Microfluidic cartridge for processing and detecting nucleic acids

• US 9,738,887 B2
• Filed: 02/13/2013
• Issued: 08/22/2017
• Est. Priority Date: 02/13/2012
• Status: Active Grant

• Current Assignee: Neumodx Molecular, Inc. (Qiagen NV)
• Original Assignee: Neumodx Molecular, Inc. (Qiagen NV)
• Inventors: Williams, Jeffrey, Brahmasandra, Sundaresh, Kusner, Michael T.
• Primary Examiner(s): Hobbs, Michael
• Assistant Examiner(s): HASSAN, LIBAN M
• Application Number:
  US13/765,996
• Publication Number:
  US 20130210125A1
• Time in Patent Office: 1,651 Days
• Field of Search: 4352872
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0615 Loss of fluid by dripping
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/022 Transponder chips
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0609 Holders integrated in container to position an object
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/0816 : Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 : Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 : comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 : Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 : Multiple sequential chambers
• CPC Class Codes: B01L 2300/0883 : Serpentine channels
• CPC Class Codes: B01L 2300/0887 : Laminated structure
• CPC Class Codes: B01L 2300/0893 : having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 2300/123 : Flexible; Elastomeric
• CPC Class Codes: B01L 2300/14 : Means for pressure control
• CPC Class Codes: B01L 2300/16 : Surface properties and coatings
• CPC Class Codes: B01L 2300/1805 : Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1822 : using Peltier elements
• CPC Class Codes: B01L 2300/1827 : using resistive heater
• CPC Class Codes: B01L 2400/0406 : capillary forces
• CPC Class Codes: B01L 2400/043 : magnetic forces
• CPC Class Codes: B01L 2400/0478 : pistons
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0622 : distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/065 : sliding valves
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/0694 : vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 2400/086 : using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502 : with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 : characterised by venting arrangements
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/508 : rigid containers not provided for above
• CPC Class Codes: B01L 3/527 : for a plurality of reagents
• CPC Class Codes: B01L 3/567 : Valves, taps or stop-cocks in combination with burettes B01L3/0203; in general F16K
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B29C 65/08 : using ultrasonic vibrations non-plastics element to plastics elements B29C65/645
• CPC Class Codes: B29C 65/484 : Moisture curing adhesives
• CPC Class Codes: B29C 65/606 : the rivets being integral with one of the parts to be joined, i.e. staking
• CPC Class Codes: B29C 66/71 : characterised by the composition of the plastics material of the parts to be joined welding bar compositions B29C65/125
• CPC Class Codes: B29C 66/81423 : being concave
• CPC Class Codes: B29C 66/8322 : Joining or pressing tools reciprocating along one axis
• CPC Class Codes: B29K 2021/00 : Use of unspecified rubbers as moulding material
• CPC Class Codes: B29L 2031/756 : Microarticles, nanoarticles
• CPC Class Codes: C12M 23/42 : Integrated assemblies, e.g. cassettes or cartridges
• CPC Class Codes: C12N 13/00 : Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 15/1013 : by using magnetic beads
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6813 : Hybridisation assays
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]

Abstract

A microfluidic cartridge, configured to facilitate processing and detection of nucleic acids, comprising: a top layer comprising a set of cartridge-aligning indentations, a set of sample port-reagent port pairs, a shared fluid port, a vent region, a heating region, and a set of Detection chambers; an intermediate substrate, coupled to the top layer comprising a waste chamber; an elastomeric layer, partially situated on the intermediate substrate; and a set of fluidic pathways, each formed by at least a portion of the top layer and a portion of the elastomeric layer, wherein each fluidic pathway is fluidically coupled to a sample port-reagent port pair, the shared fluid port, and a Detection chamber, comprises a turnabout portion passing through the heating region, and is configured to be occluded upon deformation of the elastomeric layer, to transfer a waste fluid to the waste chamber, and to pass through the vent region.

