Microfluidic cartridge for processing and detecting nucleic acids

• US 9,403,165 B2
• Filed: 06/30/2015
• Issued: 08/02/2016
• 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, Kusner, Michael T.
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US14/755,821
• Publication Number:
  US 20150298119A1
• Time in Patent Office: 399 Days
• Field of Search: None
• US Class Current: 1/1
• 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 for processing a sample, the cartridge comprising:

a first layer comprising a sample port and a fluid port at a broad surface of the first layer;

an intermediate substrate coupled to the first layer and partially separated from the first layer by a film layer, and configured to form a sealed waste chamber with a corrugated surface directly opposing the first layer, wherein the corrugated surface defines a void external to the waste chamber; and

a fluidic pathway, superior to the intermediate substrate and at least partially separated from the corrugated surface of the waste chamber by an elastomeric layer, wherein the fluidic pathway is fluidly coupled to the sample port and the fluid port, and wherein, upon deformation of the elastomeric layer at an occlusion position of the fluidic pathway, by way of the void of the corrugated surface, the fluidic pathway is configured to transfer waste fluid through an opening of the intermediate substrate and into an interior portion of the corrugated surface of the waste chamber.

2. The cartridge of claim 1, wherein the first layer is a unitary construction comprising a reagent port, the fluid port, and a detection chamber, and wherein the fluidic pathway is coupled to the reagent port, the fluid port, and the detection chamber.

3. The cartridge of claim 1, wherein the waste chamber comprises at least one waste inlet in communication with the opening of the intermediate substrate, and wherein the corrugated surface of the waste chamber further defines a rectangular prismatic void spanning a long dimension of the cartridge that defines a magnet housing region configured to reversibly receive a magnet from the direction perpendicular to a broad face of the first layer, and configured to cross the fluidic pathway.

4. The cartridge of claim 3, wherein the fluidic pathway comprises a capture segment, downstream of the sample port and configured to cross the magnet housing region multiple times.

5. The cartridge of claim 4, wherein the fluidic pathway is configured to transfer waste fluid of the sample to the waste chamber through a first waste inlet upstream of the capture segment by way of a first waste segment fluidly coupled to the first waste inlet and an initiating portion of the capture segment, and wherein the fluidic pathway is configured to transfer a second waste fluid to the waste chamber through a second waste inlet downstream of the capture segment by way of a second waste segment fluidly coupled to the second waste inlet and a terminating portion of the capture segment.

6. The cartridge of claim 1, wherein the elastomeric layer is situated between the fluidic pathway and the intermediate substrate, wherein the intermediate substrate provides access to a set of occlusion positions of the fluidic pathway through at least at a subset of a set of voids of the corrugated surface of the waste chamber, such that the fluidic pathway is configured to be occluded through the elastomeric layer at the set of occlusion positions from a direction perpendicular to the broad surface of the first layer.

7. The cartridge of claim 6, wherein at least one void of the set of voids defined by the corrugated surface of the waste chamber is defined by five surfaces of the corrugated surface, including a first pair of parallel surfaces and a second pair of parallel surfaces orthogonal to the first pair of parallel surfaces and orthogonal to the broad surface of the first layer, and a surface, coupled to the first pair of surfaces and the second pair of surfaces and proximal to the first layer, having a set of openings for occlusion of the fluidic pathway at the set of occlusion positions, and wherein occluding the fluidic pathway at a first subset of the set of occlusion positions defines a truncated pathway configured to facilitate transfer of waste fluid of the sample to the waste chamber.

8. A cartridge for processing a sample, the cartridge comprising:

a first layer and an intermediate substrate coupled to the first layer and partially separated from the first layer by a film layer, wherein the intermediate substrate is configured to form a sealed waste chamber with a corrugated surface directly opposing the first layer, wherein the corrugated surface defines a set of parallel voids external to the waste chamber; and

a first fluidic pathway, formed by at least a portion of the first layer; and

a second fluidic pathway in parallel with the first fluidic pathway and formed by at least a portion of the second fluidic pathway, wherein the first fluidic pathway and the second fluidic pathway are each at least partially separated from the corrugated surface by an elastomeric layer, and each fluidic pathway is configured to transfer waste fluid of the sample into the waste chamber through a set of openings of the intermediate substrate.

9. The cartridge of claim 8, wherein the elastomeric layer is inferior to the film layer.

10. The cartridge of claim 8, wherein the first layer is a unitary construction comprising a first sample port-reagent port pair including a first sample port, a second sample port-reagent port pair including a second sample port, a fluid port, a first detection chamber, and a second detection chamber, wherein the first fluidic pathway is coupled to the first sample port-reagent port pair and the first detection chamber, wherein the second fluidic pathway is coupled to the second sample port-reagent port pair and the second detection chamber, and wherein at least one of the first fluidic pathway and the second fluidic pathway is coupled to the fluid port.

11. The cartridge of claim 10, further comprising 1) a heating region defined as a recessed region of the first layer that is parallel to the set of voids of the corrugated surface, and 2) a vent 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.

12. The cartridge of claim 10, wherein the fluid port is fluidly coupled to the first fluidic pathway and to the second fluidic pathway.

13. The cartridge of claim 9, wherein the first detection chamber comprises a first serpentine-shaped fluidic channel, and the second detection chamber comprises a second serpentine-shaped fluidic channel.

14. The cartridge of claim 13, wherein the first serpentine-shaped fluidic channel comprises three wide channels directly interconnected by two narrow channels, wherein the three wide channels include a first wide channel comprising a detection chamber inlet into the first detection chamber and a second wide channel comprising a detection chamber outlet from the first detection chamber.

15. The cartridge of claim 8, wherein at least one void of the set of voids defined by the corrugated surface of the waste chamber is a rectangular prismatic void spanning a long dimension of the cartridge and defining a magnet housing region configured to cross the first fluidic pathway and the second fluidic pathway.

16. The cartridge of claim 8, wherein the elastomeric layer is situated between the first layer and the intermediate substrate, wherein the intermediate substrate provides access to a set of occlusion positions of the first fluidic pathway and of the second fluidic pathway by way of the set of voids of the corrugated surface of the waste chamber, such that the first fluidic pathway and the second fluidic pathway are configured to be occluded upon deformation of the elastomeric layer at subsets of the set of occlusion positions, and wherein the waste chamber is located inferior to the elastomeric layer.

17. The cartridge of claim 16, wherein the first fluidic pathway is configured to transfer a first waste fluid of the sample to the waste chamber through a first waste inlet, upon occlusion of a first subset of the set of occlusion positions, by way of a first waste segment fluidly coupled to the first waste inlet and a first portion of the first fluidic pathway proximal the first sample port, and wherein the first fluidic pathway is configured to transfer a second waste fluid of the sample to the waste chamber through a second waste inlet, upon occlusion of a second subset of the set of occlusion positions, by way of a second waste segment fluidly coupled to the second waste inlet and a second portion of the first fluidic pathway substantially downstream of the first sample port.

18. The cartridge of claim 16, wherein the first fluidic pathway is configured to transfer a first waste fluid to the waste chamber through a first waste inlet, upon occlusion of a first subset of the set of occlusion positions, by way of a first waste segment fluidly coupled to the first waste inlet and inline with the first fluidic pathway, and wherein the second fluidic pathway is configured to transfer a second waste fluid to the waste chamber through a second waste inlet, upon occlusion of a second subset of the set of occlusion positions, by way of a second waste segment fluidly coupled to the second waste inlet and inline with the second fluidic pathway.