Self-contained biological assay apparatus, methods, and applications
Inventors
Zhou, Peng • Young, Lincoln C. • Thomas, Benjamin • Chen, Zongyuan • Roswech, Todd • Spizz, Gwendolyn • Yasmin, Rubina • Mouchka, Greg
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9556478-B2
Publication Date
2017-01-31
Expiration Date
Abstract
A self-contained, fully automated, biological assay-performing apparatus includes a housing; a dispensing platform including a controllably-movable reagent dispensing system, disposed in the housing; a reagent supply component disposed in the housing; a pneumatic manifold removably disposed in the housing in a space shared by the dispensing platform, removably coupled to a fluidic transport layer and a plurality of reservoirs, wherein the fluidic transport layer, the reservoirs, and a test sample to be introduced therein are disposed in the housing in the space separate from the dispensing platform; a pneumatic supply system removably coupled to the pneumatic manifold in the housing in a space separate from the dispensing platform; and a control system coupled to at least one of the dispensing platform and the pneumatic supply system, disposed in the housing.
Core Innovation
The invention describes an automated process that uses a self-contained biological assay apparatus with a reagent dispensing system and a pneumatic manifold operating a microfluidic system. The microfluidic system includes a fluidic transport layer with a plurality of fluidic channels and a plurality of reservoirs attached to the fluidic transport layer, all formed in a single microfluidic chip.
A fluid test sample containing a target nucleic acid is introduced into one reservoir that is fluidically connected to a corresponding fluidic channel. At least one liquid reagent is provided from a reservoir to the fluidic channel, and the sample and reagent are combined within the fluidic transport layer, a reservoir, or a temperature-controlled amplification/reaction reactor.
The combined fluid is transported to a temperature-controlled amplification/reaction reactor for incubation under conditions sufficient to amplify the target nucleic acid sequence. The amplified sequence is then transported to an analysis reservoir and analyzed, with the sample automatically transported from a fluid sample introduction reservoir to the analysis reservoir separately from any other samples, and independent of contact with the pneumatic manifold and the reagent dispensing system.
Claims Coverage
The document provides one independent claim defining an automated microfluidic nucleic-acid workflow on a single microfluidic chip with separate sample transport. Dependent claims add inventive refinements in transport and combining mechanisms, microarray-based analysis in a dedicated reservoir, membrane-based fluid routing, optional heating of a membrane, and probe-based interaction detection.
Automated microfluidic nucleic-acid assay with separate sample transport
Automatically transports a fluid test sample from a fluid sample introduction reservoir to an analysis reservoir separately from any other samples, while combining the sample with at least one liquid reagent, incubating in a temperature-controlled amplification/reaction reactor to amplify a target nucleic acid sequence, and analyzing the amplified sequence, all in a single microfluidic chip and independent of contact with the pneumatic manifold and the reagent dispensing system.
Pneumatic manifold with microfluidic channels and reservoirs on a fluidic transport layer
Provides a self-contained biological assay apparatus including a reagent dispensing system and a pneumatic manifold that operates a microfluidic system having a fluidic transport layer with a plurality of fluidic channels and a plurality of reservoirs attached to the fluidic transport layer.
Temperature-controlled amplification/reaction reactor for target nucleic acid amplification
Transports the fluid test sample and the at least one reagent to a temperature-controlled amplification/reaction reactor that is in operative communication with the fluidic transport layer and incubates under conditions sufficient to amplify the target nucleic acid sequence.
Independent analyzing in an analysis reservoir
Transports the fluid test sample and the at least one reagent to an analysis reservoir via a fluidic channel and analyzes the amplified target nucleic acid sequence from the fluid test sample.
Pumping by a multi-valve diaphragm pump controlled by a pneumatic manifold
Transports and combines the fluid test sample and at least one reagent by pumping through the fluidic transport layer using a multi-valve diaphragm pump controlled by the pneumatic manifold.
Microarray analysis in a microarray analysis reservoir
Performs the analyzing step as a microarray analysis in a microarray analysis reservoir within the microfluidic system.
Membrane-based microarray fluid routing with fluid exit along the periphery
Provides a microarray analysis membrane in a reservoir, flows a fluid over the top surface of the microarray analysis membrane, and removes the fluid substantially through a fluid exit route along the periphery of the microarray analysis membrane.
Heating of the microarray analysis membrane
Provides heat to the microarray analysis membrane as part of the microarray analysis process.
Detecting interaction between amplified target sequence and a target-specific probe
Detects an interaction between the amplified target nucleic acid sequence and a probe that is specific to the target nucleic acid sequence.
Overall, the claims cover an automated microfluidic nucleic-acid assay in a self-contained apparatus where sample transport is separate and independent from contact with the pneumatic manifold and reagent dispensing system, followed by temperature-controlled amplification and analysis in an analysis reservoir. Dependent refinements add pneumatic manifold-driven pumping via a multi-valve diaphragm pump, microarray analysis in a dedicated reservoir using a membrane with defined fluid routing, optional membrane heating, and interaction detection between the amplified sequence and a target-specific probe.
Stated Advantages
Documented Applications
Pharmacogenomic warfarin SNP genotyping (CYP2C9, VKORC1) using primer extension/RDB confirmation by sequencing.
Automated multiplex HPV genotyping (20 types) using low-density microarray/RDB with reproducibility and comparative sensitivity to Digene.
Detection of sparse targets (heat-shock mRNA of Cryptosporidium) with immunomagnetic concentration, optional heat shock, amplification (RT-PCR/NASBA), and lateral flow analysis.
Apparatus options including heated analysis reservoirs, pneumatically activated magnet assemblies, and optical real-time PCR modules.
Interested in licensing this patent?