Device for amplifying target nucleic acid

• US 8,822,183 B2
• Filed: 02/12/2013
• Issued: 09/02/2014
• Est. Priority Date: 04/17/1997
• Status: Expired due to Fees

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America), Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated), The United States of America As Represented By The Secretary of Agriculture
• Inventors: Brown, James F., Silver, Jonathan E.
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US13/764,970
• Publication Number:
  US 20130143218A1
• Time in Patent Office: 567 Days
• Field of Search: None
• US Class Current: 435/91.2
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/06 Fluid handling related problems
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• 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 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• 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 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• 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 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A device for amplifying target nucleic acid in a sample can include a planar fluidic assembly including a transparent substrate, a porous material layer on a surface of the transparent substrate, and a cover over the porous material layer and sealingly affixed to the substrate. The cover may be spaced from the porous material layer and a flow channel defined between the porous material layer and the cover. The flow channel may have a uniform cross-section from a first end to a second end. The assembly can further include an inlet in flow communication with the first end of the flow channel to introduce sample containing target nucleic acid into the flow channel, an outlet in flow communication with the second end of the flow channel, and a plurality of nucleic acid primers retained by the porous material layer at discrete regions along and within the flow channel, each of the plurality of nucleic acid primers being complementary to a portion of the target nucleic acid in the sample to enable a primer-based amplification reaction of the target nucleic acid. The porous material layer may be configured to retain, at the discrete regions and during a primer-based amplification reaction, sample introduced to the flow channel and amplified product of the amplification reaction.

Claims

1. A device for amplifying target nucleic acid in a sample, the device comprising:

a planar fluidic assembly comprising;

a substrate,

a porous material layer on a surface of the substrate,

a cover over the porous material layer and sealingly affixed to the substrate, wherein the cover is spaced from the porous material layer;

a flow channel defined between the porous material layer and the cover, wherein the flow channel has a substantially uniform cross-section from a first end to a second end,

an inlet in flow communication with the first end of the flow channel to introduce sample containing target nucleic acid into the flow channel,

an outlet in flow communication with the second end of the flow channel,

a plurality of nucleic acid primers retained by the porous material layer at discrete regions along and within the flow channel, each of the plurality of nucleic acid primers being complementary to a portion of the target nucleic acid in the sample to enable a primer-based amplification reaction of the target nucleic acid,

wherein the porous material layer is configured to retain, at the discrete regions and during the primer-based amplification reaction, the sample introduced to the flow channel and amplified product of the amplification reaction.

2. The device of claim 1, further comprising fluorescence-based detection reagents retained by the porous material layer at the discrete regions.

3. The device of claim 1, wherein the flow channel comprises a plurality of flow channels.

4. The device of claim 1, wherein the porous material layer comprises a plurality of individual porous patches located at the discrete regions.

5. The device of claim 1, wherein the discrete regions are arranged in a two-dimensional array.

6. The device of claim 1, wherein the cover is spaced at a substantially uniform distance from the porous material layer.