CRISPR-Cas13 Based Point-of-Care Diagnostics for Rapid Detection of Antimicrobial Resistance Genes

Elena Vasquez; Wei Huang; James Okafor

doi:10.55001/gast.v1i2.63

Abstract

Antimicrobial resistance (AMR) poses a critical global health threat, yet conventional culture-based susceptibility testing requires 48-72 hours. We developed a CRISPR-Cas13a lateral flow assay (CRISPR-POC) that detects five clinically relevant resistance genes (blaKPC, blaNDM, mecA, vanA, mcr-1) directly from clinical specimens within 45 minutes. The platform combines isothermal RPA pre-amplification with Cas13a collateral cleavage of FAM-BHQ-labeled reporters, producing visible test-line signals on lateral flow strips. Clinical validation on 312 urine and wound swab samples achieved 96.8% sensitivity and 98.2% specificity compared to whole-genome sequencing, with a limit of detection of 50 copies/reaction. The assay operates at room temperature after a single 37°C incubation, making it suitable for resource-limited settings and emergency departments.

Author Biographies

Elena Vasquez Department of Microbiology, University of Barcelona, 08028 Barcelona, Spain

Elena Vasquez is an associate professor at Department of Microbiology, University of Barcelona, 08028 Barcelona, Spain. Their research focuses on biomedical engineering, with over 39 publications in peer-reviewed journals.
Wei Huang State Key Laboratory of Diagnostic Medicine, Chongqing Medical University, Chongqing 400016, China

Wei Huang is an assistant professor at State Key Laboratory of Diagnostic Medicine, Chongqing Medical University, Chongqing 400016, China. Their research focuses on social sciences, with over 46 publications in peer-reviewed journals.
James Okafor Institute of Infectious Diseases, University of Lagos, Lagos 101017, Nigeria

James Okafor is an assistant professor at Institute of Infectious Diseases, University of Lagos, Lagos 101017, Nigeria. Their research focuses on biomedical engineering, with over 29 publications in peer-reviewed journals.