Claims

1. A cartridge, configured to facilitate processing and detecting of a nucleic acid, comprising:

a first layer, defining a sample port, a reagent port, a fluid port, and a detection chamber;

an elastomeric layer;

an intermediate substrate coupled to the first layer, such that the elastomeric layer is situated between the intermediate substrate and the first layer, wherein the intermediate substrate defines a chamber with a corrugated surface directly opposing the first layer, wherein the corrugated surface defines a set of voids external to the chamber and accessible from a direction perpendicular to a broad surface of the first layer, and wherein at least a portion of the corrugated surface includes a set of openings that provide access to the elastomeric layer; and

a fluidic pathway, wherein the fluidic pathway is fluidically coupled to the sample port, the reagent port, the fluid port, and the detection chamber.

2. The cartridge of claim 1

wherein the fluidic pathway is formed by at least a portion of the first layer and a portion of the elastomeric layer, is configured to be occluded upon manipulation of the elastomeric layer through the set of openings of the corrugated surface, and is configured to transfer a waste fluid to the chamber.

3. The cartridge of claim 2, wherein the intermediate substrate is partially separated from the first layer by a film layer.

4. The cartridge of claim 2, wherein the chamber of the corrugated surface includes a waste inlet coupled to the fluidic pathway and a waste vent situated at a first side of the fluidic pathway, and wherein the cartridge further comprises a vent region directly opposed to the waste vent at a second side of the fluidic pathway.

5. The cartridge of claim 4, wherein a void of the set of voids of the corrugated surface of the chamber receives and positions a magnet proximal the fluidic pathway during operation.

6. The cartridge of claim 5, wherein the fluidic pathway comprises a turnabout segment directly adjacent to the void and that crosses the void.

7. The cartridge of claim 2, wherein the set of openings of the corrugated surface of the intermediate substrate define a set of valve guides that guide occluding objects toward the elastomeric layer at a set of occlusion positions, during operation.

8. The cartridge of claim 1, further comprising a vent region, a region configured to receive a magnet, and a heating region, such that a segment of the fluidic pathway is configured to cross the region configured to receive the magnet and the heating region, and the fluidic pathway is configured to pass through the vent region upstream of the detection chamber.

9. The cartridge of claim 8, wherein the heating region is defined by a recessed region of the first layer and spans a long dimension of the first layer.

10. The cartridge of claim 1, wherein a terminal portion of the fluidic pathway is coupled to an end vent, configured to provide fine metering of fluid flow.

11. The cartridge of claim 1, wherein the detection chamber comprises a first, a second, and a third detection chamber segment wherein each of the first, the second, and the third detection chamber segment is a broad chamber of which a projection onto a plane is substantially rectangular, wherein a first end of the second detection chamber segment is connected to the first detection chamber segment by a first narrow fluidic channel, and wherein a second end of the second detection chamber segment is connected to the third detection chamber segment by a second narrow fluidic channel.

12. A cartridge, configured to facilitate processing and detecting of a nucleic acid, comprising:

a first layer comprising a sample port and a detection chamber;

an elastomeric layer;

an intermediate substrate including a set of valve guides, wherein the intermediate substrate defines a chamber with a corrugated surface directly opposing the first layer, wherein the corrugated surface defines a set of voids external to the chamber and accessible from a direction perpendicular to a broad surface of the first layer, and wherein at least a portion of the corrugated surface defines the set of valve guides with a set of openings that provide access to the elastomeric layer; and

a fluidic pathway, formed by at least a portion of the first layer and a portion of the elastomeric layer, wherein the fluidic pathway is fluidically coupled to the sample port and the detection chamber and comprises a first and second branch extending downstream from a junction, and is configured to be occluded at a set of occlusion positions upon manipulation of the elastomeric layer through the set of valve guides, wherein a first occlusion position of the set of occlusion positions is positioned along the fluidic pathway downstream of the junction and upstream of the first branch and a second occlusion position of the set of occlusion positions is positioned along the fluidic pathway downstream of the junction and upstream of the second branch, wherein the set of occlusion positions comprises a normally open position and a normally closed position,

wherein the normally open position comprises a first surface of the fluidic pathway at the first layer and a second surface of the fluidic pathway at the elastomeric layer, wherein a void defined between the first surface and the second surface is configured to transition to a closed state upon occlusion by an occluding object applied to the elastomeric layer during operation;

wherein the normally closed position is defined by a region of the fluidic pathway, at the first layer that extends toward and abuts the elastomeric layer in preventing fluid bypass at the region;

wherein a first truncated pathway, including the normally open position and the first branch and excluding the second branch, is defined upon manipulation of the fluidic pathway at the first and second occlusion positions, and

wherein a second truncated pathway, including the normally closed position and the second branch and excluding the first branch, to the detection chamber is defined upon manipulation of the fluidic pathway at the first and second occlusion positions.

