💡 Application Note: Rapid detection using RPA-CRISPR/Cas12a. This protocol validates the use of Universal Lateral Flow Strips, making it an ideal reference for Due Bio product users.
Title: Development of an RPA-based CRISPR/Cas12a assay in combination with a lateral flow strip for rapid detection of toxigenic Fusarium verticillioides in maize
Source: Food Control (Elsevier) (2024)
Abstract
Fusarium verticillioides is a major maize pathogen producing fumonisins, posing severe health risks. This study established a rapid and sensitive detection method combining Recombinase Polymerase Amplification (RPA) with CRISPR/Cas12a and lateral flow strips (LFS). The assay targets the FUM1 gene of F. verticillioides. Under optimized conditions, the assay can detect down to 10 copies/μL of genomic DNA within 40 minutes (20 min RPA + 20 min Cas12a/LFS). The method demonstrated high specificity against other fungal pathogens and was successfully applied to maize samples.
Key Method (Tiosbio Usage)
“The CRISPR/Cas12a reaction products were detected using lateral flow strips (Tiosbio, Beijing, China). The test line (T-line) captures the FAM-labeled cleaved reporter, while the control line (C-line) captures the biotin-labeled uncleaved reporter.”