Apparatus and methods for integrated sample preparation, reaction and detection

• US 8,372,340 B2
• Filed: 02/23/2011
• Issued: 02/12/2013
• Est. Priority Date: 10/19/2005
• Status: Active Grant

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Ching, Jesus, Richardson, Bruce, Bird, Dylan Hilmer, Johnson, Bruce A., Moravick, Keith E.
• Primary Examiner(s): Bullock, In Suk
• Assistant Examiner(s): Pregler, Sharon
• Application Number:
  US13/033,129
• Publication Number:
  US 20110236960A1
• Time in Patent Office: 720 Days
• Field of Search: 435/288.5, 435/206.1, 435/306.1, 435/91.2, 435/287.2, 435/255.6, 435/259, 436/53, 436/177, 436/178, 422500-509, 422/537, 422/539, 422/540, 422/541, 422/542
• US Class Current: 422/63
• 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 25/60 Pump mixers, i.e. mixing within a pump
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 35/712 for feeding fluids
• CPC Class Codes: B01F 35/715 Feeding the components in several steps, e.g. successive steps
• CPC Class Codes: B01F 35/7174 using pistons, plungers or syringes
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• 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 2300/1805 : Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• 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 2400/0622 : distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0633 : with moving parts
• CPC Class Codes: B01L 2400/0644 : rotary valves
• CPC Class Codes: B01L 2400/0683 : mechanically breaking a wall or membrane within a channel or chamber
• 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 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50851 : specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/523 : with means for closing or opening
• CPC Class Codes: B01L 3/527 : for a plurality of reagents
• CPC Class Codes: B01L 7/00 : Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• 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: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 1/38 : Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 2035/0436 : with pre-packaged reagents, i.e. test-packs
• CPC Class Codes: G01N 35/1079 : with means for piercing stoppers or septums
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Cartridges for the isolation of a biological sample and downstream biological assays on the sample are provided. In one embodiment, a nucleic acid sample is isolated from a biological sample and the nucleic acid sample is amplified, for example by the polymerase chain reaction. The cartridges provided herein can also be used for the isolation of non-nucleic acid samples, for example proteins, and to perform downstream reactions on the proteins, for example, binding assays. Instruments for carrying out the downstream biological assays and for detecting the results of the assays are also provided.

Claims

1. An apparatus, comprising:

an isolation module including a first housing and a second housing, the first housing defining a first chamber and a second chamber, at least the first chamber configured to contain a sample, the second housing including a side wall and a puncturable member, the side wall and the puncturable member collectively defining a first volume configured to contain a first substance, a portion of the second housing configured to be disposed within the first housing such that the first volume is in fluid communication with the first chamber when a portion of the puncturable member is punctured; and

a reaction module defining a reaction chamber and a second volume configured to contain a second substance, the reaction module configured to be coupled to the isolation module such that the reaction chamber and the second volume are each in fluid communication with the second chamber of the first housing,

the reaction module including a plunger at least partially disposed within the second volume, the plunger configured exert a force on the second substance to transfer the second substance from the second volume into at least one of the reaction chamber or the second chamber when the plunger is moved within the second volume.

2. The apparatus of claim 1, wherein the reaction module is configured to be removably coupled to the isolation module.

3. The apparatus of claim 1, wherein a portion of the reaction module is disposed within the second chamber of the first housing when the reaction module is coupled to the isolation module.

4. The apparatus of claim 1, wherein:

the reaction module includes a substrate that at least partially defines a first flow path and a second flow path, the first flow path configured to be in fluid communication with the reaction chamber and the second chamber of the first housing, the second flow path configured to be in fluid communication with the second volume and the second chamber of the first housing.

5. The apparatus of claim 1, wherein the isolation module includes a transfer mechanism configured to transfer the sample between the first chamber and the second chamber.

6. The apparatus of claim 1, wherein the isolation module includes a valve at least partially disposed within the first housing, the valve configured to transfer the sample between the first chamber and the second chamber while maintaining fluid isolation between the first chamber and the second chamber.

7. The apparatus of claim 1, wherein the isolation module includes a plunger at least partially disposed within the first volume of the second housing, the plunger of the isolation module configured to puncture the portion of the puncturable member when moved within the first volume.

8. The apparatus of claim 1, wherein the isolation module includes an acoustic coupling member configured to convey ultrasonic energy into the first chamber.

9. The apparatus of claim 1, wherein the first volume contains a first substance comprising a lysis buffer, nucleic acid isolation reagent or a combination thereof.

