On-chip Nanoplasmonic RT-RPA and CRISPR/Cas12a Assay for Point-of-care Molecular Diagnostics

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Nanotechnology

Background:

• Accurate nucleic acid detection at the point-of-care (POC) is crucial for disease diagnosis.
• Existing methods can be time-consuming and require complex laboratory equipment.

Purpose of the Study:

• To develop a compact, all-in-one nanoplasmonic device for rapid on-chip nucleic acid detection.
• To integrate recombinase polymerase amplification (RPA) and CRISPR/Cas12a detection for real-time fluorescence monitoring.

Main Methods:

• Utilized a nanoplasmonic cavity (AuNIs-NC) for efficient photothermal heating.
• Employed a disposable plastic-on-polymer (PoP) cartridge for integrated reactions.
• Integrated a fluorescence microlens array (FMLA) camera for signal detection.

Main Results:

• Demonstrated rapid detection of SARS-CoV-2 E gene plasmid DNA within 20 minutes.
• Achieved a sensitive detection limit of 10 copies/µL.
• Clinical sample testing showed a linear trend with RT-qPCR, confirming reliability.

Conclusions:

• The developed platform provides affordable and reliable molecular diagnostics at POC.
• This integrated plasmonic system facilitates rapid and scalable testing for infectious diseases.
• The on-chip assay offers precise molecular diagnostics for effective disease management.