WO2002004951A1

WO2002004951A1 - Micro-array

Info

Publication number: WO2002004951A1
Application number: PCT/JP2001/002821
Authority: WO
Inventors: Akio Yamane; Toru Miyagi
Original assignee: Wakunaga Pharmaceutical Co., Ltd.
Priority date: 2000-07-10
Filing date: 2001-03-30
Publication date: 2002-01-17
Also published as: AU2001244687A1

Abstract

A micro-array excellent in operability and data reliability, and suitable for mass specimen-treating. A micro-array for detecting a target molecule, comprising at least one unit consisting of a carrier for detecting a target molecule and allowing a target-molecule-capturable molecule (capturing molecule) to be fixed, and a frame for retaining a sample on the carrier.

Description

Description Microarray Background of the Invention

Field of the invention

The present invention relates to a microarray used for analyzing biomolecules, and more particularly, to a diagnostic microarray suitable for processing a large amount of a sample.

Related technology

In recent years, breakthroughs in DNA sequencing technology have revealed a large number of gene sequences. However, in many cases, it is difficult to specify the function of a gene using only the nucleotide sequence, so expression analysis must be performed.

Analysis of gene expression at the mRNA level can be performed by analyzing a specific known gene sequence for a sample, as in the Northern blot method (Alwine JC et al. Proc Natl Acad Sci USA. 1977 Dec; 74 (12): 5350-4). There is a method used as a probe. On the other hand, conversely, genes with different expression levels among different samples, such as the differential display method (Liang P and Pardee AB. Science. 1992 Aug 14; 257 (5072): 967-71), are used. There are ways to find them.

Northern blotting is time-consuming and labor-intensive, and the number of genes that can be analyzed is limited. Although it is a reliable and convenient method for analyzing a small number of candidate genes, it is very difficult to analyze the vast number of genes identified one after another. On the other hand, in the differential display method, a sample extracted from cells and tissues in different states and a sample amplified at random are electrophoresed, and band patterns are compared to extract bands with different intensities between different samples. And determine the base sequence. In many cases, this band does not contain the entire nucleotide sequence of the gene, and the gene is identified by comparing the partial sequence obtained from the band with the previously determined gene sequence data. You. The function can be estimated from the state of the identified gene that was strongly expressed in the cell.

However, if the sequence obtained is not known, clone from the library. There must be. If there is a known sequence, the gene can be identified quickly, but the number of genes that can be monitored for changes in the expression level is limited due to the limitations of electrophoretic resolution.

A technique called DNA microarray or DNA chip is capable of analyzing a much larger number of genes than these methods.

A DNA microarray is a solid plate in which a partial amplification product of a known gene sequence or an oligonucleotide is immobilized on a small plate such as a slide glass at a high density. Thousands to tens of thousands of types are obtained by hybridization. This technology allows simultaneous detection of gene expression levels on a single slide. DNA can be directly solid-phase synthesized on a carrier, or can be immobilized on a carrier by spotting a previously synthesized DNA with a fine pin or the like.

DNA microarrays are being developed as a very powerful technique in expression analysis and polymorphism analysis research, and clinical applications are expected to become more active in the future based on these research results ( Hodgson J. Nat Biotechnol. 1998 Aug; 16 (8): 725-7).

However, in the hybridization of a conventional DNA microarray, as described in Schena M et al. Science. 1995 Oct 20; 270 (5235): 467-70, the probe is placed on a slide glass. In general, the sample solution is spread over the entire probe area by immobilizing a solid phase, placing a hybridization solution containing a detection target on a probe, and covering the probe with a cover slip. The target nucleic acid group is labeled with a fluorescent substance or the like, and after hybridization, excess labeled nucleic acid not hybridized to the probe is removed by washing.

This method has a problem that bubbles easily enter the sample solution when a force bar slip is applied. The mixed bubbles inhibit contact between the probe and the sample and cause false negatives. To avoid this problem, very careful handling of force bar slips is required, and the processing of large samples takes a great deal of time.

Furthermore, the hybridization solution is present in a narrow space covered with a cover slip, and is difficult to disperse, resulting in a bias in the sample concentration, causing false negatives due to insufficient sample volume and false positives due to excessive sample volume. There is a problem. In order for the hybridization solution to be present in the force bar slip region without excess or deficiency, it is necessary to finely adjust the amount of the solution. When two or more probe regions are present on the same carrier and different samples are allowed to react with each probe region on this carrier, if the hybrid solution is excessive, it will be outside the cover slip region, that is, outside the probe region. In this case, the hybridization solution protrudes, causing the contamination of the sample with other probe regions, which causes false positives due to cross-hybridization. In addition, after finishing the hybridization, the force purse slip must be removed for cleaning, but the cover slip is strongly adsorbed on the carrier due to surface tension, and it is difficult to quickly remove it.

As an improved microarray force bar strip, a cover strip that integrates a lid and a frame that is attached to a carrier after immobilizing the probe has been developed (Grace Bio-Labs, Inc, USA). In addition, a method of spotting DNA or antibody in a 96-well microphone opening plate has been developed (Genometrix; Z.ipkin I. BioCentury. 1999, Nov. 15, 7: A9, Mendoza LG et al. Biotechniques. 1999 0ct; 27 (4): 778-80, 782-6, 788). However, operability and reliability over time were not yet satisfactory. Summary of the Invention

The present inventors improve the operability of hybridization and the reliability of analysis data by providing a frame in the detection part of the microarray and holding the sample (test liquid) on the detection part. Was found.

An object of the present invention is to provide a microarray excellent in operability and reliability of analysis data and suitable for processing a large number of samples.

According to the present invention, there is provided a microarray for detecting a plurality of target molecules, a carrier for detecting a target molecule to which a molecule capable of capturing the target molecule (capture molecule) may be immobilized, A microarray having at least one unit comprising a frame for holding a sample on the carrier is provided.

Since the microarray according to the present invention does not have an immobilized lid on the frame, the captured molecule can be immobilized at any time. This makes it easy to discharge and inject the washing liquid. In addition, if the frame is integrated in the microarray according to the present invention in advance, the work of attaching the frame in the detection step is unnecessary, and the work of removing the frame is not required because the frame does not hinder detection of the target molecule. Therefore, the microarray according to the present invention is advantageous in that it has excellent operability.

Since the microarray according to the present invention can stably hold a solution on the detection portion, it is possible to use a probe that can be detected only in a solution such as a molecular beacon probe having excellent reliability and immediacy. Become. Further, in the microarray according to the present invention, even when a plurality of samples are handled on the same carrier, contamination of each sample into another sample can be avoided by clearly dividing a plurality of detection regions by a frame. Therefore, the microarray according to the present invention is advantageous in that a reliable data without erroneous determination can be obtained and a large amount of sample can be processed. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a diagram showing a microarray according to the present invention. Azusa 3 is attached to carrier polystyrene 2. A region surrounded by a frame 3 on the carrier polystyrene 2 is a hybridization region. A is a plan view, B is a sectional view, and C is a perspective view. FIG. 2 is a diagram showing a microarray according to the present invention. A plurality of frames 3 are detachably attached to the carrier polystyrene 2. A region surrounded by a frame 3 on the carrier polystyrene 2 is a hybridization region. A is a plan view, B is a sectional view, and C is a perspective view.

FIG. 3 is a diagram showing a microarray according to the present invention. A plurality of frames 3 integrated by the frame holders 14 are detachably attached to the carrier polystyrene 2 c. A region surrounded by the frame 3 on the carrier polystyrene 2 is a hybridization region.

FIG. 4 shows a microarray set. A is a diagram showing a microarray 5 and a microarray holder 16. B is a diagram showing a microarray set 7 obtained by placing a plurality of microarrays 5 in a box-shaped microarray holder 6. FIG. 5 is a diagram showing an embodiment in which a plurality of microarrays are integrated by fitting concave portions 8 and convex portions 9 provided on the carrier 2 and the frame holder 4 to each other. A microarray set can be obtained by integrating a plurality of microarrays at uneven portions.

