Integrated fluid manipulation cartridge

• US 6,440,725 B1
• Filed: 06/25/1999
• Issued: 08/27/2002
• Est. Priority Date: 12/24/1997
• Status: Expired due to Term

• Current Assignee: Cepheid (Danaher Corporation)
• Original Assignee: Cepheid (Danaher Corporation)
• Inventors: Kovacs, Gregory T. A., Chang, Ronald, Petersen, Kurt E., Northrup, M. Allen, McMillan, William A., Pourahmadi, Farzad, Christel, Lee A., Ching, Jesus
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US09/331,911
• Time in Patent Office: 1,159 Days
• Field of Search: 435/287.2, 435/287.3, 435/287.9, 435/288.5, 435/288.6, 435/306.1, 422/58-61, 422/68.1, 422/70, 422/99, 422/101, 422/104, 422/102, 241/2
• US Class Current: 435/288.5
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01L 2200/0621 Control of the sequence of chambers filled or emptied
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• 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 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/046 Chemical or electrochemical formation of bubbles
• 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 2400/0644 rotary valves
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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: C12M 47/02 Separating microorganisms from the culture medium; Concentration of biomass separating microorganisms from their culture media C12N1/02
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01F 1/708 : Measuring the time taken to traverse a fixed distance
• CPC Class Codes: G01N 30/6091 : Cartridges
• CPC Class Codes: G01N 30/6095 : Micromachined or nanomachined, e.g. micro- or nanosize
• 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

A cartridge (101) for separating a desired analyte from a fluid sample includes a sample port (103) and a sample flow path extending from the port through the body of the cartridge. The sample flow path includes at least one flow-through component (122), e.g., filter paper or a microfabricated chip, for capturing the desired analyte from the sample as the sample flows through the cartridge. The cartridge also includes an elution flow path for carrying elution fluid through the component (122) to release captured analyte from the component into the elution fluid. The elution flow path diverges from the sample flow path after passing through the component (122). Flow controllers (41A) and (41B) direct the remaining fluid sample into a waste chamber (139) after the sample flows through the component (122) and direct the elution fluid and eluted analyte into a reagent chamber (141) and reaction chamber (143). The continuous-flow design of the cartridge permits the rapid processing of a fluid sample that is larger in volume than any interactive region within the cartridge, allowing increased sensitivity in the detection of low copy concentrations of analytes, such as nucleic acid.

Claims

1. A device for use with an ultrasonic transducer to separate an analyte from a fluid sample, the device comprising a cartridge having:

a) a first flow path including;

i) a lysing chamber for lysing sample components to release the analyte therefrom, wherein the lysing chamber contains a membrane or filter for capturing the sample components as the sample flows through the lysing chamber, and wherein the lysing chamber is defined by at least one wall for contacting the transducer to effect the transfer of ultrasonic energy to the lysing chamber;

ii) an extraction chamber, downstream of the lysing chamber, the extraction chamber containing at least one solid support for capturing the released analyte; and

iii) at least a first waste chamber, downstream of the extraction chamber, for receiving waste fluid;

b) a second flow path for eluting the captured analyte from the extraction chamber, wherein the second flow path passes through the extraction chamber and diverges from the first flow path after passing through the extraction chamber; and

c) at least one flow controller for directing the waste fluid into the waste chamber and for directing the eluted analyte into the second flow path.

2. The device of claim 1, further comprising beads in the lysing chamber for rupturing the sample components.

3. The device of claim 1, wherein the at least one solid support in the extraction chamber is selected from the group consisting of filters, beads, fibers, membranes, glass wool, filter paper, polymers and gels.

4. The device of claim 1, wherein the cartridge further includes a reagent chamber in fluid communication with the lysing chamber for holding a lysing reagent.

5. The device of claim 1, further comprising:

a second waste chamber, fluidicly connected to the lysing chamber, for receiving waste fluid from the lysing chamber; and

flow control means for directing the waste fluid from the lysing chamber into the second waste chamber and for directing the analyte released in the lysing chamber into the extraction chamber.

