Microfluidic and Paper-Based Recombinase Polymerase Amplification Systems for Decentralized Diagnostics and...

Area of Science:

• Molecular Diagnostics
• Biotechnology
• Point-of-Care Testing

Background:

• Recombinase polymerase amplification (RPA) is a rapid, isothermal nucleic acid amplification method suitable for decentralized diagnostics.
• Microfluidic and paper-based platforms enable integrated sample handling, amplification, and readout in portable formats.
• RPA's mild temperature requirements and compatibility with microfluidics make it ideal for field-deployable molecular diagnostics.

Purpose of the Study:

• To review recent advancements in microfluidic and paper-based RPA systems.
• To highlight applications across biomedical diagnostics, food safety, environmental monitoring, and One Health.
• To discuss emerging trends in intelligent diagnostic formats and remaining challenges.

Main Methods:

• Review of integrated device architectures: centrifugal chips, capillary-driven platforms, microfluidic paper-based analysis devices (μPADs), electrochemical biosensors, CRISPR-assisted assays, digital microfluidics, and sample-to-answer cartridges.
• Examination of applications in various fields, including respiratory viruses, infectious diseases, cancer biomarkers, foodborne pathogens, and environmental surveillance.
• Analysis of emerging technologies like smartphone imaging, AI interpretation, and cloud connectivity.

Main Results:

• RPA systems are increasingly integrated into microfluidic and paper-based devices for diverse applications.
• Biomedical applications include detection of viruses, bacteria, parasites, and cancer biomarkers.
• Food safety and environmental monitoring applications cover pathogen detection, antimicrobial resistance, and airborne pathogen screening.
• Intelligent features such as AI and smartphone integration are enhancing diagnostic capabilities.
• Challenges remain in sample preparation, reagent stability, quantification, and regulatory validation.

Conclusions:

• Microfluidic and paper-based RPA platforms are advancing molecular diagnostics for field applications.
• The integration of advanced technologies like CRISPR and AI is improving diagnostic performance and accessibility.
• Addressing challenges in sample preparation, reagent stability, and regulatory pathways is crucial for widespread adoption.