An automated, fully-integrated nucleic acid analyzer based on microfluidic liquid handling robot technique

Area of Science:

• Biotechnology
• Medical Diagnostics
• Analytical Chemistry

Background:

• On-site nucleic acid testing (NAT) is crucial for timely disease monitoring and pathogen identification.
• Existing NAT methods can be complex and require specialized laboratory settings.
• There is a need for integrated, automated, and cost-effective solutions for point-of-care diagnostics.

Purpose of the Study:

• To develop an automated and fully-integrated nucleic acid analyzer.
• To combine automated liquid handling with microfluidic droplet-based real-time PCR.
• To create a compact, low-cost system for on-site pathogen detection.

Main Methods:

• Developed an integrated system combining automated liquid handling robots and microfluidic droplet-based real-time PCR.
• Incorporated automated sample introduction, magnetic solid-phase extraction for nucleic acid extraction, reverse transcription, and droplet generation.
• Utilized a simple chip for reagent pre-loading and oil-covered PCR reaction droplets, enabling real-time dual fluorescence detection.

Main Results:

• Achieved a compact analyzer size (50 × 45 × 45 cm) and a low instrument cost (approx. $900).
• Demonstrated a low single analysis cost (less than $5).
• Successfully identified eight types of influenza pathogens in human throat swabs with results consistent with the colloidal gold method.

Conclusions:

• The developed automated nucleic acid analyzer offers a compact, low-cost, and efficient solution for on-site pathogen detection.
• The integrated system streamlines complex NAT procedures, making them accessible outside traditional labs.
• This technology has significant potential for disease monitoring and rapid diagnostics in various settings.