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Electrochemical detection method of complementarity to nucleic acid bases

Foreign code F110005204
File No. A161-03US
Posted date Aug 25, 2011
Country United States of America
Application number 47165701
Gazette No. 20050153285
Gazette No. 7169614
Date of filing Mar 13, 2001
Gazette Date Jul 14, 2005
Gazette Date Jan 30, 2007
International application number JP2001001974
International publication number WO2002073183
Date of international filing Mar 13, 2001
Date of international publication Sep 19, 2002
Priority data
  • 2001WO-JP01974 (Mar 13, 2001) WO
Title Electrochemical detection method of complementarity to nucleic acid bases
Abstract (US7169614)
A convenient and commonly applicable method for the specific detection of a nucleic acid with an arbitrary sequence is provide.
This method comprises attaching at least a nucleic acid single strand to an electrode, bringing the thus-obtained modified electrode in contact with a solution containing the analyte single-stranded nucleic acid, and measuring the redox reaction of the redox marker.
Scope of claims [claim1]
1. A method for electrochemical detection of nucleobase pair complementarity, which comprises: contacting a modified electrode obtained by bonding at least a single stranded nucleic acid to an electrode so that the single-stranded nucleic acid is oriented vertically with respect to the surface of the electrode and the electrode is a gold electrode, with a solution containing an analyte single stranded nucleic acid and a redox marker, wherein the redox marker is [Mo (CN)6]4/13- or [Fe (CN)6]4/13-;
and
detecting the disappearance of an electric signal generated by reaction between the modified electrode and the redox marker, when the single stranded nucleic acid on the modified electrode and the analyte single stranded nucleic acid are complementary.
[claim2]
2. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the electrode is a gold disk electrode.
[claim3]
3. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein a molecule that is not complementary to the analyte single stranded nucleic acid is bonded to the modified electrode along with the single stranded nucleic acid for vertically orienting the single stranded nucleic acid with respect to the surface of the electrode.
[claim4]
4. The method for electrochemical detection of nucleobase pair complementarity of claim 3, wherein the single stranded nucleic acid and the molecule that is not complementary to the analyte single stranded nucleic acid form a mixed monolayer on the surface of the electrode.
[claim5]
5. The method for electrochemical detection of nucleobase pair complementarity of claim 3, wherein the molecule that is not complementary to the analyte single stranded nucleic acid is 6-mercapto-1-hexanol.
[claim6]
6. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the single stranded nucleic acid is a single stranded peptide nucleic acid.
[claim7]
7. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the concentration of the analyte single stranded nucleic acid is 10-11 to 10-2 M.
[claim8]
8. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the analyte single stranded nucleic acid is a single stranded DNA.
[claim9]
9. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the redox marker is [Fe(CN)6]4-/3-.
[claim10]
10. The method for electrochemical detection of nucleobase pair complementarity of claim 9, wherein the concentration of [Fe(CN)6]4-/3- is 0.1 to 1 mM.
[claim11]
11. The method for electrochemical detection of nucleobase pair complementarity of claim 1, wherein the redox reaction is measured by voltammetry.
[claim12]
12. A modified electrode for detecting nucleobase pair complementarity by the method of claim 1, which comprises a mixed monolayer of a peptide nucleic acid and a molecule that is not complementary to the analyte nucleic acid, formed on the surface of a gold disk electrode.
  • Inventor, and Inventor/Applicant
  • UMEZAWA YOSHIO
  • AOKI HIROSHI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Endocrine Disrupters AREA
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