Rapid and Sensitive Detection of Toxigenic Fusarium asiaticum Integrating Recombinase Polymerase Amplification, CRISPR/Cas12a, and Lateral Flow Techniques.

head blight (FHB) is a global cereal disease caused by a complex of species. Both and are the causal agents of FHB in China. is the predominant species in the Middle-Lower Reaches of the Yangtze River (MLRYR) and southwest China. Therefore, detecting in a timely manner is crucial for controlling the disease and preventing mycotoxins from entering the food chain. Here, we combined rapid genomic DNA extraction, recombinase polymerase amplification, Cas12a cleavage, and lateral flow detection techniques to develop a method for the rapid detection of . The reaction conditions were optimized to provide a rapid, sensitive, and cost-effective method for detection. The optimized method demonstrated exceptional specificity in detecting while not detecting any of the 14 other strains and 3 non- species. Additionally, it could detect DNA at concentrations as low as 20 ag/μL, allowing for the diagnosis of infection in maize and wheat kernels even after 3 days of inoculation. The developed assay will provide an efficient and robust detection platform to accelerate plant pathogen detection.