FIG. 6 is a view showing an embodiment in which a plurality of microarrays are integrated by a clip-shaped microarray holder 10. A microarray set can be obtained by integrating a plurality of microarrays with the clip 10.

FIG. 7 is a diagram showing a frame according to the present invention. A plurality of frames 3 are integrated by a frame holder 4.

FIG. 8 is a diagram showing the results of HLADRB1 gene tying in the examples. Detailed description of the invention

As used herein, the term “microarray” refers to a carrier in which a group of arranged capture molecules is immobilized on a carrier or a carrier capable of immobilizing the same, and is referred to as a “detection chip” such as a DNA chip or the like. The microarray according to the present invention, which is used to include a carrier for preparation, comprises a carrier for detecting a target molecule and a sample (test liquid) for detecting the presence of the target molecule on a detection portion. And at least one unit consisting of: It preferably has 2 to 10 units, more preferably 2 to 5 units.

In the present invention, “unit” means a detection space separated by a frame. When the microarray has a plurality of units, the carrier constituting each unit can be a common carrier. That is, a plurality of units can be formed by installing a plurality of frames on a common carrier. See, for example, FIGS.

The capture molecule may be immobilized on the carrier in advance.

Target molecules to be detected include antigenic substances (proteins, glycoproteins, amino acids, etc.) or antibodies against them, nucleic acids (DNA, RNA, etc.), peptide nucleic acids, etc., especially during organ or bone marrow transplantation. Of the white blood cell type (HLA) It is considered useful for tying.

The capture molecules for detecting these are antibodies against substances having antigenicity or substances having antigenicity, nucleic acid probes, nucleic acid probes (Hermann T and Patel DJ. Science. 2000 Feb 4; 287 (5454): 820 -5) or a peptide nucleic acid (Beier M and Hoheisel JD.Nucleic Acids Res. 1999 May l; 27 (9): 1970-7), etc., and a molecule such as streptavidin that has the ability to bind to the molecule to be detected. It may be. Preferably, it is a nucleic acid probe. As the nucleic acid probe, a synthetic oligonucleotide, a plasmid / phage, a nucleic acid amplified in a living body such as ssMA, or a nucleic acid fragment amplified in a test tube by PCR or the like can be used.

The capture molecules can be immobilized on the carrier by physical adsorption or chemical bonding. As shown in Schena M et al. Science. 1995 Oct 20; 270 (5235): 467-70, the carrier glass surface was coated with poly-L-Lys and the positive charge of the Lys amino group was observed. A typical example is one utilizing the electrostatic action between the negative charge of the phosphate bond between DNA and DNA. In addition, as is done with the microphone mouth plate (Inouye S and Hondo R. J Clin Microbiol. 1990

Jun; 28 (6): 1469-72), there is also a method of absorbing using a plastic such as polystyrene as a carrier. In chemical bonding, as described in the literature (Za recitation atteo N et al. Anal Biochem. 2000 Apr 10; 280 (1): 143-50), functional groups such as aldehyde groups, carboxyl groups, and amino groups are present on the carrier surface. This is performed by introducing DNA into which a functional group reactive with the functional group is introduced. In addition, the binding ability of biomolecules such as streptavidin and biotin, and antibodies and antigens can also be used.

The capture molecules are preferably arranged on a carrier and immobilized at a high density. The amount of information obtained in a single experiment is dramatically increased due to the high density of the probe, and changes in the state of the living body caused by environmental changes, diseases, administration of drugs, etc. It can be captured as an image. Probe densification is described, for example, in Lipshutz RJ et al. Nat Genet. 1999 Jan; 21 (l Suppl): 20-4 and Schena M et al. Science. 1995 Oct 20; 270 (5235): 467-70. Can be performed according to the method described in It is sufficient that the frame constituting the unit has a height sufficient to hold the test liquid on the detection portion, preferably 3 mm to 15 mm, more preferably 5 mm to 1 O mm It is. The microarray according to the present invention is provided with a frame. If the frame and the carrier are integrated in advance, there is no need to attach the frame before detection. In addition, the detection of the target molecule is not hindered by the frame, so that the removal operation is not required. Therefore, it is advantageous in that the operation procedure can be simplified.

