Advanced One-Pot RPA-CRISPR/Cas12a Reaction with Glycerol and Betaine for High-Sensitivity Diagnosis of <i>mecA</i>-Carrying Strains in Clinical Samples.

ABSTRACT: The mecA gene confers methicillin resistance in both MRSA and MR-CoNS by encoding the PBP2a protein and poses a significant public health threat due to its resistance to beta-lactam antibiotics. Rapid and accurate detection of mecA is critical for timely treatment, reducing morbidity, and preventing its spread in healthcare settings. In this study, we developed an advanced one-pot recombinase polymerase amplification (RPA)-CRISPR/Cas12a system, enhanced with glycerol and betaine, for ultrasensitive detection of the mecA gene. Glycerol's viscosity effect prevents premature interaction between Cas12a and early amplification products, while betaine enhances nucleic acid amplification. The assay demonstrated superior sensitivity, detecting as low as 5 copies/μL of mecA<

SUBMITTER: Wang J 

PROVIDER: S-EPMC11822479 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature