Fully 3D printed integrated reactor array for point-of-care molecular diagnostics

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Microfluidics

Background:

• Nucleic acid amplification tests (NAATs) are vital for infectious disease management.
• Current molecular diagnostics often require complex laboratory equipment and trained personnel.
• Point-of-care testing (POCT) is needed for rapid diagnosis in diverse settings.

Purpose of the Study:

• To develop a simple, inexpensive, and disposable 3D printed microfluidic reactor array.
• To enable integrated nucleic acid extraction, concentration, and isothermal amplification.
• To facilitate rapid molecular diagnostic tests for infectious diseases at the point of care.

Main Methods:

• Fabrication of a fully 3D printed microfluidic reactor array.
• Integration of flow-through nucleic acid isolation membranes using a leak-proof polymerization strategy.
• Application of static coating technology for improved biocompatibility.
• Demonstration of both qualitative colorimetric and quantitative fluorescence detection.
• Utilized loop-mediated isothermal amplification (LAMP) for pathogen detection.

Main Results:

• The device successfully performed integrated nucleic acid processing and amplification within 50 minutes.
• Achieved detection limits of 100 fg for Plasmodium falciparum and 50 CFU for Neisseria meningitidis.
• Demonstrated comparable performance to traditional benchtop instruments.
• Showcased suitability for detecting pathogens in various body fluids like plasma and cerebrospinal fluid (CSF).

Conclusions:

• The developed 3D printed microfluidic device offers a rapid, cost-effective solution for molecular diagnostics.
• This technology holds significant potential for point-of-care infectious disease detection in resource-limited environments.
• The multifunctional diagnostic platform enhances accessibility to molecular testing outside traditional laboratory settings.