JP2002181819A - Detection array using solid base body - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a detection array capable of reducing an amount of reagents required in respective working processes, performing treatment without causing any uneven reaction in any position of the detection array, and stably treating a great number of tested substances easily at once without any reaction variation. SOLUTION: This detection array is provided with at least one solid base body and a specific affinity probe formed by immobilizing a plurality of kinds of specific affinity substances specifically binding with a plurality of kinds of tested substances onto the surface of the solid base body.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a detection array for detecting a test substance in a biological sample using a specific affinity substance, and is used, for example, for detecting a nucleic acid. The present invention relates to a nucleic acid probe array. More specifically, the present invention relates to a nucleic acid probe array in which a nucleic acid probe specific to a nucleic acid to be detected is immobilized on the surface of a granular substrate.

[0002]

2. Description of the Related Art A detection method utilizing a biological specific binding reaction, for example, a nucleic acid detection method utilizing hybridization is generally widely used. Many devices for performing hybridization in such a method have also been developed.

For example, Corning provides aminosilane-coated CMT-GAPS as a slide glass dedicated to DNA microarrays. The slide glass is a flat slide glass of a certain standard, that is, about 7.5 cm × about 2.5 cm, and is used as a substrate of the probe array. The user uses various types of nucleic acid probes specific to the nucleic acid to be detected on the surface thereof, separately immobilizing them. Means using such a slide glass are inexpensive compared to other means, and have advantages such as easy immobilization of a probe,
The problem is uneven reaction in the hybridization reaction.

On the other hand, a probe DN is provided on a semiconductor substrate.
Gene chip to arrange A at high density ^TMIs Affyme
(Affymetrix) (US Patent No.
No. 5,143,854). GeneChipTM is
Photolithography on one side of a 2 cm x 2 cm planar semiconductor substrate
Various types of probes D at high density by lithography technology
A kind of nucleic acid probe array in which NAs are separately immobilized
You. Also, U.S. Pat.
Is about 10 μm to about 500 μm on a semiconductor substrate,
A large number of through holes with a diameter of about 5 μm to about 2 mm are formed,
By immobilizing the probe DNA on the inner wall of each through-hole,
Thus, a nucleic acid probe array having an increased surface area is disclosed.
ing.

[0005] As described above, each of the conventionally used nucleic acid probe arrays uses a flat plate-shaped substrate. However, in such a conventional plate-like substrate, in the nucleic acid probe array preparation process and the hybridization reaction process, a large amount of reaction solution, In addition, it requires reagents such as a specimen, a washing solution, and a labeled target DNA, so that it is not suitable to process a large amount of substrates.

[0006] Further, the flat substrate has little use of the surface of the substrate, and at least one substrate surface is not used for the solid phase. In addition, the plate-like substrate requires an extra amount to bring the entire required reaction portion into contact with the sample, and the dispensing accuracy in each reaction portion is likely to fluctuate.
Possibility of obtaining reaction results with low reproducibility.

On the other hand, as an example of increasing the surface area, beads having a diameter of several μm were mixed with a solution containing a specific affinity binding reagent of a specific type to immobilize the reagent and reacted with a sample in a reaction vessel. Later, a bead-shaped substrate that measures the total light output such as fluorescence for beads floating or settling on the bottom of the container in a free direction is also known, but it is not possible to obtain positional information on the substrate surface. Since it is not possible, the result of the inspection item may be confused.

[0008] In addition, there is no suitable method for distinguishing a plurality of substrates to be reacted and / or measured from each other corresponding to each of a plurality of different analytes. In addition, when performing a plurality of different inspections simultaneously or consecutively, it is difficult to distinguish and grasp each inspection status.

[0009]

In view of such circumstances, the present invention reduces the amount of reagent required in each operation step and treats the reagent without any uneven reaction at any position of the detection array. It is another object of the present invention to provide a detection array which can carry out a large number of samples at a time and can stably process the reaction without unevenness.