6. The device of claim 1, further comprising a heating element for heating the extraction chamber, thereby improving elution efficiency.

7. The device of claim 1, wherein the cartridge further includes a reagent chamber in fluid communication with the extraction chamber via the second flow path, the reagent chamber containing dried reagents.

8. The device of claim 1, wherein the cartridge further includes a reaction chamber, in fluid communication with the extraction chamber via the second flow path, for holding the analyte for amplification or detection.

9. The device of claim 8, wherein the reaction chamber contains dried reagents.

10. The device of claim 1, wherein the cartridge further includes:

a reagent chamber in fluid communication with the extraction chamber via the second flow path, the reagent chamber containing dried reagents; and

a reaction chamber, in fluid communication with the reagent chamber, for holding the analyte for amplification or detection.

11. The device of claim 1, wherein the cartridge includes a sample port for introducing the sample into the cartridge.

12. The device of claim 11, wherein the sample port comprises a receiving area for receiving the sample.

13. The device of claim 12, further comprising a syringe or needle for moving the sample into the receiving area of the cartridge.

14. The device of claim 1, wherein the cartridge further includes a storage chamber positioned at the beginning of the second flow path for holding an elution fluid.

15. The device of claim 1, further comprising a reaction vessel coupled to the cartridge for receiving the eluted analyte via the second flow path.

16. The device of claim 1, further comprising an external instrument for receiving the cartridge, wherein the external instrument includes a fluid motive source for forcing fluids to flow in the cartridge.

17. The device of claim 1, wherein the cartridge includes an integrated fluid motive source for forcing fluids to flow within the cartridge.

18. The device of claim 17, wherein the fluid motive source comprises at least one membrane pump.

19. The device of claim 11, wherein the wall comprises a polymeric material, and wherein the wall has a thickness in the range of 0.01 to 0.5 mm.

20. The device of claim 1, wherein the extraction chamber is formed in a microfoluidic chip enclosed within the body of the cartridge, and wherein the chip has inlet and outlet ports in fluid communication with the extraction chamber, the inlet and outlet ports being positioned to permit continuous fluid flow through the extraction chamber.

21. A device for use with an ultrasonic transducer to separate an analyte from a fluid sample containing cells, spores, or microorganisms, the device comprising a cartridge having:

a) a lysing chamber for lysing the cells, spores, or microorganisms to produce a lysate containing the analyte, wherein the lysing chamber contains a membrane or filter for capturing the cells, spores, or microorganisms as the sample flows through the lysing chamber, and wherein the lysing chamber is defined by at least one wall for contacting the transducer to effect the transfer of ultrasonic energy to the lysing chamber;

b) an extraction chamber for receiving the lysate, the extraction chamber containing at least one solid support for capturing the analyte in the lysate as the lysate flows through the extraction chamber;

c) an elution flow path passing through the extraction chamber for eluting the captured analyte from the extraction chamber;

d) at least one waste chamber for receiving waste fluid from the lysing and extraction chambers; and

e) flow control means for;

i) directing the sample to flow through the lysing chamber and into the at least one waste chamber, thereby capturing the sample components to be lysed with the membrane or filter in the lysing chamber;

ii) directing the lysate to flow from the lysing chamber through the extraction chamber and into the at least one waste chamber, thereby capturing the analyte in the lysate with the at least one solid support in the extraction chamber; and

iii) directing an elution fluid to flow through the extraction chamber and along the elution flow path, thereby eluting the analyte from the extraction chamber.

22. The device of claim 21, further comprising beads in the lysing chamber for rupturing the cells, spores, or microorganisms.

23. The device of claim 21, wherein the at least one solid support in the extraction chamber is selected from the group consisting of filters, beads, fibers, membranes, glass wool, filter paper, polymers and gels.

24. The device of claim 21, wherein the cartridge further includes a reagent chamber in fluid communication with the lysing chamber for holding a lysing reagent.

25. The device of claim 21, wherein the cartridge includes a first waste chamber for receiving waste fluid from the lysing chamber and a second waste chamber for receiving waste fluid from the extraction chamber.

