Integrated nucleic acid diagnostic device and method for in-situ confocal microscopy
First Claim
Patent Images
1. A miniature analytical device, comprising:
- a body having a plurality of distinct reaction chambers disposed therein, each of said reaction chambers being fluidly connected to at least one other of said reaction chambers;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device;
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers; and
a hybridization chamber for analyzing a component of said fluid sample, said hybridization chamber being fluidly connected to at least one of said plurality of reaction chambers and including a polymer array, said polymer array including a plurality of different polymer sequences coupled to a surface of a single substrate, each of said plurality of different polymer sequences being coupled to said surface in a different, known location.
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Abstract
The present invention provides a miniaturized integrated nucleic acid diagnostic device and system. The device of the invention is generally capable of performing one or more sample acquisition and preparation operations, in combination with one or more sample analysis operations. For example, the device can integrate several or all of the operations involved in sample acquisition and storage, sample preparation and sample analysis, within a single integrated unit. The device is useful in a variety of applications, and most notably, nucleic acid based diagnostic applications and de novo sequencing applications.
71 Citations
79 Claims
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1. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein, each of said reaction chambers being fluidly connected to at least one other of said reaction chambers;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device;
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers; and
a hybridization chamber for analyzing a component of said fluid sample, said hybridization chamber being fluidly connected to at least one of said plurality of reaction chambers and including a polymer array, said polymer array including a plurality of different polymer sequences coupled to a surface of a single substrate, each of said plurality of different polymer sequences being coupled to said surface in a different, known location. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein, each of said reaction chambers being fluidly connected to at least one other of said reaction chambers;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device;
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers; and
a microcapillary channel for analyzing a component of said fluid sample, said microcapillary channel being fluidly connected to at least one of said plurality of reaction chambers and including at least first and second electrodes at opposite ends of said microcapillary channel for applying a voltage across said microcapillary channel. - View Dependent Claims (19, 20, 21, 22, 24)
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23. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein, at least one of said reaction chambers being an in vitro transcription reaction chamber, said in vitro transcription reaction chamber having an effective amount of an RNA polymerase and four different nucleoside triphosphates, disposed therein;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device; and
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers. - View Dependent Claims (25)
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26. A miniature analytical device, comprising:
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a body having at least first, second and third reaction chambers disposed within said body;
said first reaction chamber having an opening disposed through said body for introducing a fluid sample into said first reaction chamber, said second reaction chamber having disposed therein at least one reagent for amplifying a nucleic acid segment within a sample, and said third reaction chamber having an array of oligonucleotides disposed therein, said array including a plurality of different nucleic acid sequences coupled to a surface of a single substrate, each of said plurality of different nucleic acid sequences being coupled to said surface in a different, known location, and disposed within said third chamber for hybridizing with at least a portion of a nucleic acid segment amplified in said second reaction chamber; and
a fluid transport system for transporting a fluid sample from said first reaction chamber to said second reaction chamber and from said second reaction chamber to said third reaction chamber. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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37. A miniature analytical device, comprising:
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a body having at least three distinct reaction chambers disposed therein, wherein each of said reaction chambers is fluidly connected to at least one other of said reaction chambers;
a first of said reaction chambers including a cell lysis system disposed therein, for lysing cells in a fluid sample;
a second of said reaction chambers having amplification reagents disposed therein, for amplifying a nucleic acid derived from said cells lysed in said first reaction chamber;
a third reaction chamber having an oligonucleotide array disposed therein, said oligonucleotide array including a plurality of different nucleic acid sequences coupled to a surface of a single substrate, each of said plurality of different nucleic acid sequences being coupled to said surface in a different, known location;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device; and
a fluid transport system for moving a fluid sample from said at least first reaction chamber to said at least second reaction chamber, and from said at least second reaction chamber to said at least third reaction chamber. - View Dependent Claims (38, 39, 40, 41, 42, 43, 44)
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45. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein, each of said reaction chambers being fluidly connected to at least one other of said reaction chambers, at least one of said reaction chambers being a temperature controlled reaction chamber having a controllable heating element disposed therein;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device; and
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52, 53, 54)
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55. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein, each of said reaction chambers being fluidly connected to at least one other of said reaction chambers;
a fluid mixing system for generating convection within at least one of said reaction chambers;
a sample inlet, fluidly connected to at least one of said plurality of reaction chambers, for introducing a fluid sample into said device; and
a fluid transport system for moving a fluid sample from at least a first reaction chamber of said plurality of reaction chambers to at least a second reaction chamber of said plurality of reaction chambers. - View Dependent Claims (56, 57, 58, 59, 60, 61, 62, 63)
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64. A miniature analytical device, comprising:
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a body having a plurality of distinct reaction chambers disposed therein;
a sample inlet, fluidly connected to at least one of said plurality of distinct reaction chambers, for introducing a fluid sample into said device;
a central pumping chamber disposed within said body, said central pumping chamber being fluidly connected to each of said plurality of reaction chambers by one of a plurality of fluid passages, each of said plurality of fluid passages including a valve disposed across said fluid passage, whereby said fluid passages may be selectively opened and claosed to direct a fluid sample from a first of said plurality of reaction chambers through said central pumping chamber and into a second of said plurality of reaction chambers. - View Dependent Claims (65, 66, 67, 68, 70)
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69. A miniature analytical device, comprising:
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a body having at least a first reaction chamber fluidly connected to a second reaction chamber by a fluid passage;
a sample inlet, fluidly connected to said first reaction chamber, for introducing a fluid sample into said device;
a differential pressure delivery system for maintaining said first reaction chamber at a first pressure and said second reaction chamber at a second pressure, said first pressure being greater than ambient pressure and said second pressure being greater than said first pressure, whereby when said second reaction chamber is brought to ambient pressure, said first pressure forces a liquid sample in said first reaction chamber into said second reaction chamber. - View Dependent Claims (71, 72, 73)
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74. A miniature analytical device, comprising:
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a body having at least a first reaction chamber fluidly connected to a second reaction chamber;
a sample inlet, fluidly connected to said first reaction chamber, for introducing a fluid sample into said device;
a differential pressure delivery source for maintaining said first reaction chamber at a first pressure and said second reaction chamber at a second pressure, said second pressure being less than ambient pressure and said first pressure being less than said second pressure, whereby when said first reaction chamber is brought to ambient pressure, said second pressure draws a liquid sample in said first reaction chamber into said second reaction chamber. - View Dependent Claims (75, 76, 77, 78, 79)
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Specification