JP4845307B2 - Detection array using a three-dimensional substrate - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
  The present invention is a detection array for detecting a test substance in a biological sample using a specific affinity substance.As a specific affinity substanceProbeThree-dimensional substrateThe present invention relates to a probe array immobilized on a surface.
[0002]
[Prior art]
A detection method utilizing a biological specific binding reaction, for example, a nucleic acid detection method utilizing hybridization is generally used widely. Many apparatuses for performing hybridization in such a method have been developed.
[0003]
For example, Corning provides aminosilane-coated CMT-GAPS as a slide glass dedicated to DNA microarrays. This slide glass is a flat glass slide of a certain standard, that is, about 7.5 cm × about 2.5 cm, and is used as a substrate for a probe array. The user uses various kinds of nucleic acid probes specific to the nucleic acid to be detected on the surface thereof, separately immobilized. The means using such a glass slide has advantages such as being cheaper than other means and easy to immobilize the probe, but uneven reaction in the hybridization reaction is a problem. It has become.
[0004]
On the other hand, a gene chip that arranges probe DNAs on a semiconductor substrate at high density^TMAre provided by Affymetrix (see US Pat. No. 5,143,854, etc.). The gene chip TM is a kind of nucleic acid probe array in which many kinds of probe DNAs are separately solid-phased at high density on one side of a 2 cm × 2 cm flat semiconductor substrate by a photolithography technique. In US Pat. No. 5,843,767, a plurality of through-holes having a diameter of about 5 μm to about 2 mm are formed on a semiconductor substrate having a thickness of about 10 μm to about 500 μm, and probe DNA is attached to each through-hole. A nucleic acid probe array having an increased surface area by being solid-phased on the inner wall is disclosed.
[0005]
As described above, the conventional nucleic acid probe arrays each use a flat plate-like substrate. However, in such a conventional flat substrate, in a nucleic acid probe array preparation process and a hybridization reaction process, a large amount of reaction solution, for example, so that the amount of liquid does not evaporate at least during the required reaction time, for example, Since a reagent such as a specimen, a washing solution, and a labeled target DNA is required, it is not suitable for processing a large amount of substrates.
[0006]
Further, the flat substrate has a small portion of the surface of the substrate, and at least one substrate surface is not used for the solid phase. In addition, the flat substrate requires an extra amount to bring the entire necessary reaction part into contact with the specimen, and the dispensing accuracy in each reaction part is likely to fluctuate, so reaction results with low reproducibility can be obtained. There is sex.
[0007]
On the other hand, as an example of increasing the surface area, a bead with a diameter of several μm is mixed in a solution containing a specific type of specific affinity binding reagent to solidify the reagent and react with the sample in a reaction vessel. Also known is a bead-shaped substrate that measures the total light output, such as fluorescence, for beads that are floating or settled on the bottom of the container, but position information on the substrate surface cannot be obtained. Mistakes in the results of inspection items can occur.
[0008]
In addition, there is no provision of an appropriate method for differentiating a plurality of substrates to be reacted and / or measured corresponding to each of a plurality of different analytes, which can lead to sample mis-mixing. Also, when performing a plurality of different inspections simultaneously or successively, it is difficult to distinguish and grasp each inspection situation.
[0009]
[Problems to be solved by the invention]
In view of such circumstances, the present invention can reduce the amount of reagents required in each work step, and can carry out processing without causing reaction unevenness at any position of the detection array. An object of the present invention is to provide a detection array that can be easily and stably processed at a time without unevenness of reaction.
[0010]
[Means for Solving the Problems]
  As a result of diligent research, the inventors have conventionally solidified a specific affinity substance on a flat plate.FlatChip array3D solid substrateBased on the bold idea of changing to a spherical shape, we have developed a means to solve the above problems and achieve the purpose. That is,The present inventionAt least one three-dimensional substrate and a plurality of types of specific affinity substances that specifically bind to a plurality of types of test substances are provided on the three-dimensional substrate.Specific locationOn the surfacePartiallySolid phase specific affinity probe andA position detection marker for discriminating the arrangement of the specific affinity probe on the substrate surfaceAnd an array for detection.
[0011]
DETAILED DESCRIPTION OF THE INVENTION
[Specific affinity substance]
Specific affinity substances of the present invention include, for example, substances containing nucleic acids such as genes, antigens, antibodies, allergens, hormones and enzymes.
