Integrated microfluidic device and methods
Inventors
Zhou, Peng • Young, Lincoln C. • Roswech, Todd • Spizz, Gwendolyn • Chen, Zongyuan • Thomas, Benjamin W. • Lee, Travis
Assignees
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Publication Number
US-9132398-B2
Publication Date
2015-09-15
Expiration Date
Abstract
A microfluidic device for analyzing a sample of interest is provided. The microfluidic device can comprise a microfluidic device body, wherein the microfluidic device body comprises a sample preparation area, a nucleic acid amplification area, a nucleic acid analysis area, and a network of fluid channels. Each of the sample preparation area, the nucleic acid amplification area and the nucleic acid analysis area are fluidly interconnected to at least one of the other two areas by at least one of the fluid channels. Using the microfluidic device, sample preparation can be combined with amplification of a biologically active molecule, and a suitable biological sample can be provided for analysis and/or detection of a molecule of interest. The small-scale apparatus and methods provided are easier, faster, less expensive, and equally efficacious compared to larger scale equipment for the preparation and analysis of a biological sample.
Core Innovation
The invention provides a microfluidic device for analyzing a sample of interest. The microfluidic device body comprises a sample preparation area, a non-actuated, fixed volume nucleic acid amplification area, and a plurality of fluid channels interconnected in a network, with the sample preparation area and the nucleic acid amplification area fluidly interconnected to the other area by at least one of the plurality of fluid channels.
A single, non-elastomeric substrate layer with upper and lower surfaces and one or more microfeatures in the upper surface is contacted and joined with a single, non-elastomeric membrane layer disposed over the one or more microfeatures to form a diaphragm valve. In a relaxed state the membrane lies substantially against the upper surface, and in an actuated state the membrane is moved away from the upper surface.
The document further describes integrating sample preparation, nucleic acid amplification, and nucleic acid analysis areas via the network of fluid channels to enable automated end-point nucleic acid assays with minimal user handling. Pressure control is implemented using differential pressure sources and diaphragm/valve structures, and the disclosed workflow supports probe-based interaction assays, including reverse dot blot (RDB), with image and threshold-based signal quantification.
Claims Coverage
The independent claim is clm-00001. It centers on core device architecture features, with dependent claim refinements adding pressure-to-valve actuation, sample preparation subcomponents, defined amplification-area reservoirs, silica membrane purification media, and threshold-based detection.
Interconnected microfluidic sample preparation and fixed-volume amplification areas
A microfluidic device body including a sample preparation area, a non-actuated, fixed volume nucleic acid amplification area, and a plurality of fluid channels interconnected in a network, where the sample preparation area and the nucleic acid amplification area are fluidly interconnected via at least one of the plurality of fluid channels.
Two-layer non-elastomeric diaphragm valve construction with relaxed and actuated membrane states
A single, non-elastomeric substrate layer with upper and lower surfaces and one or more microfeatures, and a single, non-elastomeric membrane layer contacting and joined with the upper surface and disposed over the microfeatures to form a diaphragm valve, with a relaxed state where the membrane lies substantially against the upper surface and an actuated state where the membrane is moved away from the upper surface.
Differential pressure transformed by diaphragms positioned in multiple channels
At least one diaphragm disposed in at least two of the plurality of fluid channels to transform pressure from a differential pressure source to a desired open or closed position.
Sample preparation area with fluidly interconnected intake reservoir, reagent reservoir, and purification media
The sample preparation area comprising a sample intake reservoir, a reservoir for a sample preparation reagent, and sample purification media, with the three being fluidly interconnected.
Nucleic acid amplification area with reactor and interconnected reservoirs
The nucleic acid amplification area including a nucleic acid amplification reactor, a nucleic acid amplification reagent reservoir, and a nucleic acid amplification product reservoir fluidly interconnected.
Silica membrane as sample purification media
The sample purification media is a silica membrane.
Threshold-based detection by comparing color intensity to a computed threshold value
Computing a threshold value and comparing a color intensity value to the threshold value to detect the molecule of interest.
The coverage centers on an integrated microfluidic architecture with interconnected sample preparation and fixed-volume nucleic acid amplification areas, combined with a two-layer non-elastomeric diaphragm valve that changes between relaxed and actuated membrane states. Additional refinements specify pressure-to-valve state transformation, detailed sample preparation and amplification-area reservoir architectures, silica membrane purification media, and threshold-based detection.
Stated Advantages
Automated end-point nucleic acid assays with minimal user handling.
Comparable performance to bench-top extraction/PCR, including time reduction and equivalence across sample types.
Documented Applications
Disease/disorder detection, including targets and examples such as HPV, HIV, and β-thalassemia.
Detection in sample matrices including E. coli in food matrices.
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