Microelectronic device for electrochemical detection of nucleic acid hybridization

• US 7,049,068 B2
• Filed: 11/06/2001
• Issued: 05/23/2006
• Est. Priority Date: 06/27/1995
• Status: Expired due to Fees

• Current Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Original Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Inventors: Thorp, H. Holden, Johnston, Dean H., Napier, Mary E., Loomis, Carson R., Sistare, Mark F., Kim, Jinheung
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US10/008,233
• Publication Number:
  US 20020106683A1
• Time in Patent Office: 1,659 Days
• Field of Search: 435/6, 435/91.2, 435/320.1, 435/174, 435/283.1, 435/287.2, 530/350, 536/23.4, 536/23.1, 536/24.3, 422/68.1, 422/82.01
• US Class Current: 435/6.13
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2525/117 incorporating modified base
• CPC Class Codes: C12Q 2525/197 incorporating a spacer/coupling moiety
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

A method of detecting a nucleic acid (e.g., DNA, RNA) that contains at least one preselected base (e.g., adenine, guanine, 6-mercaptoguanine, 8-oxo-guanine, and 8-oxo-adenine) comprises (a) reacting the nucleic acid with a transition metal complex capable of oxidizing the preselected base in an oxidation-reduction reaction; (b) detecting the oxidation-reduction reaction; and (c) determining the presence or absence of the nucleic acid from the detected oxidation-reduction reaction at the preselected base. The method may be used in a variety of applications, including DNA sequencing, diagnostic assays, and quantitative analysis.

Claims

1. A microelectronic device useful for the electrochemical detection of a nucleic acid species, said device comprising:

a microelectronic substrate having first and second opposing faces;

a plurality of conductive oxidation-reduction detection electrodes on said first face; and

a plurality of oligonucleotide capture probes immobilized on a non-conductive layer on said first face adjacent said oxidation-reduction electrodes;

with each of said plurality of oligonucleotide capture probes positioned adjacent a different oxidation-reduction electrode;

and with each of said plurality of oligonucleotide capture probes and said oxidation-reduction detection electrodes electrically connected by an aqueous solution, said aqueous solution having a transition metal complex therein;

and wherein said aqueous solution connecting each of said plurality of oligonucleotide capture probes and said oxidation-reduction detection electrodes is the same.

2. A microelectronic device according to claim 1, further comprising a contact electrically connected to said oxidation-reduction electrode.

3. A microelectronic device according to claim 1, wherein said substrate is silicon.

4. A microelectronic device according to claim 1, wherein said oligonucleotide capture probe is from 4 to 100 nucleotides in length.