Gene examining apparatus and method of detecting target nucleic acid using the same

• US 20070134712A1
• Filed: 01/19/2007
• Published: 06/14/2007
• Est. Priority Date: 07/31/2001
• Status: Abandoned Application

• Current Assignee: Olympus Corporation
• Original Assignee: Olympus Corporation
• Inventors: Sakamoto, Hiroko, Shibazaki, Takami, Yonekawa, Hiroyuki, Satoh, Takatomo
• Application Number:
  US11/655,316
• Publication Number:
  US 20070134712A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

The present invention relates to a gene examining apparatus utilizing a computer, the apparatus comprising (1) DNA microarrays in each of which a large number of fine liquid accommodating sections are two-dimensionally arranged so that openings of the fine liquid accommodating sections are located on the same plane, in which each of the liquid accommodating sections can three-dimensionally accommodate a liquid, and in which hybridization reaction occurs in the liquid accommodating section between a target nucleic acid already labeled with an optical marker substance and the nucleic acid probe, and (2) a microscope comprising a stage supporting the DNA microarrays set forth in (1), a temperature regulating section that regulates the temperature of each DNA microarray, and imaging means for picking up an image of an optical signal from the DNA microarray.

Claims

1-22. -22. (canceled)

23. A method of examining a gene using an apparatus including;

a DNA microarray including;

a plurality of liquid accommodating sections, and

a porous film to contact with said liquid in the liquid accommodating section, wherein openings of said liquid accommodating sections are located on the same plane and said porous film has channels;

a microscope to observe the plane where said openings are located;

a fluid transporting section that moves the fluid into and out of said DNA microarray; and

a computer that controls picking up an image with said microscope and moving the fluid with said fluid transporting section, the method comprising;

a step of providing said DNA microarray with a fluid which includes a nucleic acid added with an optical marker substance; and

a step of withdrawing the fluid from the DNA microarray through said fluid transporting section after the step of providing, but before the step of picking up the image is performed,

a step of picking up the image of the DNA microarray with said microscope.

24. The method of examining a gene using the apparatus according to claim 23, wherein the DNA microarray further including a nucleic acid probe that is immobilized on an inner wall of each of said channels.

25. The method of examining a gene using the apparatus according to claim 23, the method comprising;

(1) amplifying a nucleic acid extracted from a tissue or a cell obtained from a subject and adding an optical marker substance as a marker substance to afford a labeled nucleic acid;

(2) adding the labeled nucleic acid obtained in (1) to a DNA microarray comprising a desired nucleic acid probe;

(3) using the DNA microarray in (2) to cause hybridization reaction under desired conditions;

(4) after the reaction in (3) has been finished, collecting a solution contained in the DNA microarray, at the bottom of the array;

(5) measuring the intensity of an optical signal from the optical marker substance in the DNA microarray obtained in (3);

(6) agitating again the liquid collected at the bottom of the array in (4);

(7) using the DNA microarray in (6) to repeat hybridization reaction under desired conditions as required;

(8) measuring the intensity of an optical signal from the DNA microarray obtained in (6); and

(9) on the basis of the intensities of the optical signals obtained in (4) and (8), determining the amount of an expressed gene and/or whether or not any mutated gene is present, to obtain results of gene examinations.

26. The method according to claim 25, wherein operations from (5) to (8) are repeated twice or more.

27. The method of examining a gene using the apparatus according to claim 23 or 25 the method further comprising a step of pumping the fluid into and out of said DNA microarray.