% when compared with culture results. There was no evidence of RPA cross-reactivity with other species.
Conclusion: This newly developed RPA demonstrates the ability to quickly provide accurate detection of R. solanacearum and that it possesses numerous advantages over traditional detection methods such as field-deployability.