[0012]
[Nucleic acid probe array]
The nucleic acid probe array of the present invention comprises a three-dimensional substrate and a nucleic acid probe 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, a curved body is particularly preferable, and a true sphere is most preferable.
[0013]
[Three-dimensional substrate]
The three-dimensional substrate constituting the probe array of the present invention means one or a plurality of substrates constituting a three-dimensional surface. The three-dimensional surface has a three-dimensional region having a level that can be detected by a known detection technique, and preferably, at least a part thereof has a curved surface.
[0014]
[Granular substrate]
The granular substrate constituting the nucleic acid probe array is about 4.2 × 10^-3mm³To about 6.5 × 10⁴cm³A granular substrate having a volume of The form of the granular substrate used in the present invention is not particularly limited as long as it is granular, and specifically, a cube, a rectangular parallelepiped, a polyhedron, etc., and a shape having a curved surface in a part thereof, such as a sphere, a spheroid, and others Including a rotating body. Moreover, those forms may be hollow, and further, a form including a curved surface in at least a part obtained by dividing the curved body into two parts, three parts, or four parts. A preferred feature is a true sphere. However, the present invention is not limited to these. Here, the term “curved surface” used in the above-mentioned cubic base and thus granular base is a true sphere (FIG. 1), an elliptical rotator (FIG. 2), a hemisphere (FIG. 3), and a bowl (FIG. 4). , Egg-shaped, bowl-shaped, cigar-shaped, rice-shaped, rocket-shaped, conical, cylindrical (including elongated fiber bodies), and any shape surrounded by curved surfaces such as streamlines, and one derived from these curved surfaces Including, but not limited to, a shape having a partial curved surface. However, a rotating body such as a true sphere or an elliptical rotating body is preferable because of the ease of the manufacturing process.
[0015]
As used herein, the term “granular” is a word that indicates the general form of a relatively small substrate, the specific shape and size are as shown in this specification, and a conventional substrate having a plate shape. Is a term that means that a nucleic acid probe is partially solid-phased and has a curved surface in at least one part.
[0016]
The volume of the substrate is about 4.2 × 10^-3mm³To about 6.5 × 10⁴cm³Preferably about 0.065 mm³To about 4.19 cm³More preferably about 0.52 mm.³To about 0.52 cm³It is. For example, in the case of a true sphere, the diameter may be a maximum of 50 cm depending on the number of types of specific affinity substances to be immobilized, but in practice, it may be 200 μm to 5 cm, preferably 500 μm to 2 cm, preferably 1 mm To 1 cm is most preferred. It is also possible to simultaneously use carriers of different sizes in the same reaction system. 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, when the substrate has a cross-sectional diameter of 2 mm or less, particularly a minute portion such as less than 1 mm, A long and narrow fiber body or rod-like body that has a long enough length (for example, 1 to 50 cm) is reduced to a spherical shape, a conical shape, a cylindrical shape, or the like by appropriately bending it into an irregular shape or a spiral shape 3 It may be a cube with a dimensional structure (for example an average diameter of 200 μm to 1 cm and / or a longitudinal part of 1 mm to 5 cm).
[0017]
The granular substrate used in the present invention can have various required properties (rigidity, airtightness) by mixing glass, plastic material, magnetic material, gelatin, rubber, protein, silicon, etc. alone, or by appropriately mixing, encapsulating, and laminating. Properties, surface roughness, specific gravity, light permeability, solubility, charge, water absorption, hue, brightness), but is not limited thereto.
[0018]
The granular substrate can be formed by means known per se according to the material. For example, when glass is used as a 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 that are granulated 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-like or cylindrical shape can also be used. . In order to maintain the cubicity of the substrate, the manufacturing cost can be reduced by coating, winding, planting, etc., a material suitable for solid-phase nucleic acid to various rigid members as a skeleton. Good.
[0019]
Further, the surface of the granular substrate is preferably smooth to an appropriate level for immobilizing the nucleic acid probe. In addition, the surface of the substrate can be subjected to a surface treatment suitable for solidifying the nucleic acid probe, for example, a surface treatment such as poly-L-lysine treatment, aminosilane treatment, and oxide film treatment.