The frame can be removable if required. Specifically, the frame and the carrier can be reversibly bonded with a silicon gasket or the like.

In the microarray according to the present invention, the frame may have one or more projections or depressions, and the carrier may have depressions or projections corresponding to the projections or depressions of the frame. The frame and the carrier can be detachably fixed by fitting the projection or the recess of the frame with the corresponding recess or the projection of the carrier.

In the microarray according to the present invention, a plurality of units may be integrated by a frame holder. Preferably, a plurality of units may be integrated in the opening of the frame. Such an embodiment is as shown in FIG. A plurality of units may be integrated on one carrier so as to be arranged in a line.

The shape of the unit is preferably a rectangular parallelepiped, a cube, or a similar shape suitable for immobilizing the captured molecules efficiently and at high density. The purpose is to prevent the test liquid from scattering and evaporating to the unit as necessary. The bottom surface of a conventional microplate is a circle, and when a plurality of independent regions are arranged in parallel, capture molecules are immobilized between the regions. There were regions where the capture molecules could not be created, and the capture molecules could not be arranged efficiently. In the microarray according to the present invention, the aligned capture molecules can be immobilized on the carrier at high density, and the immobilized portion can be efficiently arranged, so that one microarray can process many samples at a time. This is advantageous in that it can be performed.

In the microarray according to the present invention, a carrier portion where a sample solution is to be present (for example, a probe-immobilized region) is surrounded by a frame, and this frame is moved out of the probe region of the sample solution (for example, a hybridization solution). Block oozing. Therefore, the sample solution can be injected into the carrier portion without worrying that the sample solution spreads to other regions, which is advantageous in that the operation is simplified.

In the space formed by the frame, the sample solution is freely stirred by shaking. As a result, it is possible to prevent the concentration of the target molecule in the solution from being uneven and the bubbles from remaining on the capture molecule. Accordingly, it is advantageous in that more stable hybridization can be performed. Since the frame of the microarray according to the present invention is not covered with the lid on which the frame is fixed, the capture molecules can be immobilized at any time. Therefore, the operation procedure can be made flexible.

In the microarray according to the present invention, since the frame is not covered by the immobilized lid, after reacting the captured molecules with the sample, the detection area is set using an aspire with a nozzle and a dispenser. Can be washed. The aspirator and the dispenser can be, for example, a microplate washer. Therefore, the microarray according to the present invention is advantageous in that the washing can be automated as in the microplate washing. Note that it is also possible to automate the washing process when preparing a microarray.

Further, in the conventional method, the removal of the washing solution after the hybridization was performed by centrifugation or air injection pressure. In this case, the salt of the washing solution after the hybridization often remains. Since the salt remaining on the surface itself fluoresces, it caused non-uniform noise when the scanner sensitivity was increased, hindering the detection of trace samples. In addition, the salt remaining on the probe spot became a large crystal, which sometimes caused a false negative by blocking the excitation light and the fluorescence. The present invention is also advantageous in that detection can be performed while the detection solution is retained, and such a decrease in detection sensitivity is avoided.

In the present invention, a plurality of hybridization regions can be formed by arranging frames as necessary. The fact that a plurality of nodipase regions are physically separated by a frame indicates that highly homologous genes such as the human leukocyte antigen genes HLA-A and HLA-B are detected on the same carrier. However, it is very effective to avoid cross-hybridization caused by contamination of each gene amplification product into the other hybridization region. Detecting multiple genes from the same sample on the same microarray avoids erroneous determination due to a combination error of each gene sequence information that can occur due to a notation error on the container, etc., compared to using multiple containers. This is advantageous in that it can be performed. Even when a plurality of specimens are handled on the same microarray, mixing of each specimen into another specimen can be avoided by clearly dividing the plurality of hybridization areas by a frame. An example of this would be the detection of fungi in hygiene inspections. Although the type of detection target (in this case, bacteria) is not enough to use the entire area of a single slide glass with a microarray carrier, multiple areas with the same probe set on a single microarray are used. By setting, the ability to process many samples is improved.