10. The apparatus of claim 1, wherein the second volume comprises second substance comprising reagents to carry out a PCR.

11. The apparatus of claim 10, wherein the second substance further comprises a reverse transcriptase enzyme.

12. The apparatus of claim 1, wherein the reaction chamber comprises a lyophilized PCR master mix.

13. The apparatus of claim 12, wherein the reaction chamber further comprises a reverse transcriptase enzyme.

14. An apparatus, comprising:

a first module defining a first chamber and a second chamber, the first module including a first transfer mechanism configured to transfer a sample between the first chamber and the second chamber while maintaining fluid isolation between the first chamber and the second chamber;

a second module defining a first volume configured to contain a first substance, a portion of the second module configured to be disposed within the first chamber of the first module when the second module is coupled to the first module such that the first volume is configured to be selectively placed in fluid communication with the first chamber; and

a third module defining a reaction chamber and a second volume, the second volume configured to contain a second substance, the third module configured to be coupled to the first module such that the reaction chamber is in fluid communication with the second chamber and such that the second volume is in fluid communication with the reaction chamber, the third module including a second transfer mechanism and a third transfer mechanism, the second transfer mechanism configured to transfer a portion of the sample between the second chamber and the reaction chamber, the third transfer mechanism configured to transfer a portion of the second substance between the second volume and the reaction chamber.

15. The apparatus of claim 14, wherein a portion of the third module is disposed within the second chamber of the first module when the third module is coupled to the first module.

16. The apparatus of claim 14, wherein the second module includes a puncturable member that defines, at least in part, the first volume, the puncturable member further defining a portion of a boundary of the first chamber of the first module when the second module is coupled to the first module.

17. The apparatus of claim 14, wherein the second module includes a fourth transfer mechanism at least partially disposed within the first volume, the fourth transfer mechanism configured to transfer the first substance from the first volume into the first chamber when the fourth transfer mechanism is actuated.

18. The apparatus of claim 14, wherein:

the first module defines a third chamber between the first chamber and the second chamber; and

the first transfer mechanism includes a valve housing and a valve member, the valve housing defining a first opening in fluid communication with the first chamber and a second opening in fluid communication with the third chamber, the valve member defining a cavity in fluid communication with the first opening when the valve member is in a first position, the cavity in fluid communication with the second opening when the valve member is in a second position.

19. The apparatus of claim 14, wherein the second transfer mechanism includes a plunger disposed within a cylinder in fluid communication with the reaction chamber, the plunger configured to produce a vacuum within the reaction chamber when the plunger is moved within the cylinder.

20. An apparatus, comprising:

a first module including a reaction vial, a substrate and a first transfer mechanism, the reaction vial defining a reaction chamber, the substrate defining at least a portion of a first flow path and a second flow path, the first flow path configured to be in fluid communication with the reaction chamber and an isolation chamber of an isolation module, the first transfer mechanism configured to transfer a sample from the isolation chamber to the reaction chamber when the first transfer mechanism is actuated, the first transfer mechanism including a plunger movably disposed within a housing such that the housing and the plunger define a first volume, the first volume containing a first substance, a portion of the plunger disposed within the first flow path such that the first volume is fluidically isolated from the reaction chamber when the plunger is in a first position within the housing, the portion of the plunger disposed apart from the first flow path such that the first volume is in fluid communication with the reaction chamber when the plunger is in a second position within the housing, the plunger configured to produce a vacuum within the reaction chamber when the plunger is moved from the first position to the second position; and

a second module including a second transfer mechanism and defining a second volume configured to contain a second substance, the second module configured to be coupled to the first module such that the second volume can be selectively placed in fluid communication with the reaction chamber via the second flow path, the second transfer mechanism configured to transfer the second substance from the second volume to the reaction chamber when the second transfer mechanism is actuated.

21. The apparatus of claim 20, wherein:

a portion of the first module is disposed within the isolation chamber of the isolation module when the first module is coupled to the isolation module.

22. The apparatus of claim 20, wherein the plunger is configured to produce the vacuum in the reaction chamber to produce a flow of the sample from the isolation chamber to the reaction chamber.

23. The apparatus of claim 20, wherein the second module includes a puncturable member that defines, at least in part, the second volume, the second volume being placed in fluid communication with the reaction chamber when a portion of the puncturable member is punctured.

24. The apparatus of claim 20, wherein the second transfer mechanism includes a plunger disposed within the second volume of the second module, the plunger of the second transfer mechanism configured to exert a force on the second substance to transfer the second substance from the volume into the reaction chamber when the plunger of the second transfer mechanism is moved within the volume.