A microfluidic platform for multiplexed, equipment-free nucleic acid detection and point-of-need diagnostics

Background

Nucleic acid detection is a cornerstone of modern diagnostics, enabling the identification of pathogens, genetic mutations, and other biomarkers critical for disease surveillance and management. Traditionally, methods such as polymerase chain reaction (PCR) have set the standard for sensitivity and specificity, but these techniques require sophisticated laboratory infrastructure, expensive equipment, and highly trained personnel. In resource-limited or field settings, such requirements are prohibitive, creating a pressing need for portable, easy-to-use, and highly multiplexed diagnostic platforms. The growing threat of emerging infectious diseases, antimicrobial resistance, and the need for rapid outbreak response further underscore the importance of developing deployable solutions that can deliver accurate results at the point of care.

Current approaches to multiplexed nucleic acid detection face significant limitations. Rapid antigen tests, while simple and equipment-free, often lack the sensitivity required for early detection or for distinguishing closely related pathogens. Isothermal amplification methods, such as Recombinase Polymerase Amplification (RPA), reduce the need for thermal cycling but struggle with specificity, particularly at the single-nucleotide level. CRISPR-based diagnostics offer high specificity and sensitivity, but existing multiplexed implementations typically depend on complex droplet microfluidics or automated liquid handling systems, which are impractical outside of laboratory environments. Furthermore, many microfluidic devices suffer from issues such as non-uniform reagent distribution, high dead volume, and cross-contamination between reaction chambers, all of which compromise assay reliability and usability in the field. These challenges highlight the need for robust, user-friendly platforms that can deliver multiplexed, high-sensitivity nucleic acid detection without reliance on specialized equipment or infrastructure.

Technology

This technology is a portable, equipment-free microfluidic platform designed for highly multiplexed nucleic acid detection at the point of need. It features a hybrid manifold architecture that distributes a sample across up to 100 reaction chambers using a combination of parallel and serial flow geometries, all actuated via manual syringe injection. Surface tension-based valves and geometric antechambers ensure uniform, sequential filling of chambers while minimizing dead volume and preventing cross-contamination. Each chamber is pre-loaded with lyophilized reagent pellets and a paramagnetic stir bar, enabling active mixing through manual rotation of the chip over a stationary magnet. After sample distribution, an oil or air sealing step isolates the chambers, and an integrated reservoir captures excess sample to protect users from biohazard exposure. The device is fabricated from PDMS bonded to glass, with specialized hydrophobic or hydrophilic coatings to regulate fluid flow, and supports both fluorescent and lateral flow detection formats.

This solution is differentiated by its unique combination of high multiplexing capacity, ease of use, and robust reagent management—all without the need for electricity or specialized laboratory equipment. Unlike traditional nucleic acid detection methods that require complex instrumentation or trained personnel, this device leverages CRISPR-based assays (such as Cas13-RPA or SHINE) for high sensitivity and specificity, while its hand-actuated design and pre-stored lyophilized reagents enable true sample-to-answer workflows in resource-limited settings. The hybrid manifold ensures reliable, uniform distribution of sample fluid even at large multiplexing scales, and the innovative magnetic mixing system overcomes the limitations of passive diffusion in microfluidic environments. Its versatility extends from rapid infectious disease diagnostics in the field to laboratory automation, offering a transformative approach to molecular diagnostics that is accessible, scalable, and safe for a wide range of applications.

Benefits

• Enables highly multiplexed nucleic acid detection with up to 100 reaction chambers in a single device.
• Requires no specialized laboratory equipment, supporting point-of-need and field-deployable diagnostics.
• Incorporates lyophilized reagents for long-term, room-temperature storage without refrigeration.
• Utilizes hand-actuated magnetic mixing for rapid and uniform reagent reconstitution, enhancing detection sensitivity.
• Minimizes sample waste and user exposure through integrated fluid containment and sealing mechanisms.
• Supports multiple readout formats including fluorescence and lateral flow for flexible detection options.
• Manufactured using accessible materials and methods, avoiding complex cleanroom processes.
• Provides rapid, sensitive, and specific detection suitable for infectious disease surveillance and outbreak response.

Applications

• Point-of-care infectious disease diagnostics
• Field-deployable outbreak surveillance
• Multiplexed pathogen detection in clinics
• Resource-limited laboratory automation
• Rapid drug-resistance screening

Additional info

This microfluidic platform facilitates 100-plex nucleic acid detection using a hybrid parallel-serial manifold. It employs surface tension-based valves and antechambers for uniform sequential filling via manual injection. Pre-loaded lyophilized reagents and paramagnetic stir bars enable active mixing through manual rotation over a magnet. Fabricated from PDMS-on-glass, the system isolates reaction chambers using oil or air, supporting sensitive fluorescent or lateral flow readouts without laboratory infrastructure.