Rapid and specific detection of Enterococcus faecalis with a visualized isothermal amplification method

Area of Science:

• Microbiology
• Molecular Diagnostics
• Biotechnology

Background:

• Enterococcus faecalis poses a significant hospital-acquired infection risk.
• Current detection methods for E. faecalis are often time-consuming and require specialized personnel.
• Rapid and accurate diagnostics are crucial for controlling E. faecalis transmission.

Purpose of the Study:

• To develop a rapid, accurate, and accessible detection method for Enterococcus faecalis.
• To overcome the limitations of existing diagnostic techniques for E. faecalis.

Main Methods:

• Development of a diagnostic assay combining recombinase polymerase amplification (RPA) with lateral flow strip (LFS) technology.
• Optimization of the RPA-LFS method for speed and sensitivity at a constant temperature (37°C).
• Validation of the method using 278 clinical samples and comparison with quantitative polymerase chain reaction (qPCR).

Main Results:

• The developed RPA-LFS method provides results in approximately 35 minutes.
• Achieved a limit of detection of 10 CFU/µL for E. faecalis in both pure and complex samples.
• Demonstrated high specificity, compatibility, and detection accuracy comparable to conventional qPCR methods.

Conclusions:

• The visualized isothermal amplification method (RPA-LFS) offers a rapid and accurate approach for E. faecalis detection.
• This method has potential for on-site diagnostics, improving hospital infection control strategies.
• The RPA-LFS assay presents a viable alternative to traditional, labor-intensive diagnostic methods.