MESOSCALE POLYNUCLEOTIDE AMPLIFICATION DEVICES

• US 20110020876A1
• Filed: 08/11/2003
• Published: 01/27/2011
• Est. Priority Date: 05/01/1992
• Status: Active Grant

• Current Assignee: Larry J. Kricka, Peter Wilding
• Original Assignee: Larry J. Kricka, Peter Wilding
• Inventors: Kricka, Larry J., Wilding, Peter
• Granted Patent: US 7,892,819 B2
• Time in Patent Office: Days
• US Class Current: 435/91.2
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 67/0058 by selective elimination of components, e.g. by leaching
• CPC Class Codes: B01D 67/0072 by deposition from the gaseous phase, e.g. sputtering, CVD, PVD
• CPC Class Codes: B01D 71/02 Inorganic material
• CPC Class Codes: B01F 25/41 Mixers of the fractal type
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• 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 2200/146 Employing pressure sensors
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• 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 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• 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 3/502753 : 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: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12M 41/46 : of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: Y10S 366/03 : Micromixers: variable geometry from the pathway influences mixing/agitation of non-laminar fluid flow
• CPC Class Codes: Y10S 435/808 : Optical sensing apparatus
• CPC Class Codes: Y10S 435/81 : Packaged device or kit
• CPC Class Codes: Y10S 435/814 : Enzyme separation or purification
• CPC Class Codes: Y10T 436/25125 : Digestion or removing interfering materials

Abstract

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction, chamber, having at least one cross-sectional dimension of about 0.1 to 1000 μm. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.

Claims

1-44. -44. (canceled)

45. A device for amplifying a polynucleotide in a sample by conducting a polynucleotide amplification reaction, the device comprising a solid substrate microfabricated to define:

a sample inlet port;

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

a polynucleotide amplification region of the device, in fluid communication with said flow channel, said region having at least one cross-sectional dimension of width or depth of about 0.1 to 1000 microns, at least said polynucleotide amplification region being permanently sealed with a cover; and

an appliance for use in combination with said substrate, said appliance including a nesting site for receiving said substrate and a conduit in fluid communication with the inlet port of said substrate.

46. The device of claim 45 wherein the polynucleotide amplification region comprises a reaction chamber which has at least one dissimilar cross-sectional dimension from a cross-sectional dimension of the sample flow channel.

47. The device of claim 45 wherein said polynucleotide amplification region is configured to contain at least one reagent for conducting a polymerase chain reaction.

48. The device of claim 45 wherein said polynucleotide amplification region is configured to contain at least one reagent for conducting a ligase chain reaction.

49. The device of claim 45 wherein said polynucleotide amplification region is configured to contain at least one reagent for conducting an isothermal DNA amplification reaction.

50. The device of claim 45 wherein said polynucleotide amplification region of the device has at least one cross-sectional dimension of width or depth of about 0.1 to about 500 microns.

51. The device of claim 45 wherein said polynucleotide amplification region of the device has at least one cross-sectional dimension of width or depth of about 0.1 to about 100 microns.

52. The device of claim 45 wherein said polynucleotide amplification region of the device has at least one cross-sectional dimension of width or depth of about 2 to about 50 microns.

53. The device of claim 45 further including means for thermally cycling the contents of said amplification region whereby in each cycle the temperature is controlled to dehybridize double stranded polynucleotide and to permit the in vitro amplification of the sample polynucleotide.

58. The device of claim 45, further comprising a pump actuable to cause fluid flow through the flow system of said substrate.

59. A device of claim 45, wherein said pump is disposed in said appliance and connected to said conduit.

60. The device of claim 53, wherein said means for thermally cycling comprises an electromagnetic energy source, said electromagnetic energy source being provided in said appliance.

54. A method for amplifying a polynucleotide in a sample by conducting a polynucleotide amplification reaction, the method comprising:

(i) providing a device comprising

a solid substrate microfabricated to define;

a sample inlet port;

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

a polynucleotide amplification region, in fluid communication with said flow channel, said region having at least one cross sectional dimension of width or depth of about 0.1 to 1000 microns, at least said polynucleotide amplification region being permanently sealed with a cover; and

an appliance for use in combination with said substrate, said appliance including a nesting site for receiving said substrate and a conduit in fluid communication with the inlet port of said substrate,

(ii) delivering through said conduit, said inlet port and said sample flow channel to said polynucleotide amplification region, said sample polynucleotide and at least one reagent required for the polynucleotide amplification reaction; and

(iii) thermally controlling the contents of the amplification region to amplify said polynucleotide.

55. The method of claim 54 wherein said delivering comprises delivering at least one reagent for conducting a polymerase chain reaction.

56. The method of claim 54 wherein said delivering comprises delivering at least one reagent for conducting a ligase chain reaction.

57. The method of claim 54 wherein said delivering comprises delivering at least one reagent for conducting an isothermal DNA amplification reaction.