[0020]
A major feature of the nucleic acid probe array of the present invention is that a granular substrate having a curved surface in at least one part is used as a substrate for solid-phase nucleic acid probes. The curved surface has a remarkably large surface area compared to a flat substrate of the same volume that has been conventionally used. That is, the granular substrate used in the present invention has a plate-like substrate with the same diameter of 4r.²The surface area of the sphere is 4πr compared to²Therefore, a surface area about three times as large can be obtained. Accordingly, it becomes possible to arrange a larger number of probe DNAs on a small substrate, and this can improve the packaging density, thereby achieving resource saving. In addition, as will be described later, the amount of sample necessary for performing a hybridization reaction can be reduced, and at the same time, an equally stable reaction can be performed in all the nucleic acid probe arrays used.
[0021]
In addition, when the nucleic acid probe is immobilized on the substrate, it is preferable to provide some positioning marker in order to determine the arrangement of the nucleic acid probe on the substrate surface. The method of applying such a marker includes providing a plurality of protrusions, recesses or grooves, labeling the nucleic acid probe itself, and providing a label that is not accompanied by irregularities on the substrate surface and can be optically detected. It is possible to use means such as performing. It is also possible to attach a plurality of markers to one substrate. In particular, it is preferable to attach three markers because each immobilization position for any of the same or different nucleic acid probes can be easily recognized.
[0022]
[Nucleic acid probe]
The nucleic acid probe to be solid-phased in 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. In addition, oligonucleotides, PCR products, and the like are used as the probe DNA, but are not limited thereto. The length of the base chain may be appropriately determined according to the nucleic acid to be detected. When a nucleic acid probe is synthesized, it can be synthesized in advance prior to the solid phase on the substrate, or can be synthesized on the substrate.
[0023]
[Solidification method]
The probe supply means for immobilizing a desired nucleic acid probe on the substrate is modified so that any means known per se can be applied to the curved surface of a three-dimensional substrate or granular substrate, particularly a spherical surface, depending on the material of the substrate. Can be done. For example, in order to place a nucleic acid probe on a substrate with a curved surface processed, a combination of two methods, a photosolid phase method and a spotting method, can be used quantitatively with respect to a curved partial region. The nucleic acid probe can be solid-phased. Furthermore, in the present invention, it is important to combine a probe supply means and a means for relatively moving various substrates (preferably a granular substrate, particularly a spherical substrate) along a three-dimensional surface (preferably a curved surface, particularly a spherical surface). .
[0024]
The photo-solid phase method is an application of photolithographic technology. The substrate is masked in advance, the reaction site is irradiated with light to perform deprotection, and the nucleic acid probe is specifically immobilized only on the deprotected portion. It is a method to do. When a plurality of kinds of nucleic acid probes are immobilized on an actual solid phase, the above operation is repeated. The granular substrate suitable for the optical solid phase is particularly preferably one using silicon. For example, when a spherical silicon substrate is used, after the polycrystalline silicon is heat-treated, it is naturally dropped inside a glass tube or the like. Obtained by cooling and single crystallization (see US Pat. No. 5,955,776).
[0025]
For light irradiation, a continuous exposure apparatus for a granular substrate can be used (refer to JP-A-11-121368). This continuous exposure apparatus exposes a plurality of granular substrates that are continuously conveyed without stopping the movement of the substrates. Here, in order to realize exposure along the curved surface of the substrate, it is preferable to fix the position of the substrate so that the substrate can be rotationally controlled. Therefore, for example, the fixing of the substrate to the granular substrate holder is performed by a reduced pressure or a centrifugal force generated by the rotation of the annular member of the apparatus. It is possible to adjust the position of the granular substrate by individually operating at least three ultrasonic actuators and rotating the held spherical substrate. By further incorporating a part for performing the step of reacting and binding the nucleic acid probe to the substrate after exposure, and then a part for washing and drying the granular substrate, this apparatus can be used as an optical solid phase immobilization apparatus for the nucleic acid probe. It is possible to use. Note that it is possible to rotate or shift not only the spherical substrate but also the light source or the light irradiation site.
[0026]
On the other hand, the spotting method refers to a method in which a probe is solid-phased on the substrate by an inkjet method in which probe DNA is jetted or a pin method in which probe DNA is attached by contact with a pin. The spotting apparatus used to manufacture the array of the present invention is to rotate or shift the inkjet head and pins on the probe DNA discharge side, or rotate or change the spherical substrate side with respect to the substrate having a spherical surface. It is desirable to be able to perform these and combinations thereof. In addition, such an apparatus could continuously process not only one substrate but also a plurality of substrates.
