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

Yuwei Pan^1,2, Zhugen Yang¹

¹Faculty of Engineering and Applied Sciences, Cranfield University, Cranfield MK43 0AL, UK.

|July 7, 2026

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This study introduces a low-cost, paper-based microfluidic platform for rapid detection of multiple pathogenic bacteria in water. The system offers a feasible point-of-care testing (POCT) solution for public health and environmental monitoring.

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.

Keywords:

CRISPR/Cas12a aquatic monitoring isothermal amplification multiplexed detection paper microfluidic platform point-of-care testing (POCT)

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Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

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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.