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Updated: Aug 5, 2026
Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria
Published on: May 8, 2013
Rapid One-Tube RPA-CRISPR/Cas12 Detection Platform for Methicillin-Resistant Staphylococcus aureus
Yanan Li1,2, Zhonglin Shi1,3, Anzhong Hu1
1Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China.
Abstract:
Methicillin-resistant Staphylococcus aureus (MRSA) is a severe health threat causing high-level morbidity and mortality in health care environments and in community settings. Though existing diagnostic methods, including PCR and culture-based methods, are routinely used in clinical practice, they are not appropriate for rapid point-of-care testing (POCT). Recently, since the development of the CRISPR/Cas technology, new possibilities for rapid point-of-care detection have emerged. In this study, we developed a rapid, accurate, and contamination-free platform for MRSA detection by integrating recombinase polymerase amplification (RPA) with the Cas12 system into one tube. Using this approach, visual MRSA detection could be achieved in 20 min. Based on the one-tube RPA-CRISPR/Cas12a platform, the assay results are visualized by lateral flow test strips (LFS) and fluorescent-based methods, including real-time and end-point fluorescence. This platform allows specific MRSA detection with a sensitivity of 10 copies for the fluorescence method and a range of 10-100 copies for the LFS. The results of 23 samples from clinical MRSA isolates showed that the coincidence rate was 100% and 95.7% of the fluorescence method and LFS, respectively, compared to qPCR. In conclusion, the one-tube RPA-CRISPR/Cas12a platform is an effective method for MRSA detection with significant potential in future practical POCT applications.
Insights
A novel one-tube RPA-CRISPR/Cas12a platform offers rapid, contamination-free detection of Methicillin-resistant Staphylococcus aureus (MRSA). This point-of-care test achieves visual MRSA detection in just 20 minutes, showing high accuracy for clinical applications.
Area of Science:
- Molecular Biology
- Biotechnology
- Infectious Disease Diagnostics
Background:
- Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant global health threat, leading to high morbidity and mortality.
- Current diagnostic methods like PCR and culture are not suitable for rapid point-of-care testing (POCT).
- CRISPR/Cas technology presents new opportunities for developing rapid POCT methods.
Purpose of the Study:
- To develop a rapid, accurate, and contamination-free platform for MRSA detection.
- To integrate recombinase polymerase amplification (RPA) with the Cas12 system for a one-tube assay.
- To enable visual detection of MRSA within 20 minutes for potential POCT applications.
Main Methods:
- Developed a one-tube platform combining RPA and CRISPR/Cas12a systems.
- Utilized lateral flow test strips (LFS) and fluorescence-based methods for result visualization.
- Evaluated assay sensitivity and specificity using known copy numbers and clinical isolates.
Main Results:
- Achieved visual MRSA detection in 20 minutes.
- Demonstrated high sensitivity: 10 copies (fluorescence) and 10-100 copies (LFS).
- Clinical isolate testing showed 100% (fluorescence) and 95.7% (LFS) coincidence with qPCR.
Conclusions:
- The one-tube RPA-CRISPR/Cas12a platform is an effective method for MRSA detection.
- The platform offers significant potential for practical point-of-care testing applications.
- This assay provides a rapid and accurate alternative for MRSA diagnostics.
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