[0027]
Further, when the nucleic acid probe is immobilized on the substrate, it is desirable to apply some positioning marker to the granular substrate in order to accurately arrange the nucleic acid probe on the surface of the substrate and to advantageously perform detection. Such markers are provided with a plurality of protrusions, recesses or grooves, labeled on the nucleic acid probe itself, and labeled with optical detection without accompanying irregularities on the substrate surface, etc. This is possible by using the following means. FIGS. 5 and 6 show an example in which the protrusion 1 is asymmetrically or applied to both poles, FIG. 7 shows an example in which the recess 2 is applied symmetrically and to both poles, and FIG. 8 shows an example in which the groove 3 is applied asymmetrically. However, the present invention is not limited to this. The number of the protrusions is not limited to one or two as shown in the figure. For example, a sufficient number of protrusions may be provided so that contact between the surface of the substrate and the inner wall of the container is prevented. In this case, it may be easy to grasp the arrangement of each nucleic acid probe by making the size or shape of only a specific protrusion different from other protrusions. In addition, as a method for grasping the arrangement of the nucleic acid probe on the substrate, in addition to providing projections or grooves, any signal generating means capable of emitting a signal different from the detection label may be used. In addition, when a plurality of types of nucleic acid probes are immobilized on different positions on the same substrate or on each surface portion of different substrates, the signal generating means is different for each type of nucleic acid probe. It is preferable to set various hues, pigment amounts, magnetic amounts and the like so as to obtain a signal amount or a signal pattern. By using such a signal generating means, it is not necessary to provide the above-mentioned position detection marker on the substrate, and it is not necessary to control the position of the solid phase on the substrate, so that the production of the detection array of the present invention is simple and low cost. There is also an advantage of making it.
[0028]
Further, when performing the solid phase of the nucleic acid probe, it is desirable to fix and hold each substrate and to precisely adjust the relative position with respect to the apparatus. Therefore, it is necessary to perform precise positioning by rotating or shifting the substrate, and to securely hold the granular substrate at the adjusted position. For this reason, the granular substrate of the present invention can be provided with one or a plurality of protrusions or recesses as shown in FIGS. By arranging these, positioning becomes easy, and it is possible to easily rotate or horizontally move the base body from that portion. Further, the size and shape of the plurality of protrusions and recesses described here can be changed.
[0029]
According to the present invention, when performing a plurality of types of specific affinity binding reactions, the positional relationship between the substrate and the specific affinity binding reagent can be grasped in the solid phase step and / or the measurement step. When a plurality of different inspections are continuously performed, it is easy to distinguish and grasp each inspection state. Furthermore, in order to distinguish a plurality of different specimens, a plurality of different test substrates (one or more of each) can be transmitted, reflected, refracted, or polarized by a desired combination of different colors, densities, and brightness. Since multiple types of substrates with different optical properties such as can be provided for each sample, by executing reaction, measurement, result output, etc. by associating the individual samples with the optical properties of the substrate Also, it has a revolutionary advantage of high throughput, ease of use, reliability and the like that hardly cause a difference even if a plurality of different specimens are processed simultaneously or continuously.
[0030]
[how to use]
Examples of use of the nucleic acid probe array of the present invention are shown below. That is, an example of a method for detecting DNA using the nucleic acid probe array of the present invention is shown.
[0031]
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 the DNA probe provided in the nucleic acid probe array and the target DNA.
[0032]
Here, as the target DNA, it is conceivable to use, as a sample, a traceable label molecule, for example, a label labeled with a fluorescent dye, a luminescent dye, or the like, or a substance that can be amplified with a primer labeled with the target molecule. . Labels that can be used include, but are not limited to, Cy3, FITC and biotin. In addition, a method of detecting a change in polarization after hybridization using an unlabeled target DNA can also be used.
[0033]
One preferred use example will be described with reference to FIG. A hybridization reaction is performed by adding the nucleic acid probe array of the present invention and a reagent to one well of a 96-well microtiter plate (FIG. 9). At this time, the nucleic acid probe array includes a true spherical substrate having a diameter of 6 mm and a DNA probe as a nucleic acid probe. The reagent to be added is about 160 μL. After hybridization under appropriate conditions, a washing operation is performed using a plate washer.
