Microchip module for blood sample preparation and nucleic acid amplification reactions.

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Diagnostics

Background:

• Nucleic acid amplification reactions and sample preparation are crucial for genetic testing.
• Integrating these steps into a single microchip platform can streamline diagnostic procedures.

Purpose of the Study:

• To design and construct a microchip module for the integrated isolation of white blood cells and subsequent polymerase chain reaction (PCR) amplification.
• To demonstrate the feasibility of a microchip-based system for simplified nucleic acid analyses.

Main Methods:

• A computer numerical control-machined plexiglas microchip module was developed.
• The module incorporates a custom heater-cooler for thermal cycling and microchannels for reagent transport.
• White blood cells were isolated using "weir-type" filters within an 8–9 µL dual-purpose glass-silicon microchip.
• A 226-bp region of the human coagulation Factor V gene was amplified via microchip-based PCR.

Main Results:

• Successful isolation of white blood cells from <3 µL of human whole blood.
• Direct amplification of a specific genomic target (Factor V gene) from isolated white blood cells on the microchip.
• Demonstration of integrated cell isolation and PCR within the microchip module.

Conclusions:

• The developed microchip module effectively integrates blood sample preparation (cell isolation) and nucleic acid amplification (PCR).
• This integrated system offers a simplified and convenient approach for nucleic acid analyses.
• The platform shows potential for adaptation to various integrated molecular assays.