[0010]

Means for Solving the Problems As a result of diligent research, the present inventors have based on a bold idea of changing a chip-shaped array, on which a specific affinity substance is conventionally immobilized on a flat plate, into a spherical shape, based on a bold idea. We have developed means to solve the above problems and achieve the purpose. That is, a specific affinity probe in which at least one three-dimensional substrate and a plurality of types of specific affinity substances each of which specifically binds to a plurality of test substances are immobilized on the surface of the three-dimensional substrate. And a detection array comprising:

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION [Specific Affinity Substance] The specific affinity substance of the present invention includes, for example, substances containing nucleic acids such as genes, antigens, antibodies, allergens, hormones and enzymes.

[Nucleic acid probe array] The nucleic acid probe array of the present invention comprises a three-dimensional substrate and nucleic acid probes immobilized on the surface of the three-dimensional substrate. Preferably, the three-dimensional substrate is granular, more preferably has a curved surface in at least one portion, particularly preferably a curved surface, and most preferably a true sphere.

[Three-dimensional substrate] The three-dimensional substrate constituting the probe array of the present invention means one or a plurality of three-dimensional substrates constituting a three-dimensional surface. The three-dimensional surface is three-dimensionally provided with a region at a level detectable by a known detection technique, and preferably has at least a part of a curved body.

[Granular Substrate] The granular substrate constituting the nucleic acid probe array has a particle size of about 4.2 × 10 ⁻³ mm ³ to about 6.
It is a granular substrate having a volume of 5 × 10 ⁴ cm ³ . The form of the granular substrate used in the present invention may be granular, and specifically, cubes, cuboids, polyhedrons, and the like, and a part having a curved surface, such as a sphere, a spheroid, and others Including rotating body. In addition, their form may be hollow, and may be a form in which at least a part obtained by dividing the curved body into two, three, or four also includes a curved surface. The preferred form is a true sphere. However, it is not limited to these. here,
The term “curved body” used in the above-mentioned cubic substrate, and hence the granular substrate, is a true sphere (FIG. 1), an ellipsoidal rotator (FIG. 2), a hemisphere (FIG. 3), a bowl shape (FIG. 4), an oval shape , Bales, cigars, rice, rockets, cones, cylinders (including slender fibrous bodies), and any other shapes that are surrounded by curved surfaces such as streamlines, and partial curved surfaces derived from those curved surfaces Including, but not limited to. However, a rotator such as a true sphere or an elliptical rotator is preferable from the viewpoint of ease of the manufacturing process.

As used herein, the term "granular" refers to the general shape of a relatively small substrate, and the specific shape and size are as shown in the present specification, and are plate-like. This term is used in comparison with a conventional substrate, and means a term having a curved surface in at least one part where a nucleic acid probe can be partially immobilized.

The volume of the substrate is about 4.2 × 10 ⁻³ mm
³ may be about 6.5 × ¹⁰ 4 cm ^3, preferably from about 0.065 mm ³ about 4.19 cm ^3, more preferably from about 0.52 mm ³ to about 0.52cm ^3. For example, in the case of a true sphere, its diameter can be up to 50 cm depending on the number of specific affinity substances to be immobilized, but in practice it may be from 200 μm to 5 cm,
Preferably from 500 μm to 2 cm, most preferably from 1 mm to 1 cm. It is also possible to use carriers of different sizes simultaneously in the same reaction system. Further, in the case of a substrate having a curved surface portion and a non-curved surface portion such as a cone or a cylinder, for example, in the case of a substrate having a cross-sectional diameter of 2 mm or less, particularly a minute portion such as less than 1 mm, A combination of elongated fibrous bodies or rods whose length is made sufficiently long (for example, 1 cm to 50 cm) is reduced to a spherical, conical, or cylindrical shape by appropriately bending an irregular or helical shape. It may be a cube with a three-dimensional structure (for example, an average diameter of 200 μm to 1 cm and / or a longitudinal part of 1 mm to 5 cm).

