Cartridge for performing assays in a closed sample preparation and reaction system

• US 9,598,722 B2
• Filed: 11/11/2014
• Issued: 03/21/2017
• Est. Priority Date: 11/11/2014
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Genmark Diagnostics Incorporated (Roche Holding AG)
• Inventors: Kroehl, Peter, Kayyem, Jon Faiz, Gray, Darren Scott, Wright, David Walter, Aiello, Dominic
• Primary Examiner(s): WHITE, DENNIS MICHAEL
• Application Number:
  US14/538,533
• Publication Number:
  US 20160130640A1
• Time in Patent Office: 861 Days
• US Class Current: 1/1
• CPC Class Codes: B01F 2101/23 Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
• CPC Class Codes: B01F 27/0721 parallel with respect to the rotating axis
• CPC Class Codes: B01F 27/0723 oblique with respect to the rotating axis
• CPC Class Codes: B01F 27/0724 directly mounted on the rotating axis
• CPC Class Codes: B01F 27/11251 having holes in the surface
• CPC Class Codes: B01F 27/113 Propeller-shaped stirrers for producing an axial flow, e.g. shaped like a ship or aircraft propeller
• CPC Class Codes: B01F 33/304 the mixing being performed in a mixing chamber where the products are brought into contact
• CPC Class Codes: B01F 33/86 Mixing heads comprising a driven stirrer
• CPC Class Codes: B01F 35/2207 Use of data, i.e. barcodes, 3D codes or similar type of tagging information, as instruction or identification codes for controlling the computer programs, e.g. for manipulation, handling, production or compounding in mixing plants B01F33/848 takes precedence
• CPC Class Codes: B01F 35/3204 Motor driven, i.e. by means of an electric or IC motor
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• 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 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• 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 2300/0636 : Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 : Electrodes
• CPC Class Codes: B01L 2300/0816 : Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• 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 2300/161 : Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2300/1822 : using Peltier elements
• CPC Class Codes: B01L 2400/0427 : Electrowetting
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0683 : mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: C12N 15/1006 : by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6825 : Nucleic acid detection involving sensors

Abstract

In one embodiment, a multiplex fluid processing cartridge includes a sample well, a deformable fluid chamber, a mixing well with a mixer disposed therein, a lysis chamber including a lysis mixer, an electrowetting grid for microdroplet manipulation, and electrosensor arrays configured to detect analytes of interest. An instrument for processing the cartridge is configured to receive the cartridge and to selectively apply thermal energy, magnetic force, and electrical connections to one or more discrete locations on the cartridge and is further configured to compress the deformable chamber(s) in a specified sequence.

Claims

1. A fluid sample processing cartridge comprising:

a substrate;

a sample well formed in said substrate and configured to receive a volume of fluid sample;

a closure configured to be selectively placed over said sample well;

a deformable fluid chamber supported on said substrate and configured to hold a fluid therein when in an undeformed state and to collapse upon application of an external compression force to expel at least a portion of the fluid from the fluid chamber, said deformable fluid chamber being in fluid communication with said sample well via a channel formed in said substrate;

a mixing well formed in said substrate, said mixing well being in fluid communication with said sample well via a channel formed in said substrate, said mixing well comprising a first peripheral wall and a first floor defining a well and a fluid inlet snorkel extending up a side of the first peripheral wall extending from the channel communicating the mixing well to the sample well and terminating below a top edge of the first peripheral wall; and

a driven mixing apparatus disposed within said mixing well and constructed and arranged to mix the contents of said mixing well.

2. The fluid sample processing cartridge of claim 1, wherein said fluid inlet snorkel extends up an outer surface of the first peripheral wall and terminates at an opening formed in the first peripheral wall.

3. The fluid sample processing cartridge of claim 1, wherein said sample well comprising a second peripheral wall and a second floor defining a well and a fluid inlet snorkel extending up a side of the second peripheral wall and terminating below a top edge of the second peripheral wall.

4. The fluid sample processing cartridge of claim 1, wherein said mixing well further comprises an exit port comprising one or more openings formed in the second floor, wherein said floor tapers downwardly toward said exit port.

5. The fluid sample processing cartridge of claim 1, wherein said driven mixing apparatus comprises a first impeller rotatably disposed within said mixing well and a gear configured to be drivingly engaged by a mating gear of an instrument into which the liquid sample processing cartridge is inserted and to rotate the first impeller when engaged by the mating gear.

