Gene detection method

• US 5,972,692 A
• Filed: 06/30/1997
• Issued: 10/26/1999
• Est. Priority Date: 09/28/1990
• Status: Expired due to Fees

• Current Assignee: Kabushiki Kaisha Toshiba (Toshiba Corporation)
• Original Assignee: Kabushiki Kaisha Toshiba (Toshiba Corporation)
• Inventors: Ishimori, Yoshio, Ito, Keiko, Hashimoto, Koji
• Primary Examiner(s): Campbell, Eggerton A.
• Application Number:
  US08/886,161
• Time in Patent Office: 848 Days
• Field of Search: 435/3, 435/287.2, 435/285.2
• US Class Current: 435/285.2
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C12Q 2565/519 characterised by the capture moiety being a single stranded oligonucleotide
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 27/3276 being a hybridisation with immobilised receptors using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68

Abstract

A single stranded nucleic acid probe having a base sequence complementary to the gene to be detected is immobilized onto the surface of an electrode or the tip of an optical fiber, and the nucleic probe is reacted with the gene sample denatured to a single stranded form, and then the nucleic acid probe hybridized with the gene is detected. In this procedure, to the reaction system consisting of the nucleic acid probe and the gene sample, a double stranded nucleic acid recognizing substance capable of binding specifically to the double stranded nucleic acid and being active electrochemically or optically is added. The detection of the nucleic acid probe is conducted by electrochemical or optical determination utilizing the electrode or optical fiber mentioned above. By this method, safer and more convenient detection of the gene is possible at a higher sensitivity even in a reduced time period.

Claims

1. A gene-detection device comprising:

an electrode having a single stranded nucleic acid probe immobilized on its surface which has a base sequence complementary to the gene to be detected;

a reaction vessel in which a sample solution containing a gene sample which has been denatured into a single stranded form can react with said electrode to form a double stranded nucleic acid on the surface of the electrode;

washing means to remove unreacted gene sample by washing the gene sensor after hybridization of the electrode with the gene sample; and

a detection vessel to store a substance which binds specifically to the double stranded nucleic acid formed on the surface of the electrode and which is active physiochemically to the reaction system of said nucleic acid probe in said gene sample.

2. The gene-detection device of claim 1, further comprising a gene sample purification means for preparing the sample solution, wherein the gene sample is denatured into a single-stranded form.

3. The gene-detection device of claim 1, wherein said reaction vessel is divided into a purality of vessels, each of which contains a different sample solution, thereby enabling a purality of samples to be analyzed simultaneously.

4. The gene-detection device of claim 1, further comprising transportation means to transport the electrode, and means to control the temperature of the sample solution.

5. The gene-detection device of claim 4, further comprising a gene sample purification means for preparing the sample solution, wherein the gene sample is denatured into a single-stranded form.

6. The gene-detection device of claim 4, wherein said reaction vessel is divided into a purality of vessels, each of which contains a different sample solution, thereby enabling a purality of samples to be analyzed simultaneously.

7. A device according to claim 1, further comprising a dissociation means to dissociate the double stranded nucleic acid formed on the surface of the gene sensor into the nucleic acid probe immobilized on the surface of the gene sensor and the single stranded gene sample and to remove the gene sample, whereby regenerating the gene sensor.