Highly Sensitive Field Detection Technology for Anthrax Based on the CRISPR/Cas13a System

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Bacillus anthracis poses a significant bioterrorism threat.
• Rapid and sensitive detection methods are crucial for timely response.
• Existing detection methods may lack portability or speed for on-site applications.

Purpose of the Study:

• To develop a highly sensitive and specific on-site detection technology for Bacillus anthracis.
• To integrate Multiple Enzyme Isothermal Rapid Amplification (MIRA) with CRISPR/Cas13a.
• To establish a portable point-of-care testing (POCT) device for B. anthracis.

Main Methods:

• Integration of MIRA with a CRISPR/Cas13a detection system.
• Optimization of crRNA selection, MIRA primers, and reaction conditions.
• Development of a CRISPR-based point-of-care testing device with lyophilized reagents.

Main Results:

• The optimized CRISPR/Cas13a assay achieved a detection limit of 1000 copies/mL for B. anthracis.
• The developed POCT device demonstrated a detection limit of 250 copies/mL within 30 minutes.
• The assay showed high specificity against common respiratory pathogens and high accuracy in simulated clinical samples.

Conclusions:

• A highly sensitive and portable CRISPR/Cas13a-based technology for on-site B. anthracis detection was successfully established.
• The developed POCT device offers rapid, accurate, and specific detection capabilities.
• This technology holds significant potential for the surveillance of emerging infectious diseases.