Sensitive and rapid detection of pathogenic bacteria from urine samples using multiplex recombinase polymerase...

Area of Science:

• Clinical Microbiology
• Biotechnology
• Medical Diagnostics

Background:

• Bacterial infections like UTIs require prompt diagnosis for effective treatment.
• Current culture-based methods for bacterial identification are slow (3-7 days), delaying treatment.
• Rapid and sensitive bacterial detection is crucial for managing severe diseases.

Purpose of the Study:

• To develop a rapid, sensitive, and multiplex method for detecting major pathogenic bacteria in urine samples.
• To overcome the limitations of time-consuming culture-based diagnostics for UTIs.
• To accelerate the diagnosis of urinary tract infections (UTIs) and bacteriuria.

Main Methods:

• Enrichment of bacteria from urine samples using a portable filter-based pipette.
• Development of a centrifugal chip integrating DNA extraction, multiplex recombinase polymerase amplification (RPA), and fluorescent detection.
• Detection of five key UTI pathogens: Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhimurium.

Main Results:

• Successful detection of E. coli, P. mirabilis, P. aeruginosa, and S. aureus within 40 minutes at 100 CFU mL-1.
• Detection of S. typhimurium within 40 minutes at 1000 CFU mL-1.
• Elimination of the need for bacterial cultivation, significantly reducing diagnosis time.

Conclusions:

• The developed chip-based method offers a promising tool for rapid and accurate bacterial identification in clinical urine samples.
• This technology can significantly improve the diagnosis and management of UTIs and bacteriuria.
• The method enables sensitive, accurate, and multiplex detection of common UTI pathogens.