The granular substrate used in the present invention is glass,
Various necessary properties (rigidity, airtightness, etc.) can be obtained by mixing, enclosing and laminating plastic materials, magnetic materials, gelatin, rubber, proteins, silicon, etc., alone or appropriately.
It can be produced by giving surface roughness, specific gravity, light transmittance, solubility, charge, water absorption, hue, and brightness), but is not limited thereto.

The granular substrate can be formed by means known per se in accordance with the material. For example,
When glass is used as the material, a desired shape can be obtained by molding or polishing. A plastic material is processed into a spherical shape by injection molding or molding, but is not limited to these methods.
In addition, spheres, polygonal particles, and irregularly shaped particles formed by a method of forming a solid phase from a liquid phase, such as coacervation, or a method of winding and solidifying a fibrous material in a pill shape or a cylindrical shape and the like can also be used. . In order to maintain the cubicity of the substrate,
The production cost may be reduced by coating, winding, planting, or the like a material suitable for immobilizing nucleic acids on various rigid members serving as a skeleton.

It is preferable that the surface of the granular substrate is as smooth as appropriate for immobilizing the nucleic acid probe. The surface of the substrate can be subjected to a suitable surface treatment for immobilizing the nucleic acid probe, for example, a poly-L-lysine treatment, an aminosilane treatment, and an oxide film treatment.

A significant feature of the nucleic acid probe array of the present invention is that a granular substrate having a curved surface in at least one portion is used as a substrate for immobilizing a nucleic acid probe on a solid phase. The curved surface has a remarkably large surface area as compared with a conventionally used flat substrate having the same volume. That is, in the granular substrate used in the present invention, the surface area of the sphere is 4π in comparison with the area of the plate-like substrate having the same diameter of 4r ^2.
because it is r ^2, the surface area of about 3 times can be obtained. Therefore,
It is possible to arrange a larger number of probe DNAs on a small substrate, thereby improving the packaging density, thereby achieving resource saving. In addition, as will be described later, the amount of sample required for performing the hybridization reaction can be reduced, and at the same time, a stable reaction can be performed equally in all the nucleic acid probe arrays used.

When the nucleic acid probe is immobilized on the substrate, it is preferable to provide some kind of positioning marker in order to determine the arrangement of the nucleic acid probe on the surface of the substrate. Methods for applying such a marker include providing a plurality of protrusions, recesses, or grooves, labeling the nucleic acid probe itself, and labeling without optically detectable irregularities on the substrate surface. It is possible to use means such as performing. It is also possible to attach a plurality of markers to one base. In particular, it is preferable to attach three markers because each immobilized position of a plurality of the same or different nucleic acid probes can be easily recognized.

[Nucleic acid probe] The nucleic acid probe immobilized on the nucleic acid probe array of the present invention is a nucleic acid having a sequence complementary to the nucleic acid to be detected, and includes DNA and RNA. Probe D
Oligonucleotides, PCR products, and the like are used as NA, but are not limited thereto. The length of the base chain may be appropriately determined according to the nucleic acid to be detected. When synthesizing a nucleic acid probe, it can be synthesized in advance prior to the solid phase on the substrate, or can be synthesized on the substrate.

[Method for Immobilizing a Solid Phase] A probe supplying means for immobilizing a desired nucleic acid probe on the substrate may be any known means depending on the material of the substrate, and may be any means known per se, for example, a curved surface of a three-dimensional substrate or a granular substrate, particularly This can be done by making modifications to be applicable to spherical surfaces. For example, to arrange a nucleic acid probe on a substrate having a curved surface processed,
The nucleic acid probe can be quantitatively immobilized on a curved partial region by appropriately combining the two methods of the optical solid phase method and the spotting method. Further, in the present invention, the probe supply means and the various substrates (preferably granular substrates, particularly spherical substrates) are formed in a three-dimensional surface (preferably a curved surface,
In particular, it is important to combine means for relatively moving along a spherical surface).