26. The device of claim 21, further comprising a heating element for heating the extraction chamber, thereby improving elution efficiency.

27. The device of claim 21, wherein the cartridge includes a sample port for introducing the sample into the cartridge.

28. The device of claim 27, wherein the sample port comprises a receiving area for receiving the sample.

29. The device of claim 28, further comprising a syringe or needle for moving the sample into the receiving area of the cartridge.

30. The device of claim 21, wherein the cartridge further includes a reagent chamber in fluid communication with the extraction chamber via the elution flow path, the reagent chamber containing dried reagents.

31. The device of claim 21, wherein the cartridge further includes a reaction chamber, in fluid communication with the extraction chamber via the elution flow path, for holding the analyte for amplification or detection.

32. The device of claim 31, wherein the reaction chamber contains dried reagents.

33. The device of claim 21, wherein the cartridge further includes:

a reagent chamber in fluid communication with the extraction chamber via the elution flow path, the reagent chamber containing dried reagents to be mixed with the analyte; and

a reaction chamber, in fluid communication with the reagent chamber, for holding the analyte for amplification or detection.

34. The device of claim 21, wherein the cartridge further includes a storage chamber positioned at the beginning of the elution flow path for holding an elution fluid.

35. The device of claim 21, further comprising a reaction vessel coupled to the cartridge for receiving the eluted analyte.

36. The device of claim 21, wherein the wall comprises a polymeric material, and wherein the wall has a thickness in the range of 0.01 to 0.5 mm.

37. A device for separating nucleic acid from a fluid sample and for holding the nucleic acid for an amplification reaction, the device comprising:

a) a cartridge having;

i) a first flow path including;

an extraction chamber containing at least one solid support for capturing the nucleic acid; and

a waste chamber, downstream of the extraction chamber, for receiving waste fluid;

ii) a second flow path for eluting the captured nucleic acid from the extraction chamber, wherein the second flow path passes through the extraction chamber and diverges from the first flow path after passing through the extraction chamber; and

iii) at least one flow controller for directing the waste fluid into the waste chamber and for directing the eluted nucleic acid into the second flow path; and

b) a reaction vessel coupled to the cartridge for receiving the eluted nucleic acid, the reaction vessel having a reaction chamber for holding the nucleic acid for the amplification reaction, and the reaction vessel being coupled to the cartridge such that the vessel protrudes from the body of the cartridge.

38. The device of claim 37, further comprising an external instrument for receiving the cartridge, wherein the external instrument includes a fluid motive source for forcing fluids to flow in the cartridge.

39. The device of claim 37, wherein the cartridge includes an integrated fluid motive source for forcing fluids to flow within the cartridge.

40. The device of claim 39, wherein the fluid motive source comprises at least one membrane pump.

41. The device of claim 37, wherein the at least one solid support in the extraction chamber is selected from the group consisting of filters, beads, fibers, membranes, glass wool, filter paper, polymers and gels.

42. The device of claim 37, wherein the extraction chamber is formed in a microfluidic chip enclosed within the body of the cartridge, and wherein the at least one solid support comprises an array of microstructures extending into the extraction chamber, each of the microstructures having an aspect ratio of at least 2:

1.

43. The device of claim 37, further comprising a heating element for heating the extraction chamber, thereby improving elution efficiency.

44. The device of claim 37, wherein the cartridge further includes a lysing region f or lysing sample components to release the nucleic acid therefrom.

45. The device of claim 44, wherein the cartridge further includes a reagent chamber in fluid communication with the lysing region for holding a lysing reagent.

46. The device of claim 44, wherein the lysing region contains a membrane or filter for capturing the sample components as the sample flows through the lysing region, and wherein the lysing region is defined by at least one wall for contacting an ultrasonic transducer to effect the transfer of ultrasonic energy to the lysing region.

47. The device of claim 46, further comprising beads in the lysing region for rupturing the sample components.

48. The device of claim 46, wherein the wall comprises a polymeric material, and wherein the wall has a thickness in the range of 0.01 to 0.5 mm.

