Isothermal nucleic acid amplification reactor with integrated solid state membrane

Technology Overview: 

Researchers in the Bau Lab have developed a disposable microfluidic cassette with an integrated porous extraction flow-through membrane for purifying and concentrating nucleic acids from biological samples. 

• The membrane allows decoupling the sample volume from the reaction volume, enabling relatively large samples to achieve high sensitivity, much greater than is typical for point of care devices.
• The purification step ensures that inhibitors that could interfere with nucleic acid amplification are removed from the sample. 
• An external filament allows for temperature control of the isothermal LAMP-based (loop-mediated isothermal amplification) reaction. 

Products can be detected by monitoring the reaction in real-time with a portable fluorescent reader, a smartphone camera, or visually when a colorimetric dye is used. The utility of the device has been demonstrated by detecting the presence of HIV-1 in oral fluids with a limit of detection of 10 HIV particles.

From Liu et al, 2011.  In (A), a photograph of the single-chamber microfluidic cassette with an integrated membrane.  In (B), the cassette holder is equipped with a thin film heater, a thermocouple, and a seat for the detector.  The fluorescent signal is excited and detected with the portable ESE optical detector in the inset.

Advantages: 

• High detection sensitivity
• Decoupling sample volume from reaction volume
• Portable, cost-effective device
• Readily adaptable to purify nucleic acids from pathogens in food, water, and bodily fluids
• Streamlined identification of genetic identity
• Integration of LAMP with sample-to-assay device
• Rapid detection of reaction products without need for thermocycler or expensive lab equipment

Applications: 

• Point-of-care diagnostics
• Environmental sample testing to monitor level of pathogens in food and water supplies
• Monitor level of infection in individuals

Stage of Development: 

Proof-of-concept testing with HIV detection in non-clinical samples; prototype constructed

Intellectual Property: 

USSN 9,476,102 

Reference Media: 

• Liu C. et al.  Analyst, 2011, 136(10), p. 2069-2076.
• Shih-Chuan L. et al.  Sensors and Actuators B, 2016, 229, p. 232-238.
• Song, J. et al.  Analytical Chemistry, 2016, 88(14), p. 7289–7294.

Desired Partnerships: 

• License
• Co-development

Docket # X5677

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