The photo-solid phase method is an application of the photolithographic technique, in which a substrate is masked in advance, and only the reaction site is irradiated with light to perform deprotection. In a solid phase. When immobilizing multiple types of nucleic acid probes on the actual solid phase,
Repeat the above operation. Granular substrates suitable for the optical solid phase are particularly preferably those using silicon.For example, when a spherical silicon substrate is used, after polycrystalline silicon is heat-treated, it falls naturally inside a glass tube or the like. It is obtained by cooling and single crystallization (U.S. Pat.
955,776).

For the light irradiation, a continuous exposure apparatus or the like for a granular substrate can be used (see Japanese Patent Application Laid-Open No. 11-121368). This continuous exposure apparatus exposes a plurality of continuously transported granular substrates without stopping the movement of the substrates. Here, in order to realize exposure along the curved surface of the substrate, it is preferable that the position of the substrate is fixed by a structure capable of controlling rotation. Thus, for example, the fixing of the substrate to the granular substrate holder is performed by decompression or by centrifugal force generated by rotation of the annular member of the device. By individually operating at least three ultrasonic actuators and rotating the held spherical substrate, it is possible to adjust the position of the granular substrate. In this device,
Furthermore, by further incorporating a site for performing a step of reacting and binding a nucleic acid probe to a substrate after exposure, and then performing a site for washing and drying the granular substrate, it is possible to use the device as a nucleic acid probe photo-solidification device. It is possible. In addition,
It is possible to rotate or shift not only the spherical substrate but also the light source or the light irradiation site.

On the other hand, the spotting method refers to a method in which a probe is solid-phased on the substrate by an ink jet method of spraying probe DNA or a pin method of attaching probe DNA by contact with a pin. The spotting apparatus used to manufacture the array of the present invention rotates or shifts an ink jet head and a pin on the probe DNA discharge side with respect to a substrate having a spherical surface, or rotates or shifts the spherical substrate side. It is desirable to be able to perform these combinations and combinations thereof.
Further, such an apparatus would be capable of continuously processing not only one substrate but also a plurality of substrates.

When the nucleic acid probe is immobilized on the substrate, some positioning marker is applied to the granular substrate to accurately arrange the nucleic acid probe on the surface of the substrate and to perform the detection advantageously. Is desirable. Such a marker may be provided with a plurality of projections, depressions or grooves, labeled on the nucleic acid probe itself, and may be labeled without any irregularities on the substrate surface and capable of optical detection. It is possible by using the means of (1). 5 and 6 show an example in which the projections 1 are provided asymmetrically or at both poles, FIG. 7 shows an example in which the recesses 2 are provided symmetrically and at both poles, and FIG. 8 shows an example in which the grooves 3 are provided asymmetrically. However, the present invention is not limited to this. The number of the projections is not limited to one or two as shown in the figure. For example, a sufficient number of projections may be provided so as to prevent contact between the surface of the base and the inner wall of the container. In this case, the size or shape of only the specific protrusion may be different from that of the other protrusions to facilitate understanding of the arrangement of each nucleic acid probe. In addition, as a method of grasping the arrangement of the nucleic acid probe on the substrate, other than providing a projection or a groove, any signal generating means that can emit a signal different from the detection label may be used, such as fluorescence, dye, magnetism, and the like.
Any one may be used, and further, when a plurality of types of nucleic acid probes are immobilized on different positions of the same substrate or on respective surface portions of different substrates, the signal generating means is provided for each type of nucleic acid probe. It is preferable to set variously the hue, the dye amount, the magnetic amount, and the like so as to obtain different signal amounts or signal patterns. By using such a signal generating means, it is not necessary to provide the above-mentioned marker for position detection on the substrate, and it is not necessary to control the position of the solid phase on the substrate. There is also the advantage of doing.

When the solid phase of the nucleic acid probe is performed,
It is desirable to fix and hold each substrate and precisely adjust the relative position to the device. Therefore, it is necessary to perform precise positioning by rotating or displacing the substrate, and to reliably hold the granular substrate at the adjusted position. For this reason, the granular substrate of the present invention can be provided with one or more projections or depressions as shown in FIGS. By arranging them, positioning becomes easy, and rotation or horizontal movement of the base starting from that portion can be easily performed. In addition, the plurality of protrusions and recesses described here are:
It is possible to change the size and shape.