[0034]
Next, another usage example will be described with reference to FIG. The nucleic acid probe array of the present invention and a reagent are added to one well of a 96-well microtiter plate, 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 a DNA probe as a nucleic acid probe, and 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. In this way, if a plurality of types of DNA probes corresponding to target DNAs to be detected on a plurality of substrates are individually or combined and solid-phased separately, the production cost per substrate is increased. There is also an advantage that 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 a plurality of substrates be provided with optically or physically labelable labels. The target DNA can be obtained by mixing not only one type of label but also two or more types of target DNA. Further, by providing magnetism to the substrate, it is possible to accumulate a large number of substrates in a part of the well in the cleaning operation, thereby preventing the granular substrate from being attracted. The presence / absence or amount of labeling substance remaining on the array after such a washing operation can be determined by appropriate measuring means and data processing means while classifying the labeling substance on the substrate or by the type of labeling substance. it can.
[0035]
Suitable reaction vessels for hybridization reactions and post-reaction washing operations are versatile, such as microplates with the desired number of multi-holes, such as 96-well microtiter plates, or microtubes from 0.2 μL to 1.5 μL. However, the present invention is not limited to this. The reaction vessel may be tubular like a flow cell or a dispensing nozzle, and has one or more openings having a capillary force that can be surpassed by appropriate pressure from a syringe or the like. You may have a place.
[0036]
The hybridization reaction is generally performed at a constant temperature of 45 ° C. to 65 ° C. for 1 hour to overnight, depending on conditions such as nucleic acid to be detected. It is possible to change the reaction conditions.
[0037]
  Conventionally, when a hybridization reaction is attempted, it is performed in a stationary state using a cover glass for preventing drying. However, when the nucleic acid probe array of the present invention is used, the reaction can be carried out with shaking or stirring in the presence of the present array and sample in a tube or well.That is, since it is possible to perform a hybridization reaction between the substrate and the specimen while stirring in a container,It is possible to avoid the reaction unevenness that is a problem in the prior art.Stable reaction can be carried out. In addition, since the contact frequency between the probe DNA and the target DNA increases, an excellent effect of increasing the hybridization reaction efficiency can be obtained.
[0038]
In addition, if the hybridization reaction and the washing operation after the reaction are performed with a standardized microtiter plate, the washing operation can be performed using a washer for the microtiter plate, and the washing operation from the hybridization reaction. Can be automated or semi-automated. Therefore, by using the nucleic acid probe array of the present invention together with a versatile container, it is possible to further improve the operability and shorten the cleaning time in the cleaning operation.
[0039]
  Furthermore,Since the present invention is smaller than a conventional nucleic acid probe array having the same area, the amount of reagents and test samples can be reduced. That is,By selecting an appropriate reaction vessel based on the diameter of the granular substrate, it is possible to perform the reaction and washing with a small amount of liquid sufficient to cover the granular substrate, and to save resources such as reagents. .It is also possible to subject a large number of nucleic acid probe arrays to the reaction in one container at a time.It is also possible to change the diameter of the granular substrate in accordance with the reaction vessel. For example, when one nucleic acid probe array is used per well, about 6 to 7 mm for 96 wells and about 3 mm for 384 wells can be preferably used. In FIG. 10, the diameter and the number of granular substrates are set to be laminated in two or more layers in the well. However, if they are set to be one layer, the arrangement of the substrates is two-dimensionally arranged. This is more preferable.Moreover, by using a combination of nucleic acid probe arrays of different sizes, it is possible to efficiently use the space in the container, thereby realizing resource saving and reagent saving. .
[0040]
By carrying out the hybridization reaction using a plurality of substrates in the same container as described above, the surface area of the substrate per well can be increased, and therefore resource saving can be achieved. It is also possible to use a combination of substrates having different sizes. By using in this way, it is possible to reduce the required amount of reagent.
