Duplex Reverse Transcription Loop-Mediated Isothermal Amplification on a Nanofluidic Digital Chip (Nano-dChip)

Area of Science:

• Biotechnology
• Nanotechnology
• Infectious Disease Diagnostics

Background:

• Increasing global viral outbreaks necessitate rapid, accessible diagnostic solutions.
• Current diagnostic methods can be time-consuming and expensive, hindering timely outbreak response.

Purpose of the Study:

• To develop a novel nanofluidic digital chip (Nano-dChip) for point-of-care viral RNA detection.
• To achieve rapid, sensitive, specific, and cost-effective detection of viral pathogens.

Main Methods:

• Utilized reverse transcription loop-mediated isothermal amplification (RT-LAMP) for target amplification.
• Integrated compartmentalized reagents within a nanofluidic chip to enhance quantification and minimize contamination.
• Employed a single chip for multiplexed detection of viral RNA.

Main Results:

• Achieved viral RNA detection within 30 minutes at a cost below $0.50 per chip.
• Demonstrated a low detection limit of 10 fM for both SARS-CoV-2 and Influenza H3 RNA.
• Validated the Nano-dChip's capability for highly quantitative and contamination-free detection.

Conclusions:

• The Nano-dChip presents a promising platform for rapid, low-cost, and scalable point-of-care viral RNA diagnostics.
• This technology has significant potential for timely infectious disease outbreak control and public health surveillance.
• The developed diagnostic tool addresses the urgent need for accessible and efficient infectious disease detection methods.