Nucleic-acid probe substrate, system for temperature control of the substrate, and gene detection method making use of the same

• US 20040009525A1
• Filed: 06/23/2003
• Published: 01/15/2004
• Est. Priority Date: 06/24/2002
• Status: Active Grant

• Current Assignee: Canon Ayutthaya Limited (Canon Inc.)
• Original Assignee: Canon Ayutthaya Limited (Canon Inc.)
• Inventors: Kawaguchi, Masahiro
• Granted Patent: US 7,713,692 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/00376 Conductive heating, e.g. heated plates

Abstract

A system used to detect and assay genes by using a nucleic-acid probe substrate is disclosed, which can be used in highly general-purpose genetic chips usable for various purposes, and has a temperature control means enabling control of temperature of the whole genetic chip in a high reproducibility and uniformity, and further even satisfies low-cost performance. A method for detecting and assaying gene nucleic acids by using such a system is also disclosed. A heat-conductive material made up using a material having good thermal conductivity, having a shape having general-purpose properties, is used as the temperature control means, and is disposed in such a form that it covers substantially the whole substrate face or stands in contact therewith, on the back of the substrate, or on the side facing the surface to which the detecting single-stranded nucleic acid fragments have been immobilized. This makes it possible to improve the diffusion of heat in the in-plane direction of the whole substrate face. In addition, heat is given and received through such a heat-conductive material.

Claims

1. A system for temperature control of a nucleic-acid probe substrate, which controls the temperature to the substrate surface of which a plurality of nucleic-acid probes containing single-stranded nucleic acid fragments having a complementary sequence in respect to a target DNA have been immobilized in order that the target DNA contained in a specimen is detected according to hybridization;

the system comprising;

a heat conduction means comprising a heat-conductive material disposed on the back of the substrate to the substrate surface of which the plurality of single-stranded nucleic acid fragments have been immobilized, and in contact with the back of the substrate;

a heating means or cooling means which is provided in contact with the heat-conductive material; and

a means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material, to control the temperature of the heat-conductive material;

the temperature of the substrate disposed in contact being controlled through the temperature control of the heat-conductive material.

3. The system according to claim 1 or 2, wherein said heat-conductive material is formed of any one of a metal and a resin or a composite of these two or more.

2. A system for temperature control of a nucleic-acid probe substrate, which controls the temperature of a substrate to the substrate surface of which a plurality of nucleic-acid probes containing single-stranded nucleic acid fragments having a complementary sequence in respect to a target DNA have been immobilized in order that the target DNA contained in a specimen is detected according to hybridization;

the system comprising;

a heat conduction means comprising a heat-conductive material disposed on the surface of the substrate to the substrate surface of which the plurality of single-stranded nucleic acid fragments have been immobilized, facing, and in contact with, the substrate surface, partly leaving a space for feeding the specimen thereinto;

a heating means or cooling means which is provided in contact with the heat-conductive material; and

a means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material to control the temperature of the heat-conductive material;

the specimen fed into the space and the substrate surface, which are in contact with the heat-conductive material, being temperature-controlled through the temperature control of the heat-conductive material.

4. A method for detecting genes by utilizing as a detection means a substrate to the substrate surface of which a plurality of nucleic-acid probes containing single-stranded nucleic acid fragments having a complementary sequence in respect to a target DNA have been immobilized in order that the target DNA contained in a specimen is detected according to hybridization;

the method comprising;

disposing a heat-conductive material on the back of the substrate to the substrate surface of which the plurality of single-stranded nucleic acid fragments have been immobilized, and in contact with the back of the substrate;

disposing a heating means or cooling means in contact with the heat-conductive material; and

providing a temperature control means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material to control the temperature of the heat-conductive material;

the detection being operated while the substrate standing bonded sandwichedly and the specimen standing in contact with the substrate surface are temperature-controlled through the temperature control of the heat-conductive material by the temperature control means during the operation of gene detection.

5. The method according to claim 4, wherein, in a plurality of steps involved in the gene detection operation, said substrate and said specimen standing in contact with the substrate surface are temperature-controlled;

and

the temperature in the plurality of steps requiring temperature control is successively controlled by the temperature control means which utilizes said heating means.

