A Method for Detecting Five Carbapenemases in Bacteria Based on CRISPR-Cas12a Multiple RPA Rapid Detection Technology

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Carbapenem antibiotics are crucial for treating infections caused by multi-drug resistant bacteria.
• The emergence and spread of carbapenemase-producing bacteria pose a significant global health threat.
• Existing detection methods are often time-consuming and resource-intensive.

Purpose of the Study:

• To develop a rapid, sensitive, and specific detection technology for carbapenemase-producing bacteria.
• To provide a cost-effective solution for widespread screening in various settings.

Main Methods:

• Utilized CRISPR-Cas12a gene editing combined with multiple Recombinase polymerase amplification (RPA).
• Developed a method for direct isolation of carbapenemase genes from samples.
• Designed a simplified experimental protocol requiring only a water bath and no lid, minimizing aerosol contamination.

Main Results:

• Achieved a rapid detection time of 30 minutes.
• Demonstrated high detection sensitivity of 1.5 CFU/mL.
• Confirmed 100% specificity in identifying carbapenemase-producing bacteria.

Conclusions:

• The developed CRISPR-Cas12a based method is a rapid and effective tool for detecting carbapenemase-producing bacteria.
• Suitable for diverse settings, including low-resource environments and large-scale crowd screening.
• Has significant implications for hospital infection control, prevention, and clinical diagnosis.