A 3D-printed, self-driven microfluidic sensor chip for point-of-care testing (POCT) of norovirus

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Diagnostics

Background:

• Early norovirus (NV) diagnosis is vital for outbreak control.
• CRISPR-coupled isothermal amplification offers sensitive nucleic acid detection but faces multi-enzyme compatibility challenges.

Purpose of the Study:

• To develop a microfluidic sensor for cascade isothermal amplification and CRISPR-mediated reactions.
• To integrate sample pretreatment and glucose biosensing for NV detection.
• To create a point-of-care testing (POCT) system for NV diagnostics.

Main Methods:

• A 3D-printed microfluidic device was designed for integrated sample pretreatment and cascade amplification.
• Reverse transcription-recombinase polymerase amplification (RT-RPA) coupled with CRISPR was employed.
• A personal glucose meter (PGM) was utilized for digital quantification of NV nucleic acids.

Main Results:

• The system achieved high sensitivity and specificity in detecting NV nucleic acids.
• Linear correlation observed between log NV target RNA concentration and fluorescence intensity.
• The microfluidic chip-based POCT system demonstrated a limit of detection of approximately 60 copies of NV.

Conclusions:

• The developed microfluidic sensor enables sensitive and specific NV detection.
• The integrated RT-RPA-CRISPR assay is suitable for POCT applications.
• This approach facilitates field-deployable diagnostics for viral pathogens.