Furthermore, since the microarray of the present invention can stably hold a solution on a probe, the function can be exhibited only in a solution such as a molecular beacon probe if a device capable of measuring with a frame is used. Inappropriate props can be used. In diagnosis, high reliability and immediacy are required, and it is important to prevent false negatives due to contamination of nucleic acid amplification products with reagents before the gene amplification reaction and to shorten the detection time. Ishi guro T et al. Nucleic Acids Res. 1996 Dec 15; 24 (24): 4992-7, as shown in Orizu E et al. Molecular & Cellular Probes. Probes that emit signals by hybridization (hybrid recognition probes), such as the molecular beacon shown in 1998 Aug; 12 (4): 219-26, do not require sample labeling and do not require further washing. Since detection is possible, the detection system is very simple and the risk of contamination is low. Therefore, the hybrid recognition probe is expected to be applied in the diagnostic field. Application as a microarray probe has been attempted, but molecular beacons can be detected in a dry state as pointed out in Steemers FJ. Et al. Nature Biotechnology. 2000 Jan; 18 (l): 91-4. However, it is difficult to dry with conventional microarrays, and molecular beacons cannot be used as microarray probes because drying is indispensable for scanning. Similarly, it is expected that the conventional method will also be difficult for oligonucleotides linked at once. The microarray container according to the present invention is advantageous in that a hybrid recognition probe excellent in reliability and immediacy can be used since detection can be performed while a liquid is retained on a prop.

In the microarray of the present invention, a removable lid can be used if necessary. The lid prevents evaporation of the hybridization solution in the hybridization. In particular, it is possible to prevent the hybridization solution from scattering outside the container during the transfer of the container. Evaporation of the hybridization solution is a problem because it changes the salt concentration of the hybridization solution, making it impossible to obtain the predetermined hybridization conditions. Also, the scattering of the hybridization solution containing the nucleic acid amplification product leads to serious inspection errors when it contaminates the amplification system. Therefore, the lid can be easily removed while maintaining the required hermeticity for operational reasons during the immobilization of the probe, injection of the sample onto the carrier surface during hybridization, and the washing process after hybridization. desirable. The required tightness is sufficient to avoid evaporation that can cause large fluctuations in the liquid volume during hybridization and does not have to be strong enough to prevent the liquid from leaking out. Therefore, the lid can be a simple seal.

If necessary, a lid or frame can be provided with an inlet or outlet for the hybridization solution to automate a series of reactions.

The carrier portion, frame, and, in some cases, the lid for detecting the target molecule according to the present invention are not particularly limited as long as they have a material having an impermeable surface, but when the detection is performed from the bottom of the microarray, it is preferably transparent. Materials can be used. Examples of transparent materials include glass, plastics such as polystyrene and polypropylene, and processed products obtained by chemical treatment of these materials as required, and processed products coated with polymers such as nitrocellulose and polyacrylamide. . When a fluorescent label is used for detection, the detection sensitivity can be increased by coating with aluminum or coloring the material black. Preferably, ultraviolet rays (Zouali M and Stollar BD. J Immunol Methods. 1986 Jun 10; 90 (1): 105-10) are used to attach plastics such as polystyrene or polypropylene to water-soluble molecules such as proteins and nucleic acids. And a strong acid-strong base (US Pat. No. 5,712,383). The thickness of the microarray is not particularly limited, but is preferably such that it does not affect the light transmittance, and is generally about 0.5 mm to 2 mm.

