A Rapid Antimicrobial Resistance Diagnostic Platform for Staphylococcus aureus Using Recombinase Polymerase...

Area of Science:

• Microbiology and Infectious Diseases
• Molecular Diagnostics
• Antimicrobial Resistance

Background:

• Antimicrobial resistance (AMR) in Staphylococcus aureus is a significant global public health threat.
• There is a critical need for rapid and accurate diagnostic methods to detect S. aureus and its AMR genes.
• S. aureus causes common nosocomial and community-acquired infections, necessitating swift diagnosis and treatment.

Purpose of the Study:

• To develop and validate a rapid diagnostic platform for simultaneous detection of S. aureus and its clinically relevant AMR genes.
• To assess the performance of recombinase polymerase amplification (RPA) for identifying S. aureus and multiple AMR markers.
• To provide an accessible diagnostic tool for low-resource and professional-lacking settings.

Main Methods:

• Development of two RPA-based detection versions: fluorescent signal monitoring and lateral flow dipstick.
• Detection of the specific S. aureus nuc gene and eight other AMR genes representing four antibiotic families.
• Validation of sensitivity, specificity, and accuracy using clinical samples and comparison with PCR.

Main Results:

• The RPA tool achieved high sensitivity, specificity, and accuracy (all >92%) in detecting AMR for all 54 S. aureus isolates.
• RPA results demonstrated 100% consistency with conventional PCR methods.
• The platform identified S. aureus and 10 different AMR genes within 40 minutes.

Conclusions:

• A rapid, accurate, and sensitive diagnostic platform for S. aureus and its AMR has been successfully developed using RPA.
• The RPA method is a viable and effective tool for clinical microbiology laboratories to guide antibiotic therapy.
• This adaptable diagnostic platform can address the shortage of rapid detection tools for drug-resistant S. aureus infections.