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

This application is a division of U.S. patent application Ser. No. 11/414,022, filed Apr. 28, 2006, which claims priority to Korean Patent Application No. 10-2005-0055904, filed on Jun. 27, 2005, the disclosure of each of which is incorporated by reference herein in its entirety.

1. Field of the Invention

The present invention relates to a method for highly sensitive nucleic acid detection using a nanopore and a non-specific nucleic acid-binding agent.

2. Description of the Related Art

Various methods of detecting a target biomolecule in a sample have been reported. A method using nanopores is widely used in a highly sensitive DNA detecting system, which is an imitation of a bio-pore system, and is capable of sequencing bases in nucleic acids.

A method of characterizing individual polymer molecules based on monomer-interface interactions is disclosed in U.S. Pat. No. 6,362,002. In this method, a single-stranded nucleic acid polymer can pass through a channel in an interface between two pools, but a double-stranded nucleic acid polymer cannot pass through the channel, and can thus be detected.

A method of determining the presence of double-stranded nucleic acids in a sample is disclosed in U.S. Pat. No. 6,428,959. Double-stranded nucleic acid can be detected by translocating nucleic acids in a sample through a nanopore, monitoring a current amplitude through the nanopore during the translocation, and measuring the duration of a transient blockade of current. However, U.S. Pat. No. 6,428,959 does not disclose a method of detecting nucleic acids using a non-specific nucleic acid-binding agent to increase a current amplitude.

A method for the characterization of nucleic acid molecules is disclosed in U.S. Patent Publication No. 2003/0104428. A maximum change in a signal can be achieved by changing specific local areas using proteins specific to nucleotide sequences. However, U.S. Patent publication No. 2003/0104428 relates to the detecting of DNA having a specific sequence using nanopore, and does not disclose the detecting of nucleic acids using a non-specific nucleic acid-binding agent to increase a current amplitude.

The inventors of the present invention discovered that a nucleic acid can be accurately detected by increasing a current amplitude change through nanopores using a non-specific nucleic acid binding agent, regardless of specific sequences.

The present invention provides a method of sensitively detecting nucleic acids in a nucleic acid sample. The method includes: contacting the sample including the nucleic acid with a non-specific nucleic acid binding agent in an electrically conductive fluid medium; contacting the sample including 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 present invention also provides an apparatus for detecting nucleic acids. The apparatus includes: 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.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and other features and advantages of the present invention will become more apparent by describing in detail exemplary embodiments thereof with reference to the attached drawings in which:

FIG. 1 illustrates a measured current through a nanopore using only an intercalator without lambda DNA;

FIG. 2 illustrates a measured current through a nanopore A in Example 2 using the intercalator with lambda DNA;

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