DNA detection device

• US 6,861,216 B2
• Filed: 12/28/2000
• Issued: 03/01/2005
• Est. Priority Date: 12/28/1999
• Status: Expired due to Fees

• Current Assignee: Fujifilm Corporation (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Neriishi, Keiko, Umemoto, Chiyuki, Ogura, Nobuhiko, Hosoi, Yuichi
• Primary Examiner(s): Myers, Carla J.
• Application Number:
  US09/749,410
• Publication Number:
  US 20010026917A1
• Time in Patent Office: 1,524 Days
• Field of Search: 435 4- 6, 435/91.1, 435/91.2, 435/288, 382/100, 382/128, 382129-133, 382/181, 382/305, 382/312, 345/501, 345/502, 345/522, 345/507, 536 23- 29, 378/42, 378/190
• US Class Current: 435/6.12
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: G01T 1/2942 using autoradiographic methods

Abstract

A process for detecting a complementary DNA fragment is performed utilizing a DNA micro-array, a radiation image storage panel containing a stimulable phosphor, and a spacer sheet having plural openings.

Claims

1. A process for detecting a complementary DNA fragment which comprises the steps of:

bringing a liquid phase comprising single-stranded sample DNA fragments having a radioactive label into contact with a DNA micro-array having a support and at least two defined areas in each of which a group of probe compounds selected from the group consisting of DNA molecules, DNA fragments, synthesized oligonucleotides, synthesized polynucleotides, and PNA (peptide nucleic acid) are fixed under such condition that a group of the probe compounds fixed in one area differs from a group of the probe compounds fixed in another area, so that DNA fragments complementary to a group of the probe compounds are fixed to an area of the micro-array by hybridization of complementary DNA fragments to the probe compounds;

removing unfixed sample DNA fragments from the DNA micro-array;

keeping the DNA micro-array in contact with a radiation image storage panel containing a stimulable phosphor via a spacer sheet intervening between the DNA micro-array and the radiation image storage panel, said spacer sheet being in direct contact with the micro-array and having openings aligned with the ares of the micro-array to which the probe compounds are fixed, so that the radiation image storage panel can absorb and store radiation energy transmitted by the radioactive label of the fixed complementary DNA fragments through the openings in said spacer sheet;

irradiating the radiation image storage panel with a stimulating light, so that the image storage panel releases a stimulated emission from the area in which the radiation energy is stored;

detecting the stimulated emission photoelectrically to obtain a series of electric signals; and

processing the electric signals to locate the area in which the complementary DNA fragments are fixed.

2. The process of claim 1, in which the spacer sheet is made of non radiation-transmitting material.

3. The process of claim 1, in which the radiation image storage panel is irradiated with a stimulating light after it is separated from the DNA micro-array.

4. A process for detecting a complementary DNA fragment which comprises the steps of:

bringing a liquid phase comprising single-stranded sample DNA fragments having a radioactive lable into contact with a DNA micro-array having a support and at least two defined areas in each of which a group of probe compounds selected from the group consisting of DNA molecules, DNA fragments, synthesized oligonucleotides, synthesized polynucleotides, and PNA (peptide nucleic acid), are fixed under such conditions that a group of the probe compounds fixed in one area differs from a group of the probe compounds fixed in another area, so that DNA fragments complementary to a group of the probe compounds are fixed to an area of the micro-array by hybridization of complementary DNA fragments to the probe compounds;

removing unfixed sample DNA fragments from the DNA micro-array;

keeping the DNA micro-array in contact with a radiation image storage panel containing a stimulable phosphor via a spacer sheet intervening between the DNA micro-array and the radiation image storage panel, said spacer sheet being in direct contact with the micro-array and having openings aligned with the areas of the micro-array to which the probe compounds are fixed, so that the radiation image storage panel can absorb and store radiation energy transmitted by the radioactive label of the fixed complementary DNA fragments through the openings in said spacer sheet;

irradiating the radiation image storage panel with a stimulating light, so that the image storage panel releases a stimulated emission from the area in which the radiation energy is stored;

detecting the stimulated emission photoelectrically to obtain a series of electric signals; and

processing the electric signals to locate the area in which the complementary DNA fragments are fixed,

wherein said spacer sheet has a thickness in the range of 10 to 300 μ

m and is made of a non radiation-transmitting material selected from the group consisting of aluminum, brass, stainless steel, polyethylene terephthalate and polyethylene naphthalate.