13. The cartridge of claim 12, wherein the first layer further comprises a reagent port and a fluid port, and wherein the sample port and the reagent port are configured to couple to a standard pipette tip, and the fluid port is configured to couple to a syringe pump.

14. The cartridge of claim 12, wherein the first layer further comprises a vent region including a liquid-impermeable membrane, and wherein the fluidic pathway is configured to pass through the vent region upstream of the detection chamber.

15. The cartridge of claim 12, wherein the corrugated surface defines a region configured to receive a magnet providing a magnetic field during operation.

16. The cartridge of claim 15, wherein the fluidic pathway comprises a turnabout segment configured to repeatedly cross the region configured to receive a magnet.

17. The cartridge of claim 12, wherein a third truncated pathway, coupled to the fluid port, not coupled to the sample port, and not coupled to the reagent port, is defined upon occlusion of the fluidic pathway at a third subset of the set occlusion positions.

18. The cartridge of claim 17, wherein the first subset of the set of occlusion positions, the second subset of the set of occlusion positions, and the third subset of the set of occlusion positions are overlapping.

24. The cartridge of claim 12, wherein the normally closed position is configured to prevent fluid bypass, and, during operation of the cartridge, is configured to be forced open due to fluid pressure applied by a fluid delivery system.

19. A cartridge, configured to facilitate processing and detecting nucleic acids, comprising:

a shared fluid port configured to receive a processing reagent volume;

a first fluidic pathway for processing a first sample;

a second fluidic pathway for processing a second sample;

wherein the first fluidic pathway is coupled to the shared fluid port and the second fluidic pathway is coupled to the shared fluid port, wherein the first fluidic pathway and the second fluidic pathway are each configured to be occluded at a set of occlusion positions, and wherein the set of occlusion positions comprises a normally open position and a normally closed position,

wherein the normally open position comprises a first surface of the fluidic pathway at the first layer and a second surface of the fluidic pathway at the elastomeric layer, wherein a void defined between the first surface and the second surface is configured to transition to a closed state upon occlusion by an occluding object applied to the elastomeric layer during operation;

wherein the normally closed position is defined by a region of the fluidic pathway, at the first layer that extends toward and abuts the elastomeric layer in preventing fluid bypass at the region; and

a first layer, an elastomeric layer, and an intermediate substrate that defines a chamber with a corrugated surface directly opposing the first layer, wherein the corrugated surface defines a set of voids external to the chamber and accessible from a direction perpendicular to a broad surface of the first layer, and wherein at least a portion of the corrugated surface includes a set of openings that provide access to the elastomeric layer.

20. The cartridge of claim 19,

wherein the first layer defines a first sample port-reagent port pair, a second sample port-reagent port pair, the shared fluid port, a first detection chamber, and a second detection chamber,

wherein the elastomeric layer is situated between the intermediate substrate and the first layer,

wherein the first fluidic pathway is formed by at least a portion of the first layer and a portion of the elastomeric layer, and is coupled to the first sample port-reagent port pair and to the first detection chamber, and

wherein the second fluidic pathway is formed by at least a portion of the first layer and a portion of the elastomeric layer, and is coupled to the second sample port-reagent port pair and to the second detection chamber.

21. The cartridge of claim 20, wherein the first layer further comprises a vent region and a heating region, such that the first fluidic pathway is configured to cross the heating region and to pass through the vent region upstream of the first detection chamber, and the second fluidic pathway is configured to cross the heating region and to pass through the vent region upstream of the second detection chamber.

22. The cartridge of claim 19, wherein the corrugated surface of the chamber partially forms a region configured to receive a magnet, which substantially spans a long dimension of the microfluidic cartridge to cross the first fluidic pathway and the second fluidic pathway.

23. The cartridge of claim 19, wherein the first fluidic pathway and the second fluidic pathway are identical.