6. The fluid sample processing cartridge of claim 1, further comprising:

a lysis chamber containing a plurality of lysis beads, said lysis chamber being formed in said substrate and disposed along the channel connecting said mixing well and said sample well whereby fluid flowing from the sample well to said mixing well will flow through said lysis chamber; and

a bead mixer disposed at least partially within said lysis chamber and constructed and arranged to agitate the lysis beads and fluid flowing through said lysis chamber.

7. The fluid sample processing cartridge of claim 6, further comprising:

a first optical interface comprising an enlarged portion of the channel connecting said lysis chamber to said sample well; and

a second optical interface comprising an enlarged portion of the channel connecting said lysis chamber to said mixing well.

8. The fluid sample processing cartridge of claim 6, wherein said bead mixer comprises:

a motor mounted within said substrate; and

a second impeller disposed within said lysis chamber and mounted on an output shaft of said motor.

9. The fluid sample processing cartridge of claim 6, wherein said lysis chamber includes a fluid inlet and a fluid outlet and further comprises a mesh filter disposed over each of said fluid inlet and said fluid outlet and configured to retain said lysis beads within said lysis chamber.

10. The fluid sample processing cartridge of claim 1, further comprising:

a pressure port formed in said substrate and configured to couple said substrate to an external fluid pressure source; and

a channel formed in said substrate connecting said pressure port to said sample well.

11. The fluid sample processing cartridge of claim 1, further comprising:

a waste chamber formed in said substrate, said waste chamber being in fluid communication with said mixing well via a channel formed in said substrate;

a fluid exit port formed in the substrate, said fluid exit port being in fluid communication with said mixing well via a channel formed in said substrate;

a first externally actuatable control valve disposed within said substrate and constructed and arranged to selectively permit or prevent fluid flow from said mixing well to said waste chamber; and

a second externally actuatable control valve disposed within said substrate and constructed and arranged to selectively permit or prevent fluid flow from said mixing well to said fluid exit port.

12. The fluid sample processing cartridge of claim 11, further comprising a capture chamber disposed along a channel connecting said mixing well and said waste chamber.

13. The fluid sample processing cartridge of claim 1, further comprising:

a passive valve assembly disposed within said substrate and constructed and arranged to be closed and prevent fluid flow from said mixing well when pressure within said mixing well is not higher than a threshold pressure and to open and permit fluid flow from said mixing well when pressure within said mixing well rises above the threshold pressure; and

a pressure port formed in said substrate and in pressure communication with said passive valve assembly by a pressure conduit formed in said substrate, wherein when said pressure port is closed, pressure within said mixing well is allowed to reach the threshold pressure that will open said passive valve assembly and permit fluid flow from said mixing well, and when said pressure port is open pressure within said mixing cannot not reach the threshold pressure so the passive valve assembly is closed.

14. The fluid sample processing cartridge of claim 1, further comprising a lance blister associated with said deformable fluid chamber;

said lance blister being connected or connectable to the associated deformable fluid chamber and containing a bead retained within the lance blister by a breakable septum, wherein said lance blister is configured to collapse upon application of an external compression force to thereby push the bead through the breakable septum.

15. The fluid sample processing cartridge of claim 1, further comprising an external shroud externally enclosing at least a portion of said cartridge.

16. The fluid sample processing cartridge of claim 1, comprising a plurality of deformable fluid chambers, each of said fluid chambers containing one or more substances selected from the group consisting of a lysis buffer, a wash buffer, an oil, a rehydration buffer, target capture beads, and a binding buffer.

17. The fluid sample processing cartridge of claim 1, further comprising:

a first fluid exit port formed in the substrate, said first fluid exit port being in fluid communication with said mixing well via a channel formed in said substrate;

a second fluid exit port formed in the substrate; and

at least two deformable fluid chambers, one of said two deformable fluid chambers being in fluid communication with said mixing well via a channel formed in said substrate, and the other of said two deformable fluid chambers being in fluid communication with said second fluid exit port via a channel formed in said substrate that is different from the channel communicating the first fluid exit port with said mixing well.

18. The fluid sample processing cartridge of claim 17, wherein the deformable fluid chamber in fluid communication with the mixing well contains a lysis buffer, a wash buffer, target capture beads, or a binding buffer, and the deformable fluid chamber in fluid communication with the second fluid exit contains an oil or a rehydration buffer.