According to the present invention, when performing a plurality of types of specific affinity binding reactions, it is possible to grasp the positional relationship between the substrate and the specific affinity binding reagent in the solid phase step and / or the measurement step. In the case where a plurality of different inspections are performed simultaneously or continuously, it is easy to distinguish each inspection status. Further, in order to distinguish between a plurality of different analytes, a plurality of different test substrates (one or more of each) are subjected to light transmittance, reflectance, refraction, and polarization by a desired combination of different colors, densities, and brightness. Since a plurality of types of substrates having different optical properties such as the above can be provided for each sample, the reaction, measurement, result output, etc. are performed by associating the individual sample with the optical properties of the substrate. ,
There is also an epoch-making advantage of high throughput, ease of use, high reliability, and the like, in which even if a plurality of different specimens are processed simultaneously or consecutively, it is unlikely that mixing will occur.

[Method of Use] An example of the use of the nucleic acid probe array of the present invention will be described below. That is, an example of a method for detecting DNA using the nucleic acid probe array of the present invention will be described.

First, according to the method described above, a desired DNA probe is immobilized on the granular substrate to obtain a nucleic acid probe array. A hybridization reaction is performed between a DNA probe provided in the nucleic acid probe array and a target DNA.

Here, as the target DNA, a traceable labeled molecule, for example, one labeled with a fluorescent dye, a luminescent dye, or the like, or one that can be amplified with a primer labeled with the target molecule, is used as a sample. Can be considered. Labels that can be used include, but are not limited to, Cy3, FITC, and biotin. Also,
A method of detecting a change in polarization after hybridization using an unlabeled target DNA can also be used.

One preferred use example will be described with reference to FIG. A nucleic acid probe array of the present invention and a reagent are added to one well of a 96-well microtiter plate to perform a hybridization reaction (FIG. 9). At this time, the nucleic acid probe array includes a true spherical substrate having a diameter of 6 mm and DNA probes as nucleic acid probes. The reagent to be added is about 160 μL. After hybridization under appropriate conditions, a washing operation is performed using a plate washer.

Next, another example of use will be described with reference to FIG. 96-well microtiter plate 1
The nucleic acid probe array and the reagent of the present invention are added to the wells, and a hybridization reaction is performed (FIG. 10). At this time, the nucleic acid probe array includes a true spherical substrate having a diameter of 2 mm and DNA probes as nucleic acid probes.
About 8 nucleic acid probe arrays are added per well.
Here, it is assumed that one or two or more different nucleic acid probes are immobilized on each of the eight nucleic acid probe arrays. Thus, a plurality of types of D corresponding to a target DNA to be detected with respect to a plurality of substrates are obtained.
If the NA probes are individually or individually combined and immobilized separately, there is also an advantage that the manufacturing cost per substrate can be reduced. The reagent to be added is about 100 μL. After hybridization under appropriate conditions, a washing operation is performed using a plate washer washing nozzle. In this case, it is desirable that the plurality of substrates be provided with a labelable optical or physical label. The target DNA may be a mixture of not only one type of label but also two or more types of target DNA. In addition, by providing the substrate with magnetism, a large number of substrates can be accumulated in a part of the well in the cleaning operation, thereby preventing the granular substrate from being sucked. The presence or absence or amount of the labeling substance remaining on the array after such a washing operation can be determined by a suitable measuring means and data processing means while classifying the labeling substance on the position on the substrate or the type of the labeling substance. it can.

A reaction vessel suitable for the hybridization reaction and the washing operation after the reaction is a microplate having a desired number of multiwells such as a 96-well microtiter plate, or a microtube having a size of 0.2 μL to 1.5 μL. , But is not limited thereto. Further, the reaction vessel may be a tubular one such as a flow cell or a dispensing nozzle, and one or more openings having a capillary force such that the inside of the vessel can be overcome by a suitable pressure from a syringe or the like. It may have some places.