According to a preferred embodiment of the microarray according to the present invention, a carrier for detecting a target molecule to which a molecule capable of capturing the target molecule (capture molecule) may be immobilized, and a frame for holding the sample on the carrier are provided. Target molecule having multiple units consisting of Provided is a microarray for detecting the number of units, wherein a plurality of units are integrated by a frame holder, and a plurality of units are formed on a common carrier.

According to a more preferred embodiment of the microarray according to the present invention, a carrier for detecting a target molecule to which a molecule capable of capturing the target molecule (capture molecule) may be immobilized, and a frame for holding a sample on the carrier are provided. A microarray for detecting a target molecule, comprising 2 to 10 units consisting of: a unit formed on the same carrier, and a unit formed by a frame holder at an opening of the frame. An integrated microarray is provided.

According to a most preferred embodiment of the microarray according to the present invention, a carrier for detecting a molecule of interest to which a molecule capable of capturing the molecule of interest (capture molecule) may be immobilized, and a sample held on a carrier A microarray for detecting a target molecule, comprising 2 to 10 units comprising a frame, a plurality of units being formed on a common carrier, and a plurality of units being formed at an opening of the frame. Unit is integrated by a frame holder, one or more projections or depressions are provided on the frame, and a depression or projection is provided on the carrier corresponding to the projection or depression of the frame, A microarray is provided in which the frame is detachably fixed to the carrier by fitting the convex or concave portion of the frame with the concave or convex portion of the corresponding carrier.

According to the present invention, there is provided a microarray set in which a plurality of microarrays according to the present invention are integrated. A plurality of microarrays can be integrated by a microarray holder. The microarray holder may be any one that can align and fix a plurality of microarrays, and may be, for example, a box-shaped holder (see Fig. 4) or a clip-shaped holder (see Fig. 6). A plurality of microarrays can also be integrated by fitting one or more protrusions and / or recesses provided on the microarray (see FIG. 5). The uneven portion may be present on both the carrier and the frame holder, or may be present on either of them. The microarray set integrated by the uneven portion may be further supported by a microarray holder (for example, a box-shaped holder). The number of microarrays included in the microarray set is preferably 2 to 12. According to another aspect of the present invention, there is provided a frame for holding a sample on a carrier for detecting a target molecule of a microarray, wherein the frame is used by being immobilized on the carrier. A frame is provided. This frame may be detachable from the carrier part.

This frame may also be provided with one or more convex portions or concave portions corresponding to one or more concave portions or convex portions provided on the carrier. In this case, the frame can be detachably fixed to the carrier by fitting the convex or concave portion of the frame with the corresponding concave or convex portion of the carrier.

Hereinafter, a detection method in the case where the sample is a nucleic acid will be described.

The sample can be a PCR amplified nucleic acid sample. As a nucleic acid label, a fluorescent substance such as Cy3, a radioisotope, or a substance having an affinity for another substance such as hapten-dibiotin such as DNP can be used. These labels can be prepared by using a primer whose terminal is labeled with an arbitrary labeling substance, or by introducing the labeling substance into a monomeric nucleic acid used for amplification in advance. When DNP or biotin is used, it is necessary to add a substance (enzyme-labeled antibody or fluorescently-labeled streptavidin) that enables detection of these chemicals at the time of detection.

When the probe is not immobilized on the microarray according to the present invention, the probe is immobilized on the detection portion.

Next, the sample is poured into the microarray according to the present invention, and the target nucleic acid is hybridized with the probe. Remove and wash the sample if necessary.

After the hybridization, the target nucleic acid is detected. Even when detection is performed by chemiluminescence or color development, a stable result can be obtained because the reaction solution is stably maintained on the probe.

In addition to the above, by examining the amount and activity of a protein translated from mRNA, the state of a cell can be directly known. In this case, different antigen Z antibodies can be spotted on the same carrier, and various proteins can be detected at once by immunological methods such as ELSA (Mendoza LG et al. Biotechniques. 1999 0ct; 27 (4 ): 778-80, 782-6, 788). It can also be used to analyze interactions between proteins (Emili AQ and Cagney G. Nat Biotechnol. 2000 Apr; 18 (4): 393-7).

