Microfluidic cartridge for processing and detecting nucleic acids

• US 10,557,132 B2
• Filed: 01/16/2017
• Issued: 02/11/2020
• 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 S., Brahmasandra, Sundaresh, Kusner, Michael T.
• Primary Examiner(s): Hassan, Liban M
• Application Number:
  US15/407,160
• Publication Number:
  US 20170120243A1
• Time in Patent Office: 1,121 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 for processing a sample, the cartridge comprising:

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

an intermediate substrate coupled to the first layer and defining a chamber opposing the first layer, wherein the chamber comprises an external surface defining a set of voids configured to receive a set of pins;

an actuation layer between the first layer and the intermediate substrate; and

a fluidic pathway defined by at least a portion of the first layer and at least partially separated from the chamber by the actuation layer, the fluidic pathway fluidly coupled to the sample port and the fluid port, and configured to transport fluid through the cartridge upon manipulation of the actuation layer by the set of pins; and

a set of chamber inlets configured to deliver fluid from the fluidic pathway into the chamber, wherein the chamber is fluidly accessible at more than one location along the fluidic pathway through the set of chamber inlets, and wherein the cartridge is configured to drive fluid from the fluidic pathway into the chamber, through at least a subset of the set of chamber inlets, upon manipulation of the actuation layer by the set of pins.

2. The cartridge of claim 1, wherein the intermediate substrate is partially separated from the first layer by a film layer.

3. The cartridge of claim 1, wherein the actuation layer is an elastomeric layer comprising a set of regions operable to restrict flow through the fluidic pathway upon deformation by the set of one or more pins.

4. The cartridge of claim 1, wherein the actuation layer and the set of voids of the chamber of the intermediate substrate form a set of valves operable to affect flow through the fluidic pathway.

5. The cartridge of claim 1, wherein the intermediate substrate comprises a rectangular prismatic void defined by the external surface of the chamber of the intermediate substrate that spans a length of the cartridge and defines a magnet housing region operable to reversibly receive a magnet.

6. The cartridge of claim 5, wherein the fluidic pathway comprises an s-shaped capture segment, downstream of the sample port and configured to cross the magnet housing region multiple times.

7. The cartridge of claim 6, wherein the fluidic pathway comprises a detection chamber downstream of the s-shaped capture segment, and wherein the cartridge is operable to drive fluid from the fluidic pathway into the detection chamber, upon manipulation of the actuation layer.

8. The cartridge of claim 1, wherein the fluid port comprises an interface to a fluid handling system comprising a pump.

9. A cartridge for processing of a sample, the cartridge comprising:

a first layer;

an intermediate substrate coupled to the first layer and defining an internal chamber, wherein the internal chamber comprises an external surface defining a set of voids configured to receive a set of pins;

an actuation layer between the first layer and the intermediate substrate;

a first fluidic pathway, formed by at least a portion of the first layer and a portion of the actuation layer, and having a first sample inlet;

a set of internal chamber inlets; and

a second fluidic pathway in parallel with the first fluidic pathway, formed by at least a portion of the first layer and a portion of the actuation layer, and having a second sample inlet, wherein the first fluidic pathway and the second fluidic pathway share a fluid inlet and are each configured to transport fluid from the fluid inlet into the internal chamber through the set of internal chamber inlets, the set of internal chamber inlets defined by the intermediate substrate, wherein the internal chamber is fluidly accessible at more than one location along the first and second fluidic pathways through the set of internal chamber inlets, wherein the cartridge is configured to drive fluid from the first and second fluidic pathways into the internal chamber, through at least a subset of the set of internal chamber inlets, upon manipulation of the actuation layer by the set of pins.

10. The cartridge of claim 9, wherein the actuation layer is accessible through the set of voids of the intermediate substrate.

11. The cartridge of claim 10, wherein the actuation layer is an elastomeric layer comprising a set of regions configured to restrict flow through at least one of the first fluidic pathway and the second fluidic pathway upon deformation through the set of voids.

12. The cartridge of claim 11, 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, such that the first fluidic pathway and the second fluidic pathway are operable to be occluded upon deformation of the elastomeric layer at subsets of the set of occlusion positions.

13. The cartridge of claim 10, wherein the actuation layer and the set of voids of the intermediate substrate form a set of valves operable to affect flow through at least one of the first fluidic pathway and the second fluidic pathway.

14. The cartridge of claim 9, further comprising 1) a heating region defined as a recessed region of the first layer, 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 a first detection chamber, and the second fluidic pathway is configured to cross the heating region and to pass through the vent region upstream of a second detection chamber.

15. The cartridge of claim 14, wherein the first detection chamber comprises a first serpentine-shaped fluidic channel, and the second detection chamber comprises a second serpentine-shaped fluidic channel.

16. The cartridge of claim 9, wherein the fluid inlet comprises an interface to a fluid handling system comprising a pump.