6. The method according to claim 4, wherein, in a plurality of steps involved in the gene detection operation, said substrate and said specimen standing in contact with the substrate surface are temperature-controlled;

and

the temperature in the plurality of steps requiring temperature control is successively controlled by the temperature control means which utilizes said cooling means.

7. The method according to claim 4, wherein, as said heat-conductive material, which is utilized for the temperature control the substrate and of the specimen standing in contact with the substrate surface, a heat-conductive material is used which is formed of any one of a metal and a resin or a composite of these two or more.

8. A method for detecting genes by utilizing as a detection means a substrate to the substrate surface of which a plurality of nucleic-acid probes containing single-stranded nucleic acid fragments having a complementary sequence in respect to a target DNA have been immobilized in order that the target DNA contained in a specimen is detected according to hybridization;

the method comprising;

disposing a heat-conductive material on the surface of the substrate to the substrate surface of which the plurality of single-stranded nucleic acid fragments have been immobilized, facing, and in contact with, the substrate surface, partly leaving a space for feeding the specimen thereinto;

disposing a heating means or cooling means in contact with the heat-conductive material; and

providing a temperature control means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material to control the temperature of the heat-conductive material;

the detection being operated while the specimen fed into the space and the substrate surface, which are in contact with the heat-conductive material, being temperature-controlled through the temperature control of the heat-conductive material by the temperature control means during the operation of gene detection.

9. The method according to claim 8, wherein, in a plurality of steps involved in the gene detection operation, said substrate and said specimen standing in contact with the substrate surface are temperature-controlled;

and

the temperature in the plurality of steps requiring temperature control is successively controlled by the temperature control means which utilizes said heating means.

10. The method according to claim 8, wherein, in a plurality of steps involved in the gene detection operation, said substrate and said specimen standing in contact with the substrate surface are temperature-controlled;

and

the temperature in the plurality of steps requiring temperature control is successively controlled by the temperature control means which utilizes said cooling means.

11. The method according to claim 8, wherein, as said heat-conductive material, which is utilized for the temperature control of the substrate and the specimen standing in contact with the substrate surface, a heat-conductive material is used which is formed of any one of a metal and a resin or a composite of these two or more.

12. A probe substrate temperature control system for controlling the temperature of a probe substrate to the substrate surface of which a plurality of probes bindable specifically to a target substance have been immobilized in order to detect the target substance;

the system comprising;

a heat conduction means comprising a heat-conductive material disposed on the side opposite to the surface of the probe substrate to which surface the detecting target substance have been immobilized, and in contact with the back of the substrate;

a heating means or cooling means which is provided in contact with the heat-conductive material; and

a means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material to control the temperature of the heat-conductive material;

the temperature of the substrate disposed in contact being controlled through the temperature control of the heat-conductive material.

13. A probe substrate comprising:

a substrate;

a plurality of probes bindable specifically to a target substance which have been immobilized to the substrate surface; and

a heat-conductive material for controlling the temperature of the substrate;

the material being disposed in contact with the back of the substrate.

14. A probe substrate temperature control system for controlling the temperature of a probe substrate to the substrate surface of which a plurality of probes bindable specifically to a target substance have been immobilized in order to detect the target substance;

the system comprising;

a heat conduction means comprising a heat-conductive material disposed on the surface of the substrate to the substrate surface of which the plurality of probes have been immobilized, facing, and in contact with, the substrate surface, partly leaving a space for feeding the specimen thereinto;

a heating means or cooling means which is provided in contact with the heat-conductive material; and

a means for controlling the amount of heat flowing across the heating means or cooling means and the heat-conductive material to control the temperature of the heat-conductive material;

the temperature of the substrate disposed in contact being controlled through the temperature control of the heat-conductive material.

15. A probe substrate comprising:

a substrate;

a plurality of probes bindable specifically to a target substance which have been immobilized to the substrate surface; and

a heat-conductive material for controlling the temperature of the substrate;

the material being disposed on the surface of the substrate to the substrate surface of which the plurality of probes have been immobilized, facing, and in contact with, the substrate surface, partly leaving a space for feeding the specimen thereinto.