Development of Recombinase Aided Amplification (RAA)-Exo-Probe Assay for the Rapid Detection of Shiga Toxin-Producing Escherichia coli.

The article focuses on the development of recombinase aided amplification (RAA)-exo-probe assays for the rapid, sensitive, and specific detection of Shiga toxin-producing Escherichia coli (STEC) by targeting the stx1 and stx2 genes. These assays demonstrated a detection limit of 1.6 × 10³ CFU/mL (32 copies/reaction), comparable to quantitative PCR (qPCR), but with a shorter runtime of about 20 minutes and without the need for expensive equipment, making them suitable for field or resource-limited settings. Validation using spiked and real meat samples (beef, pork, and mutton) showed successful detection of STEC, particularly after an overnight enrichment step, supporting their applicability for routine food safety monitoring. The study highlights the potential of RAA-exo-probe assays as practical tools for timely STEC surveillance to prevent foodborne outbreaks.