AUTHOR=Jin Li , Hu XiaoFeng , Tian Yuan , Fang MengYa , Dong Xue , Jiang YaXuan , Han Yao , Li Hao , Sun Yansong 

TITLE=Detection of Staphylococcus aureus virulence gene pvl based on CRISPR strip

JOURNAL=Frontiers in Immunology

VOLUME=15

YEAR=2024

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1345532

DOI=10.3389/fimmu.2024.1345532

ISSN=1664-3224

ABSTRACT=<sec><title>Introduction</title><p><italic>Staphylococcus aureus</italic> (<italic>S. aureus</italic>) 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 <italic>pvl</italic>-positive <italic>S. aureus</italic> often exhibit more severe symptoms and carry a substantially higher mortality risk. Therefore, it is crucial to promptly and accurately detect <italic>pvl</italic>-positive <italic>S. aureus</italic> before initiating protective measures and providing effective antibacterial treatment.</p></sec><sec><title>Methods</title><p>In this study, we propose a precise identification and highly sensitive detection method for <italic>pvl</italic>-positive <italic>S. aureus</italic> based on recombinase-assisted amplification and the CRISPR-ERASE strip which we previously developed.</p></sec><sec><title>Results</title><p>The results revealed that this method achieved a detection limit of 1 copy/μL for <italic>pvl</italic>-positive plasmids within 1 hour. The method successfully identified all 25 <italic>pvl</italic>-positive and 51 <italic>pvl</italic>-negative strains among the tested 76 isolated <italic>S. aureus</italic> samples, demonstrating its concordance with qPCR.</p></sec><sec><title>Discussion</title><p>These results show that the CRISPR-ERASE detection method for <italic>pvl</italic>-positive <italic>S. aureus</italic> 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 <italic>pvl</italic>.</p></sec>