Example

An embodiment of a biomolecule detection container according to the present invention will be described with reference to FIG.

A microarray container was prepared in which a frame was attached on a flat polystyrene plate so that the hybridization area was 152 mm ² (19 mm × 8 mm). 15 types of human leukocyte antigen gene DRB 1 typing probes (# 1: DRB 2801 J, # 2: DRB1002J, # 3: DRB7004, # 4: DRB 1004, # 5: DRB5703J, # 6DRB3705J, # 7: DRB 10X YJ , # 8: DRB 1006, # 9: DRB70J, # 10: DRB 1007, # 11: DRB 1008J, # 12: DRB 1003NJ, # 13: DRB 1005, # 14: DRB7003, # 15: c ommon It was synthesized according to the method described in Kawai S et al. Eur JI employed unogenet. 1996 Dec; 23 (6): 471-86. This probe was spotted by a frame on the area formed on the polystyrene plate, and solidified by physical adsorption.

The partial sequence of allyl * 1101 of the DRB1 gene was amplified by PCR according to the method described in Kawai S et al. Eur J I uno genet. 1996 Dec; 23 (6): 471-86. However, the 5 'end of the primer was labeled with fluorescein instead of the biotin described in the literature to obtain a fluorescein-labeled amplification product.

The PCR product was purified, diluted with 1 OxSSC to a final concentration of 1 OnM, and used as a solution for hybridization.

Inject 200 / L of hybridization solution into the probe-immobilized area, 45. Hybridization was performed at C for 2 hours. The lid was closed with a seal during the hypride treatment.

The hybridization solution was removed, and the plate was washed three times with 1 × SSC.

After washing, the fluorescence value of each probe spot was measured using a fluorescence microscope. As shown in Table 1, fluorescence was detected with the probes (# 5, # 7, # 12) specific for DRB 1 * 1101 and the common allyl probe (# 15).

As described above, the container according to the present invention enables gene detection and typing. It became clear.

Claims

The scope of the claims

1. A microarray for detecting the target molecule, a carrier for detecting the target molecule on which a molecule capable of capturing the target molecule (capture molecule) may be immobilized, and a sample held on the carrier A microarray having at least one unit comprising a frame for performing the operation.

2. The microarray according to claim 1, wherein the capture molecule is immobilized on a carrier.

3. The microarray c according to claim 1 or 2, wherein the carrier is made of a transparent material.

4. The microarray according to any one of claims 1 to 3, wherein the carrier is glass, plastic, silicon, or a processed product thereof.

5. The microarray according to any one of claims 1 to 4, wherein the unit further has a removable lid.

6. The microarray according to any one of claims 1 to 5, wherein the unit is a cube, a rectangular parallelepiped, or a shape similar thereto.

7. The microarray according to any one of claims 1 to 6, wherein the height of the frame is 3 mm to 15 mm.

8. The microarray according to any one of claims 1 to 7, wherein the frame is detachable from the carrier.

9. The microarray according to any one of claims 1 to 8, comprising a plurality of units.

10. The microarray according to claim 9, having 2 to 10 units.

11. The microarray according to any one of claims 1 to 10, wherein the capture molecule is a nucleic acid, a peptide nucleic acid, an antibody, or an antigen.

12. The microarray according to any one of claims 1 to 11, wherein the target molecule is an HLA gene.

13. A microarray set, wherein the plurality of microarrays according to any one of claims 1 to 12 are integrated.

14. The microarray set according to claim 13, wherein a plurality of microarrays are integrated by a microarray holder.

15. One or more microarrays provided on a microarray The microarray set according to claim 13, wherein the microarray set is integrated by fitting a convex portion and / or a concave portion of the microarray.

16. A frame for holding a sample on a carrier for detecting a target molecule of a microarray, wherein the frame is used by being immobilized on the carrier.

17. The frame according to claim 16, which is detachable from a carrier.