Device for amplifying and detecting a target nucleic acid

• US 7,582,470 B2
• Filed: 07/17/2003
• Issued: 09/01/2009
• Est. Priority Date: 07/31/2002
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Browne, Kenneth A.
• Primary Examiner(s): Strzelecka; Teresa E
• Application Number:
  US10/621,803
• Publication Number:
  US 20040023284A1
• Time in Patent Office: 2,238 Days
• Field of Search: 435/174, 435/283.1, 422/50, 422/68.1, 422/82.05
• US Class Current: 435/287.1
• 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 2219/00313 the reactor vessels being formed by arrays of wells in blocks
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C40B 30/04 by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/14 Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 for creating libraries

Abstract

Compositions, methods and devices for detecting nucleic acids. The invention particularly regards composite arrays of immobilized amplification primers and hybridization probes. Also disclosed are compositions and methods for covalently immobilizing oligonucleotides and other biological molecules to glass and plastic surfaces.

Claims

1. A device for amplifying and detecting a target nucleic acid, comprising:

a solid support bead having a surface;

an amplification primer immobilized to the surface of said solid support bead, said amplification primer comprising a promoter sequence for an RNA polymerase and a sequence complementary to a first strand of said target nucleic acid; and

a labeled hybridization probe separate from the amplification primer immobilized to said surface,

wherein said labeled hybridization probe comprises 2′

-methoxy nucleotide analogs and a sequence complementary to an amplicon synthesized using said amplification primer and said target nucleic acid as a template in a nucleic acid amplification reaction, and

wherein prior to contact of said device with any nucleotide polymerizing enzyme said labeled hybridization probe comprises a detectable label and is immobilized to said surface.

2. The device of claim 1, wherein said surface comprises a material selected from the group consisting of glass and plastic.

3. The device of claim 2, wherein said amplification primer immobilized to said surface is covalently immobilized to the surface of said solid support bead.

4. The device of claim 2, wherein said labeled hybridization probe immobilized to said surface is covalently immobilized to the surface of said solid support bead.

5. The device of claim 2, wherein said amplification primer and said labeled hybridization probe are each covalently immobilized to the surface of said solid support bead.

6. The device of claim 1, wherein said labeled hybridization probe comprises a fluorephore moiety and a quencher moiety.

7. The device of claim 1, wherein said device comprises two labeled hybridization probes immobilized to said surface, and wherein said two labeled hybridization probes comprise different sequences.

8. The device of claim 7, wherein prior to contact of said device with any nucleotide polymerizing enzyme there is immobilized to the surface of said solid support bead only one amplification primer sequence having a 3′

-end that can be extended by a DNA polymerase using as a template said first strand of the target nucleic acid or the complement thereof.

9. The device of claim 1, wherein prior to contact of said device with any nucleotide polymerizing enzyme there is immobilized to the surface of said solid support bead only one amplification primer sequence having a 3′

-end that can be extended by a DNA polymerase using as a template said first strand of the target nucleic acid or the complement thereof.

10. The device of claim 1, wherein said device is in contact with a solution comprising an RNA amplification product that is not immobilized to said device.