Detection of a target in a sample

• US 20030203394A1
• Filed: 06/03/2003
• Published: 10/30/2003
• Est. Priority Date: 05/04/1998
• Status: Abandoned Application

• Current Assignee: Erez Braun, Uri Sivan, Yoav Eichen
• Original Assignee: Erez Braun, Uri Sivan, Yoav Eichen
• Inventors: Sivan, Uri, Braun, Erez, Eichen, Yoav
• Application Number:
  US10/452,139
• Publication Number:
  US 20030203394A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

The invention concerns a system, device, kit and method for detecting the presence, or concentration of a target in a sample. An assay set comprising at least two spaced apart electrodes is used, comprising a recognition moiety, capable of specific binding to the target, which is attached to at least one of the electrodes or the substrate therein between. If the recognition moiety binds the target then a conductive bridge can be formed between the electrodes, based on the complex between the recognition moiety and the target. The conductive bridge is formed by using nucleation-center forming entities attached to said complexes or to said targets from which a conductive substance is substantially grown. Alternatively the conducting bridge forms a conductive polymer between the electrodes.

Claims

1. A method for detecting a target nucleic acid molecule in a sample, said method comprising:

providing a device for detecting the presence of a target nucleic acid molecule in a sample, comprising;

two electrical conductors, including a first electrical conductor and a second electrical conductor, but where the electrical conductors are not in contact, and

one or more sets of two oligonucleotide probes attached to the electrical conductors, where the oligonucleotide probes are positioned such that the probes cannot come into contact with one another and such that a target nucleic acid molecule, which has two sequences, a first sequence complementary to a first probe attached to the first electrical conductor and a second sequence complementary to a second probe attached to the second electrical conductor, can bind to both probes concurrently;

contacting the probes with a sample which may have the target nucleic acid molecule under selective hybridization conditions to permit target nucleic acid molecules, if any, present in the sample to hybridize to both of the probes;

forming a conductive coating over the oligonucleotide probes and any target nucleic acid molecules hybridized to the oligonucleotide probes; and

determining if an electrical current can be carried between the probes, said electrical current between the probes indicating the presence of the target nucleic acid molecule in the sample which has sequences complementary to the probes.

2. The method according to claim 1, wherein the target nucleic acid molecule is DNA.

3. The method according to claim 1, wherein the target nucleic acid molecule is RNA.

4. The method according to claim 1, wherein the conductor is silver.

5. The method according to claim 1, wherein the conductor is gold.

6. The method according to claim 1, wherein the probes are complementary to sequences from the genetic material of a pathogenic bacteria.

7. A method for detecting a target nucleic acid molecule in a sample, said method comprising:

providing a sample potentially containing a target nucleic acid molecule;

providing a pair of oligonucleotide probes which are spaced apart by a gap, wherein the oligonucleotide probes are capable of hybridizing to spaced apart portions of the target nucleic acid molecule;

contacting the sample with the spaced apart pair of oligonucleotide probes under conditions effective to permit any of the target nucleic acid molecule in the sample to hybridize to both of the spaced apart oligonucleotide probes, thereby bridging the gap and coupling the pair of oligonucleotide probes with the hybridized target nucleic acid molecule, if any;

forming a conductive coating over the pair of oligonucleotide probes and any hybridized target nucleic acid molecule; and

determining whether electrical current passes between the pair of oligonucleotide probes as a result of said contacting, thereby indicating a presence of the target nucleic acid molecule in the sample.

8. A method according to claim 7, wherein the conductor is selected from the group consisting of gold and silver.

9. A method according to claim 7, wherein a plurality of each pair of oligonucleotide probes is provided, said method further comprising:

identifying the number of pairs of identical oligonucleotide probes between which electrical current passes to determine the concentration of the target nucleic acid molecule present in the sample.

10. A method according to claim 7, wherein the pair of oligonucleotide probes are immobilized on a substrate.

11. A method according to claim 10, wherein the substrate is selected from the group consisting of glass, silicon, and polymeric material.

12. A method according to claim 10, wherein the pair of oligonucleotide probes are attached to the substrate with a linker.

13. A method according to claim 7, wherein the target nucleic acid molecule is selected from the group consisting of DNA, RNA, and chemically modified nucleic acid molecules.

14. A method according to claim 7, wherein the sample is blood.

15. A method according to claim 7, wherein said method is used to detect infectious agents.