49. The device of claim 37, where in the reaction vessel is fluidicly connected to the extraction chamber via the second flow path.

50. The device of claim 37, wherein the cartridge further includes a reagent chamber fluidicly connected to the extraction chamber via the second flow path, the reagent chamber containing dried reagents for mixing with the eluted nucleic acid, and wherein the reaction vessel is fluidicly connected to the reagent chamber to receive the eluted nucleic acid and reagents.

51. The device of claim 37, wherein the reaction vessel contains dried reagents.

52. The device of claim 37, wherein the cartridge includes a sample port for introducing the sample into the cartridge.

53. The device of claim 52, wherein the sample port comprises a receiving area for receiving the sample.

54. The device of claim 53, further comprising a syringe or needle for moving the sample into the receiving area of the cartridge.

55. The device of claim 37, wherein the cartridge further includes a storage chamber positioned at the beginning of the second flow path for holding an elution fluid.

56. A device for separating nucleic acid from a fluid sample and for holding the nucleic acid for an amplification reaction, the device comprising a cartridge having:

a) a first flow path including;

i) an extraction chamber containing at least one solid support for capturing the nucleic acid; and

ii) a waste chamber, downstream of the extraction chamber, for receiving waste fluid;

b) a second flow path for eluting the captured nucleic acid from the extraction chamber, wherein the second flow path passes through the extraction chamber and diverges from the first flow path after passing through the extraction chamber;

c) at least one flow controller for directing the waste fluid into the waste chamber and for directing the eluted nucleic acid into the second flow path; and

d) a reaction chamber for receiving the eluted nucleic acid via the second flow path and for holding the nucleic acid for the amplification reaction, wherein the portion of the cartridge defining the reaction chamber protrudes from the rest of the cartridge body.

57. The device of claim 56, further comprising an external instrument for receiving the cartridge, wherein the external instrument includes a fluid motive source for forcing fluids to flow in the cartridge.

58. The device of claim 56, wherein the cartridge includes an integrated fluid motive source for forcing fluids to flow within the cartridge.

59. The device of claim 58, wherein the fluid motive source comprises at least one membrane pump.

60. The device of claim 56, wherein the at least one solid support in the extraction chamber is selected from the group consisting of filters, beads, fibers, membranes, glass wool, filter paper, polymers and gels.

61. The device of claim 56, wherein the extraction chamber is formed in a microfluidic chip enclosed within the body of the cartridge, and wherein the at least one solid support comprises an array of microstructures extending into the extraction chamber, each of the microstructures having an aspect ratio of at least 2:

1.

62. The device of claim 56, further comprising a heating element for heating the extraction chamber, thereby improving elution efficiency.

63. The device of claim 56, wherein the cartridge further includes a lysing region for lysing sample components to release the nucleic acid therefrom.

64. The device of claim 63, wherein the cartridge further includes a reagent chamber in fluid communication with the lysing region for holding a lysing reagent.

65. The device of claim 63, wherein the lysing region contains a membrane or filter for capturing the sample components as the sample flows through the lysing region, and wherein the lysing region is defined by at least one wall for contacting an ultrasonic transducer to effect the transfer of ultrasonic energy to the lysing region.

66. The device of claim 65, further comprising beads in the lysing region for rupturing the sample components.

67. The device of claim 65, wherein the wall comprises a polymeric material, and wherein the wall has a thickness in the range of 0.01 to 0.5 mm.

68. The device of claim 56, wherein the cartridge further includes a reagent chamber in the second flow path, the reagent chamber containing dried reagents.

69. The device of claim 56, wherein the reaction chamber contains dried reagents.

70. The device of claim 56, wherein the cartridge includes a sample port for introducing the sample into the cartridge.

71. The device of claim 70, wherein the sample port comprises a receiving area for receiving the sample.

72. The device of claim 71, further comprising a syringe or needle for moving the sample into the receiving area of the cartridge.

73. The device of claim 56, wherein the cartridge further includes a storage chamber positioned at the beginning of the second flow path for holding an elution fluid.