Paper Microfluidic Platform Using Multiplexed Isothermal Amplification and CRISPR/Cas12a for Aquatic Pathogen...

Area of Science:

• Environmental microbiology
• Biosensor technology
• Point-of-care diagnostics

Background:

• Microbial contamination in aquatic systems poses a global health risk.
• Current pathogen detection methods are often expensive and require specialized labs, limiting point-of-care testing (POCT).
• There is a need for accessible, field-deployable pathogen monitoring solutions.

Purpose of the Study:

• To develop an integrated paper microfluidic platform for simultaneous detection of key pathogenic bacteria.
• To enable rapid, low-cost, and portable pathogen monitoring for environmental and food safety.
• To provide a feasible POCT solution, especially for resource-limited settings.

Main Methods:

• Integrated paper microfluidic chip housing LAMP, RAA-CRISPR, and RPA-CRISPR assays.
• Engineered HRP cascade-coupled crRNA modification system for colorimetric detection.
• Solar-powered portable hardware for incubation/imaging and a web application for analysis.

Main Results:

• Spatially multiplexed detection of seven significant pathogenic bacteria (e.g., Salmonella, E. coli).
• Achieved limit of detection (LOD) of 1 CFU/mL, dynamic range of 1-10^7 CFU/mL, and high reproducibility (CV <5%).
• Demonstrated low cost (£2.5/test), 60-min sample-to-answer time, and successful field validation (R^2 = 0.98 vs. PCR).

Conclusions:

• The developed platform offers a cost-effective, sensitive, and rapid POCT solution for pathogenic bacteria detection.
• Its portable design and field validation confirm practical feasibility for diverse aquatic environments.
• This technology holds significant promise for public health protection and epidemic surveillance, particularly in resource-limited areas.