In general, the hybridization reaction is carried out at a constant temperature of about 45 ° C. to about 65 ° C. for 1 hour to overnight with a target DNA.
It is possible to change the reaction conditions according to the conditions such as the nucleic acid to be detected.

Conventionally, when a hybridization reaction is attempted, it is performed in a stationary state using a cover glass for preventing drying. However, if the nucleic acid probe array of the present invention is used, the present array and a sample can coexist in a tube or well, and the reaction can be performed under shaking or stirring. As a result, it is possible to avoid uneven reaction which is a problem in the related art. Probe DN
Since the frequency of contact between A and the target DNA increases, an excellent effect of increasing the efficiency of the hybridization reaction can be obtained.

If the hybridization reaction and the washing operation after the reaction are performed on a standardized microtiter plate, the washing operation and the like can be performed using a washer for a microtiter plate. Automatic or semi-automatic cleaning is possible. Therefore, by using the nucleic acid probe array of the present invention together with a versatile container, the operability in the washing operation and the washing time can be further shortened.

Further, by selecting an appropriate reaction vessel based on the diameter of the granular substrate, it is possible to carry out the reaction and washing with a small amount of liquid enough to cover the granular substrate, Resources can be saved. It is also possible to change the diameter of the granular substrate according to the reaction vessel. For example, if one present nucleic acid probe array is used per well, about 6 to 7 mm for 96 wells, 3
About 3 mm can be preferably used for 84 wells. Also,
In FIG. 10, the diameter and the number of the granular substrates are set for laminating two or more layers in the well, but if they are set to be one layer, the arrangement of each substrate is grasped two-dimensionally. It is more preferable because it is possible.

As described above, by performing a hybridization reaction using a plurality of substrates in the same container,
The surface area of the substrate per well can be increased, and thus resource saving can be achieved. It is also possible to use substrates having different sizes in combination. By using in this way, it is possible to reduce the required amount of the reagent.

FIG. 1 shows the nucleic acid probe array of the present invention.
It is also preferable to use with the containers shown in 1 to 14.
By making the bottom of a well or a test tube U-shaped as shown in FIG. 11, the reaction solution can be reduced in volume. In addition, as shown in FIGS. 12 and 13, a concave portion or a convex portion corresponding to a concave portion or a convex portion (also referred to as a projecting portion) provided on a substrate provided in the nucleic acid probe array of the present invention is provided in a container. In addition, it is possible to stably support a spherical substrate in a container, and thus, even for a substrate having a shape that easily rolls, such as a spherical or cylindrical shape, the arrangement address of each nucleic acid probe provided on the substrate. As a result, sufficient reaction and washing can be performed without hiding the DNA probe on the substrate in contact with the bottom surface of the test tube. Furthermore, as shown in FIG. 14, by providing magnetism 14 at a specific portion of the base to partially or unevenly provide partial magnetism, and by arranging magnet 13 outside the container to be used, the same applies. Furthermore, it is possible to stably fix the base in a certain direction. Examples of a method for disposing a magnetic material at a specific portion of the base include a spotting method and a sealing method. The positioning of the substrate shown in FIGS. 12, 13 and 14 can be effectively used also in the case of solid-phase treatment and measurement of a nucleic acid probe. In particular, according to the positioning method shown in FIG. 14, for example, solid-phase processing and measurement can be performed simultaneously or continuously with one or a plurality of substrates fixed on a shallow vessel in a fixed direction. Further, the substrate can be oriented in any direction by intermittently rotating the magnet below the container, or moving the magnet to the side of the U-shaped bottom surface, or rotating the magnet on the side. Therefore, solid-phase formation and measurement with a sufficient surface area are facilitated.

[0042]

According to the present invention, since a plurality of types of specific affinity substances are immobilized on a substrate from a flat plate to a cubic shape having a curved surface, the surface area can be reduced without requiring a large amount of a sample, a reagent, and a processing solution such as a washing solution. It became possible to increase. As a result, the mounting density can be improved, and more probes can be fixed. Therefore, resource saving can be achieved.

