FARPA-based tube array coupled with quick DNA extraction enables ultra-fast bedside detection of antibiotic-resistant...

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Infectious Disease Research

Background:

• Conventional culture-based methods for pathogen detection are time-consuming, delaying clinical diagnosis and treatment.
• Rapid identification of pathogens and antimicrobial resistance (AMR) genes is critical for effective infectious disease management.
• Current diagnostic limitations hinder the timely application of anti-infection therapies.

Purpose of the Study:

• To develop and validate a novel, ultra-fast point-of-care testing (POCT) method for detecting antibiotic-resistant pathogens.
• To improve sample throughput for simultaneous analysis of multiple pathogens and samples.
• To provide a rapid diagnostic tool for guiding clinical treatment decisions.

Main Methods:

• A tube array strategy was developed based on the FEN1-aided recombinase polymerase amplification (FARPA) principle.
• A simplified sample-to-result workflow integrated rapid DNA extraction with FARPA, completing in 25 minutes.
• A 36-tube array format was utilized with a portable analyzer for high-throughput POCT.

Main Results:

• The method achieved ultra-fast detection of antibiotic-resistant pathogens directly from urine samples.
• High sensitivities were observed for detecting specific pathogens: E. coli (92.7%), K. pneumoniae (93.8%), E. faecium (100%), and E. faecalis (88.9%).
• Specificities for all tested pathogens reached 100%, demonstrating high accuracy.

Conclusions:

• The developed FARPA-based tube array strategy offers a rapid, low-cost, and user-friendly POCT solution for pathogen detection.
• This method significantly reduces turnaround time compared to conventional methods, enabling prompt clinical intervention.
• The technology holds substantial potential for guiding rational antibiotic use and combating the rise of antimicrobial resistance.