Systems and methods for sampling of amplification products
Inventors
Thorne, IV, Edward H. • Esterman, Abbie L.
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9689029-B2
Publication Date
2017-06-27
Expiration Date
Abstract
The invention provides systems and methods for processing samples. In a method, a reaction card is provided that has a channel network, a valve, and a micropump, all disposed within the card. The reaction card also has a collection well disposed on a surface of the card and a tubular member extending out from the card. A reaction vessel is provided and affixed to the reaction card such that the tubular member is inserted into the reaction vessel. Amplification reaction reagents and a sample are delivered into the reaction vessel, and an amplification reaction is initiated within the reaction vessel, resulting in an amplification product being disposed within the reaction vessel. The valve is opened to atmosphere, and the first micropump is activated to pump an aliquot of reaction product from the reaction vessel into the tubular member, through the channel network, and into the collection well.
Core Innovation
A nucleic-acid amplification product is sampled after an amplification reaction by using a microfluidic reaction card that includes a channel network, one or more valve(s), and one or more micropump(s). The reaction card further includes a collection well formed in and open to a first surface, and a first tubular member extending out from a second surface opposite the first surface. The valve and micropump are operably connected with the channel network, and the first tubular member and collection well are in fluid communication with the channel network.
The reaction card is coupled to a reaction vessel assembly that comprises a reaction vessel. The reaction vessel assembly is affixed to the reaction card on the second surface so that the first tubular member is inserted into the reaction vessel. The valve is configured to open the reaction vessel to atmosphere, enabling processing of fluids connected to the channel network, including directing amplification products into the collection well.
The system supports electrophoretic analysis by aspirating an aliquot of amplification product from the collection well into an analysis device, detector, and processor. Multiple reaction-card configurations are disclosed, including configurations that manage waste and rinse handling. Real-time quantitative PCR is supported through repeated aliquot sampling during PCR cycles, and end-point workflows are also described, including managing unused product and carryover/cross-contamination.
Claims Coverage
The independent claim is clm-00001. It recites three main inventive groupings: a reaction card architecture with channel network, valve, and micropump coupled to a collection well and a first tubular member; a reaction vessel assembly affixed to the reaction card such that the tubular member inserts into the reaction vessel; and a valve operable to open the reaction vessel to atmosphere. Dependent claims add an analysis subsystem and specific analysis as electrophoretic separation, plus additional waste and handling structures and interface and attachment refinements.
Microfluidic reaction card with channel network, valve, and micropump
A reaction card comprising a channel network, a valve, and a micropump, wherein the valve and the micropump are operably connected with the channel network.
Collection well and tubular member in fluid communication with the channel network
A collection well formed in and open to a first surface of the reaction card, and a first tubular member extending out from a second surface opposite the first surface, wherein the first tubular member and collection well are in fluid communication with the channel network.
Affixed reaction vessel assembly with inserted tubular member
A reaction vessel assembly comprising a reaction vessel, wherein the reaction vessel assembly is affixed to the reaction card on the second surface such that the first tubular member is inserted into the reaction vessel.
Valve operable to open the reaction vessel to atmosphere
The reaction vessel assembly is configured such that the valve is operable to open the reaction vessel to atmosphere.
Integrated analysis device, detector, and processor
The system includes an analysis device, a detector connected to the analysis device, and a processor coupled to the reaction card and the analysis device to perform analysis.
Electrophoretic separation performed by the analysis device
The system is configured so the analysis device performs electrophoretic separation.
Waste well connected to the channel network
The reaction card further includes a waste well formed open to a first surface and connected in fluid communication to the channel network.
Sealing means permitting passage of a tubular member
The system includes a sealing means that allows a first tubular member to pass through when the reaction vessel assembly is attached to the reaction card.
Reagents for performing an amplification reaction
The system includes one or more reagents that are reagents for performing an amplification reaction.
Across the independent claim and its dependent refinements, the core system combines a microfluidic channel network with a valve and micropump on a reaction card, a collection well and a first tubular member in fluid communication with the channel network, and an affixed reaction vessel assembly in which the tubular member inserts into the reaction vessel. The valve is operable to open the reaction vessel to atmosphere, and the disclosed refinements integrate analysis, including electrophoretic separation, along with waste and sealing structures and reagents for an amplification reaction.
Stated Advantages
Documented Applications
No documented applications found
Interested in licensing this patent?