Since the hybridization reaction between the substrate and the sample can be carried out while stirring in the vessel, it is possible to carry out a stable reaction while avoiding uneven reaction. Further, since the present invention is smaller than a conventional nucleic acid probe array having the same area, it is possible to reduce the amount of reagents and test samples. Further, a large number of nucleic acid probe arrays can be subjected to the reaction at one time in one container. Also, by using a combination of nucleic acid probe arrays of different sizes,
The space in the container can be used efficiently, thereby realizing resource saving and reagent saving.

[Brief description of the drawings]

FIG. 1 is a side view showing a true spherical base.

FIG. 2 is a side view showing a spheroidal spherical substrate.

FIG. 3 is a side view showing a hemispherical spherical substrate.

FIG. 4 is a sectional view showing a bowl-shaped spherical substrate.

FIG. 5 is a side view showing a true spherical spherical base having a projection.

FIG. 6 is a side view showing a true spherical base body having a projection.

FIG. 7 is a cross-sectional view showing a true spherical spherical base having a concave portion.

FIG. 8 is a side view showing a true spherical spherical substrate having a groove.

FIG. 9 is a view showing a preferred use example of the present nucleic acid probe array.

FIG. 10 is a diagram showing a preferred use example of the present nucleic acid probe array.

FIG. 11 is a view showing an example of a preferred container shape for using the present nucleic acid probe array.

FIG. 12 is a view showing an example of a preferred container shape for using the present nucleic acid probe array.

FIG. 13 is a view showing an example of a preferred container shape for using the present nucleic acid probe array.

FIG. 14 is a view showing an example of a preferable container shape for using the present nucleic acid probe array.

[Explanation of symbols]

1. Protrusion 2. Recess 3. Groove 4. Reaction vessel
5. 5. Granular substrate DNA probe 7. Reaction solution 8. 8. Labeled target DNA Cleaning nozzle 10. Magnetic granular substrate 11. First labeled target DNA 12. 12. second labeled target DNA Magnet 14. Magnetic part

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) G01N 33/566 G01N 1/28 J 37/00 102 C12N 15/00 FF term (reference) 2G052 AA28 AB20 AD46 DA01 DA02 DA08 DA22 EB11 EC00 FC03 FC12 FD06 FD11 FD20 GA11 GA25 GA30 HA02 HB02 HC40 JA01 JA04 JA06 JA07 JA09 JA13 JA16 4B024 AA11 BA80 CA01 CA09 CA11 HA12 4B029 AA07 BB20 CC03 FA12 4B063 QA01 QQ42 QQ52 QR32 QR35 QR55 QR32 QR35 QS36

Claims (8)

[Claims] An at least one three-dimensional substrate and a plurality of types of specific affinity substances, each of which specifically binds to a plurality of types of test substances, are immobilized on the surface of the three-dimensional substrate. An array for detection comprising an affinity probe. 2. A detection array comprising: a granular substrate having a curved surface in at least one portion; and a nucleic acid probe partially and quantitatively immobilized on the surface of the granular substrate. 3. The detection array according to claim 1, wherein the specific affinity probe has a plurality of types of specific affinity substances immobilized on the same substrate, and An array for detection arranged for each type at a specific position on the surface. 4. The nucleic acid probe array according to claim 1, wherein the specific affinity substance is immobilized by a photo-immobilization means. 5. The nucleic acid probe array according to claim 1, wherein the specific affinity substance is immobilized on a solid phase by spotting means. 6. The detection array according to claim 1, wherein the volume of the substrate is about 0.52 mm ³ to about 0.5.
An array for detection that is 2 cm ³ . 7. The detection array according to claim 3, wherein different kinds of specific affinity substances are solid-phased on the surface of the substrate over the rotation trajectory. 8. The detection array according to claim 1, wherein the shape of the substrate is a true sphere, a cone, or a cylinder.