Clustered\nRegularly Interspaced Short Palindromic\nRepeats-Mediated Surface-Enhanced Raman Scattering Assay for Multidrug-Resistant\nBacteria

Abstract

Antimicrobial resistance and multidrug\nresistance are slower-moving\npandemics than the fast-spreading coronavirus disease 2019; however,\nthey have potential to cause a much greater threat to global health.\nHere, we report a clustered regularly interspaced short palindromic\nrepeats (CRISPR)-mediated surface-enhanced Raman scattering (SERS)\nassay for multidrug-resistant (MDR) bacteria. This assay was developed\n <i>via</i> a synergistic combination of the specific gene-recognition\nability of the CRISPR system, superb sensitivity of SERS, and simple\nseparation property of magnetic nanoparticles. This assay detects\nthree multidrug-resistant (MDR) bacteria, species <i>Staphylococcus\naureus</i>, <i>Acinetobacter baumannii</i>, and <i>Klebsiella pneumoniae</i>, without purification or gene amplification\nsteps. Furthermore, MDR <i>A. baumannii</i>-infected mice\nwere successfully diagnosed using the assay. Finally, we demonstrate\nthe on-site capture and detection of MDR bacteria through a combination\nof the three-dimensional nanopillar array swab and CRISPR-mediated\nSERS assay. This method may prove effective for the accurate diagnosis\nof MDR bacterial pathogens, thus preventing severe infection by ensuring\nappropriate antibiotic treatment.