Universal CRISPR‐Cas12a and Toehold RNA Cascade Reaction on Paper Substrate for Visual Salmonella Genome Detection

• DOI: 10.1002/adhm.202400508
• Publication Date: 2024-04-29T13:03:50Z
• Authors: Mahera J. Kachwala; Farishta Hamdard; Damla Cicek; Hilal Dagci; Christopher W. Smith; Nabeel Kalla; Mehmet V. Yigit
• Keywords: Recombinase Polymerase Amplification; T7 RNA polymerase

Salmonella, the most prevalent food-borne pathogen, poses significant medical and economic threats. Swift accurate on-site identification serotyping of Salmonella is crucial to curb its spread contamination. Here, a synthetic biology cascade reaction presented on paper substrate using CRISPR-Cas12a recombinase polymerase amplification (RPA), enabling programming standard toehold RNA switch for genome choice. This approach employs just one design differentiate between two different serotypes, i.e., S. Typhimurium Enteritidis, without need reengineering switch. The sensor exhibits high sensitivity, capable visually detecting as few 100 copies whole from model pathogen substrate. Furthermore, this robust assay successfully applied detect genomes in contaminated milk lettuce samples, demonstrating potential real sample analysis. Due versatility practical features, organisms can be detected by merely changing single element universal cell-free reaction.