Chips for detecting DNA and method for detecting DNA using the same

• US 20040110214A1
• Filed: 10/03/2003
• Published: 06/10/2004
• Est. Priority Date: 12/10/2002
• Status: Abandoned Application

• Current Assignee: Electronics and Telecommunications Research Institute
• Original Assignee: Electronics and Telecommunications Research Institute
• Inventors: Kim, Sung Jin, Lee, Dae Sik, Park, Se Ho, Yang, Hae Sik, Kim, Yun Tae, Kim, Kyu Won, Shin, Yong Beom, Yoon, Tae Hwan
• Application Number:
  US10/677,315
• Publication Number:
  US 20040110214A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2537/1373 Displacement by a nucleic acid
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2565/525 characterised by the capture oligonucleotide being double stranded
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode

Abstract

The present invention provides a chip for detecting DNA and method for detecting DNA using the same. In the chip according to the present invention, a double strand formed by hybridization of signaling DNA and probe DNA is fixed to an electrode, wherein the signaling DNA and probe DNA are complementary each other and either of the two is complementary to sequence of a target DNA, and the complementarity between the target DNA, and the signaling DNA and the probe DNA is different, and electrochemically active compound is bonded to the signaling DNA. Further, the method for detecting DNA according to the present invention induces competitive reaction for performing hybridization of the probe DNA and the signaling DNA, and detects the target DNA by measuring change in an electrochemical signal of the signaling DNA. The chip for detecting DNA and the method for detecting the DNA according to the present invention have the same degree of sensitivity as a conventional detection method without direct marking on the target DNA, and have an advantage like that there exists no need for cleaning step in an electrochemical measurement step suitable for small-sized DNA detection system.

Claims

1. A chip for detecting DNA,

wherein a double strands formed by hybridization of signaling DNA and probe DNA is fixed to electrodes, and the signaling DNA and the probe DNA are complementary each other and either of the DNAs is also complementary to a base sequence of a target DNA, and the complementarities between the target DNA and the signaling DNA and the probe DNA are different, wherein the signaling DNA is labeled with an electrochemically active compound.

2. The chip for detecting DNA as claimed in claim 1,

wherein said electrode of the chip for detecting DNA is an array form having at least one electrode.

3. The chip for detecting DNA as claimed in claim 1,

wherein the chip for detecting DNA is made by fixing the probe DNA on the electrode by a method for forming a self-assembled monolayer, immersing said electrode to the signaling DNA solution, and deriving hybridization reaction by increasing temperature of the solution up to the melting point and decreasing it so that a double strand is formed.

4. The chip for detecting DNA as claimed in claim 1,

wherein the chip for detecting DNA is made by drawing hybridization reaction by increasing temperature of a solution containing same amounts of the probe DNA and the signaling DNA up to the melting point and cooling down gradually, so that a double strand is formed, and fixing the double strand on an electrode by a method for forming self-assembed monolayer.

5. The chip for detecting DNA as claimed in claim 1,

wherein complementarity between base sequences of the probe DNA and the target DNA is higher than that between base sequences of the probe DNA and the signaling DNA.

6. The chip for detecting DNA as claimed in claim 1,

wherein complementarity between base sequences of the signaling DNA and the target DNA is higher than that between base sequences of the probe DNA and the signaling DNA.

7. The chip for detecting DNA as claimed in claim 1,

wherein the electrically active material that is bonded to the signaling DNA is at least one selected from a group consisting of polypyridine or polyphenanthroline complex compound of ferrocene, rucenium, or osmium, viologen, hydroquinone, anthraquinone, and pyrrolo quinoline quinone.

8. The chip for detecting DNA as claimed in claim 1,

wherein the chip for detecting DNA is prepared by fixing at least two double strands that have been formed by using at least two probe DNAs and signaling DNAs corresponding to those probe DNAs to electrodes of electrode array having at least two electrodes arranged so that simultaneous multiple DNA detection can be performed.

9. A method for detecting DNA, comprising:

preparing a chip for detecting DNA, wherein a double strands formed by hybridization of signaling DNA and probe DNA is fixed to electrodes, and the signaling DNA and the probe DNA are complementary each other and either of the DNAs is also complementary to a base sequence of a target DNA, and the complementarities between the target DNA and the signaling DNA and the probe DNA are different, wherein the signaling DNA is labeled with an electrochemically active compound;

deriving competitive reaction among the probe DNA, the signaling DNA, and the target DNA by putting a solution of the target DNA to the chip for detecting DNA; and

detecting amount and type of the target DNA by measuring a change of the electrochemical signal of the signaling DNA before and after the competitive reaction.

10. The method for detecting DNA as claimed in claim 9,

in said step for deriving competitive reaction, heating electrode of the chip near to the melting temperature of a double strand formed by the signaling DNA and the probe DNA and cooling is repeated.

11. The method for detecting DNA as claimed in claim 9,

wherein the amount of the target DNA is quantified by measuring change of electrochemical signal before and after the competitive reaction of the signaling DNA.

12. The method for detecting DNA as claimed in claim 9,

wherein the type of the target DNA is determined by performing competitive reaction between a double strand and an unknown target DNA and measuring electrochemical signal produced from the competitive reaction, on a chip for DNA detection having electrode array of at least one electrode formed by fixing several double strands of the probe DNAs and the signaling DNAs corresponding to several types of expected base sequence of target DNAs.

13. The method for detecting DNA as claimed in claim 9,

wherein multiple DNA detection is performed by using the chip for detecting DNA that is formed by fixing at least two double strands of DNAs produced from at least two different probe DNAs and signaling DNAs corresponding to the probe DNAs to each electrode of an electrode array having at least two electrodes arranged.