[0041]
It is also preferable to use the nucleic acid probe array of the present invention together with the container shown in FIGS. As shown in FIG. 11, the reaction solution can be reduced in volume by making the bottom of the well or the test tube U-shaped. Further, as shown in FIGS. 12 and 13, by arranging a recess or projection corresponding to the recess or projection (or also referred to as a projection) provided on the substrate provided in the nucleic acid probe array of the present invention in the container. It is possible to stably support a spherical substrate in a container, so that the arrangement address of each nucleic acid probe included in the substrate can be obtained even for a substrate having a shape that is easy to roll, such as a spherical shape or a cylindrical shape. Can be easily grasped, and sufficient reaction and washing can be performed without the DNA probe on the substrate contacting the bottom surface of the test tube and hiding. Furthermore, as shown in FIG. 14, the same thing can be obtained by providing magnetism 14 at a specific portion of the base body so as to give partial magnetism partially or unevenly, and disposing a magnet 13 outside the container to be used. In addition, the substrate can be stably fixed in a fixed direction. Examples of a method for arranging a magnetic substance at a specific portion of the substrate include a spotting method and an encapsulation method. The positioning of the substrate shown in FIGS. 12, 13 and 14 can be effectively used for solid-phase treatment and measurement of nucleic acid probes. In particular, according to the positioning method shown in FIG. 14, for example, solid phase treatment or measurement can be performed simultaneously or continuously in a state where one or a plurality of substrates are fixed in a fixed direction on a shallow container. Furthermore, the substrate can be oriented in all directions by intermittently rotating the magnet below the container, moving the magnet to the side of the U-shaped bottom, or rotating the magnet on the side. Therefore, it becomes easy to solidify and measure with a sufficient surface area.
[0042]
【The invention's effect】
  BaseTraditionalFrom flat plateThree-dimensional surface with a three-dimensional surfaceBy immobilizing a plurality of types of specific affinity substances as shapes, it became possible to increase the surface area without requiring a large amount of processing liquid such as sample, reagent, and washing liquid. As a result, the mounting density can be improved, and more probes can be fixed. Therefore, resource saving is possible.
[0043]
  Furthermore, since the positional relationship with the specific affinity binding reagent can be grasped, when a plurality of different examinations are executed simultaneously or successively, it is easy to distinguish and grasp each examination situation.
[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 cross-sectional view showing a bowl-shaped spherical substrate.
FIG. 5 is a side view showing a true spherical spherical base body having a protrusion.
FIG. 6 is a side view showing a true spherical spherical substrate having a protrusion.
FIG. 7 is a cross-sectional view showing a true spherical spherical substrate having a recess.
FIG. 8 is a side view showing a true spherical spherical substrate having grooves.
FIG. 9 is a view showing a preferred use example of the present nucleic acid probe array.
FIG. 10 is a view showing a preferred use example of the present nucleic acid probe array.
FIG. 11 is a diagram showing an example of a preferable container shape for using the nucleic acid probe array.
FIG. 12 is a view showing an example of a preferable container shape for using the nucleic acid probe array.
FIG. 13 is a diagram showing an example of a preferable container shape for using the nucleic acid probe array.
FIG. 14 is a view showing an example of a preferable container shape for using the nucleic acid probe array.
[Explanation of symbols]
1. Projection part Recess 3. Groove 4. 4. Reaction vessel Granular substrate
6). DNA probe 7. Reaction solution 8. Labeled target DNA
9. Washing nozzle 10. Magnetic granular substrate
11. First labeled target DNA 12. Second labeled target DNA
13. Magnet 14. Magnetic part

Claims (5)

At least one granular substrate having a curved surface in a part thereof and a plurality of types of specific affinity substances that specifically bind to a plurality of types of test substances, respectively , over the three-dimensional region of the granular substrate. A detection array comprising a specific affinity probe immobilized on at least a part of the curved surface , and at least three position detection markers for discriminating the arrangement of the specific affinity probe on the surface of the substrate.   The detection array according to claim 1, wherein the position detection marker is a protrusion or a recess.   The detection array according to claim 1 or 2, wherein the position detection marker is a signal generating means capable of generating a signal amount or a signal pattern that is different for each type of the probe. The array for detection according to any one of claims 1 to 3 , wherein magnetism is provided ubiquitously with respect to a specific portion of the substrate. The detection array according to any one of claims 1 to 4 , wherein the specific affinity substance is selected from the group consisting of a substance containing a nucleic acid such as a gene, an antigen, an antibody, an allergen, a hormone, and an enzyme. Array.

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