Specific base sequence detection method and primer extension reaction detection method

• US 20060105366A1
• Filed: 10/13/2005
• Published: 05/18/2006
• Est. Priority Date: 11/16/2004
• Status: Abandoned Application

• Current Assignee: Seiko Epson Corporation (Seiko Group)
• Original Assignee: Seiko Epson Corporation (Seiko Group)
• Inventors: Yokokawa, Shinobu, Fukushima, Hitoshi, Takiguchi, Hiroshi, Hiroshima, Shinichi
• Application Number:
  US11/248,241
• Publication Number:
  US 20060105366A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode

Abstract

A specific base sequence detection method, comprising preparing a sample solution including a target nucleic acid, a primer for amplifying a specific base sequence and whose end has a site to be coupled to an electrode, and nucleotide; extending the primer if the specific base sequence is present in the nucleic acid by putting the sample solution in a condition that causes an extension reaction of the primer; performing an electrical measurement by immersing an electrode in a measurement solution including the sample solution that has completed the extension reaction; and detecting whether the specific base sequence is present in the nucleic acid based on a result of the electrical measurement.

Claims

1. A specific base sequence detection method, comprising:

preparing a sample solution including a target nucleic acid, a primer for amplifying a specific base sequence and whose end has a site to be coupled to an electrode, and nucleotide;

extending the primer if the specific base sequence is present in the nucleic acid by putting the sample solution in a condition that causes an extension reaction of the primer;

performing an electrical measurement by immersing an electrode in a measurement solution including the sample solution that has completed the extension reaction; and

detecting whether the specific base sequence is present in the nucleic acid based on a result of the electrical measurement.

2. The specific base sequence detection method according to claim 1, the primer being composed of a complementary sequence that binds to the specific base sequence in a complementary manner.

3. The specific base sequence detection method according to claim 1, a result of detecting whether the primer has been extended based on a result of the electrical measurement being used to detect whether the specific base sequence is present.

4. The specific base sequence detection method according to claim 1, the primer including an upstream primer and a downstream primer at least one of whose end has the site to be coupled to an electrode.

5. The specific base sequence detection method according to claim 1, the site to be coupled to an electrode being one of a thiol group, an amino group, and biotin.

6. The specific base sequence detection method according to claim 1, the electrical measurement being one of a measurement of impedance, current, and electrical charge of the electrode.

7. A primer extension reaction detection method, comprising:

preparing a sample solution including a target nucleic acid, a primer for amplifying a specific base sequence and whose end has a site to be coupled to an electrode, and nucleotide;

extending the primer if the specific base sequence is present in the nucleic acid by putting the sample solution in a condition that causes an extension reaction of the primer;

performing an electrical measurement by immersing an electrode in a measurement solution that includes the sample solution that has completed the extension reaction; and

detecting whether the primer has been extended based on a result of the electrical measurement.