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Published on: February 14, 2022
Clustered Regularly Interspaced Short Palindromic Repeats-Mediated Surface-Enhanced Raman Scattering Assay for
Hongki Kim, Soohyun Lee, Hwi Won Seo
1Department of Chemical and Biomolecular Engineering (BK 21+ Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
Abstract:
Antimicrobial resistance and multidrug resistance are slower-moving pandemics than the fast-spreading coronavirus disease 2019; however, they have potential to cause a much greater threat to global health. Here, we report a clustered regularly interspaced short palindromic repeats (CRISPR)-mediated surface-enhanced Raman scattering (SERS) assay for multidrug-resistant (MDR) bacteria. This assay was developed via a synergistic combination of the specific gene-recognition ability of the CRISPR system, superb sensitivity of SERS, and simple separation property of magnetic nanoparticles. This assay detects three multidrug-resistant (MDR) bacteria, species Staphylococcus aureus, Acinetobacter baumannii, and Klebsiella pneumoniae, without purification or gene amplification steps. Furthermore, MDR A. baumannii-infected mice were successfully diagnosed using the assay. Finally, we demonstrate the on-site capture and detection of MDR bacteria through a combination of the three-dimensional nanopillar array swab and CRISPR-mediated SERS assay. This method may prove effective for the accurate diagnosis of MDR bacterial pathogens, thus preventing severe infection by ensuring appropriate antibiotic treatment.
Insights
A novel CRISPR-mediated assay rapidly detects multidrug-resistant bacteria using SERS and magnetic nanoparticles. This technology aids in diagnosing infections and guiding antibiotic treatment, crucial for combating antimicrobial resistance.
Area of Science:
- Biotechnology
- Microbiology
- Nanotechnology
Background:
- Antimicrobial resistance (AMR) and multidrug resistance (MDR) pose a significant global health threat.
- Current diagnostic methods for MDR bacteria can be slow and complex.
- Rapid and accurate detection is crucial for effective treatment and infection control.
Purpose of the Study:
- To develop a sensitive and rapid assay for detecting multidrug-resistant bacteria.
- To integrate CRISPR gene-editing, SERS, and magnetic nanoparticles for enhanced diagnostics.
- To validate the assay in detecting clinically relevant MDR bacterial species and in a mouse infection model.
Main Methods:
- Development of a clustered regularly interspaced short palindromic repeats (CRISPR)-mediated assay.
- Utilized surface-enhanced Raman scattering (SERS) for high sensitivity detection.
- Incorporated magnetic nanoparticles for efficient sample separation.
- Tested detection of *Staphylococcus aureus*, *Acinetobacter baumannii*, and *Klebsiella pneumoniae*.
- Demonstrated *in vivo* diagnosis in MDR *A. baumannii*-infected mice.
- Showcased on-site detection using a 3D nanopillar array swab.
Main Results:
- The assay successfully detected three key MDR bacterial species without prior purification or gene amplification.
- Successful diagnosis of MDR *A. baumannii* infection in a mouse model was achieved.
- On-site capture and detection of MDR bacteria were demonstrated, highlighting practical applicability.
Conclusions:
- The CRISPR-mediated SERS assay offers a sensitive, rapid, and specific method for MDR bacterial detection.
- This technology has the potential to significantly improve the diagnosis of bacterial infections.
- Accurate and timely diagnosis can lead to appropriate antibiotic treatment, combating the spread of antimicrobial resistance.
