Detection of virulence gene based on CRISPR strip.

INTRODUCTION

() is a prominent pathogen responsible for both hospital-acquired and community-acquired infections. Among its arsenal of virulence factors, Panton-Valentine Leucocidin (PVL) is closely associated with severe diseases such as profound skin infections and necrotizing pneumonia. Patients infected with -positive often exhibit more severe symptoms and carry a substantially higher mortality risk. Therefore, it is crucial to promptly and accurately detect -positive before initiating protective measures and providing effective antibacterial treatment.

METHODS

In this study, we propose a precise identification and highly sensitive detection method for -positive based on recombinase-assisted amplification and the CRISPR-ERASE strip which we previously developed.

RESULTS

The results revealed that this method achieved a detection limit of 1 copy/μL for -positive plasmids within 1 hour. The method successfully identified all 25 -positive and 51 -negative strains among the tested 76 isolated samples, demonstrating its concordance with qPCR.

DISCUSSION

These results show that the CRISPR-ERASE detection method for -positive has the advantages of high sensitivity and specificity, this method combines the characteristics of recombinase-assisted amplification at room temperature and the advantages of ERASE test strip visualization, which can greatly reduce the dependence on professional laboratories. It is more suitable for on-site detection than PCR and qPCR, thereby providing important value for rapid on-site detection of .