A microfluidic enrichment platform with a recombinase polymerase amplification sensor for pathogen diagnosis

Area of Science:

• Biomedical Engineering
• Microfluidics
• Pathogen Detection

Background:

• Early and sensitive detection of pathogens is crucial for patient diagnosis, treatment, and public health surveillance.
• Existing methods often struggle with low pathogen concentrations in large sample volumes.
• There is a need for advanced technologies to enhance pathogen capture and detection efficiency.

Purpose of the Study:

• To develop and validate a microfluidic platform for sensitive detection of low pathogen levels in large sample volumes.
• To integrate a Magainin 1-based enrichment system with a recombinase polymerase amplification (RPA) sensor.
• To achieve rapid, label-free, and real-time detection of pathogenic DNA.

Main Methods:

• Utilized a herringbone microfluidic chip functionalized with Magainin 1 for pathogen enrichment.
• Coupled the enrichment platform with a recombinase polymerase amplification (RPA) sensor for DNA amplification and detection.
• Employed an isothermal DNA amplification method for rapid and sensitive pathogen identification.
• Performed detection in 10 ml urine samples for Salmonella and Brucella.

Main Results:

• The Magainin 1 enrichment platform demonstrated a 20-fold increase in sensitivity for Salmonella and a 10-fold increase for Brucella in urine compared to real-time PCR without enrichment.
• The combined system detected down to 50 CFU/10 ml urine for Salmonella and 10^2 CFU/10 ml urine for Brucella within 60 minutes.
• Achieved label-free and real-time detection of pathogenic DNA.

Conclusions:

• The developed microfluidic platform significantly enhances pathogen detection sensitivity by improving capture efficiency in large sample volumes.
• This integrated system offers a promising tool for improved early diagnosis and surveillance of infectious diseases.
• The platform's rapidity and sensitivity make it suitable for clinical diagnostics and public health applications.