Method for highly sensitive nucleic acid detection using nanopore and non-specific nucleic acid-binding agent

• US 20060292605A1
• Filed: 04/28/2006
• Published: 12/28/2006
• Est. Priority Date: 06/27/2005
• Status: Active Grant

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Min, Jun-hong, Lee, In-ho, Kim, Kui-hyun, Kim, Ah-gi
• Granted Patent: US 7,504,261 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Provided is a method of sensitively detecting nucleic acids in a nucleic acid sample, the method comprising: contacting the sample comprising the nucleic acid with a non-specific nucleic acid binding agent in an electrically conductive fluid medium; contacting the sample comprising the nucleic acid bound to the agent with a nanopore; and applying a voltage to the nanopore and monitoring a current change through the nanopore. The nucleic acid can be sensitively detected because a change in current amplitude through the nanopore is greater than when nucleic aid detection is performed without using an intercalator.

Claims

1. A method of sensitively detecting nucleic acids in a nucleic acid sample, the method comprising:

contacting the sample comprising the nucleic acid with a non-specific nucleic acid binding agent in an electrically conductive fluid medium;

contacting the sample comprising the nucleic acid bound to the agent with a nanopore; and

applying a voltage across the nanopore and monitoring a current change through the nanopore.

2. The method of claim 1, wherein the non-specific nucleic acid binding agent is selected from the group consisting of an intercalator, a major groove binding agent and a minor groove binding agent.

3. The method of claim 2, wherein the intercalator is selected from the group consisting of YOYO (1,1′

-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzoxazolylidene)methyl]]-,tetraiodide), TOTO (1-1′

-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzothiazolylidene)methyl]]-,tetraiodide) and Ethidium Bromide (EtBr).

4. The method of claim 1, wherein the nanopore has a diameter ranging from 1.5 nm to 120 nm.

5. The method of claim 1, wherein the voltage ranges from 100 mV to 1000 mV.

6. The method of claim 1, wherein the electrically conductive fluid medium comprises multiple nucleic acids comprising different sequences.

7. An apparatus for detecting nucleic acids, the apparatus comprising:

a nanopore;

a non-specific nucleic acid binding agent;

a device applying a voltage across the nanopore; and

a detector monitoring a current change through the nanopore.

8. The apparatus of claim 7, wherein the non-specific nucleic acid binding agent is selected from the group consisting of an intercalator, a major groove binding agent and a minor groove binding agent.

9. The apparatus of claim 8, wherein the intercalator is selected from the group consisting of YOYO (1,1′

-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzoxazolylidene)methyl]]-,tetraiodide), TOTO (1-1′

-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzothiazolylidene)methyl]]-,tetraiodide) and Ethidium Bromide (EtBr).

10. The apparatus of claim 7, wherein the nanopore has a diameter ranging from 1.5 nm to 120 nm.

11. The apparatus of claim 7, wherein the voltage ranges from 100 mV to 1000 mV.