JP2007121185A - Plate for discriminating living body sample and its manufacturing method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for manufacturing a plate for discriminating living body sample precisely and simply in a case that the fine flow channel or through-hole of the plate is made fine in width or depth (width, 100 μm or below) or the occupying ratio of the through-hole to the total area of the plate becomes high (5% or above), and to provide the plate for discriminating living body sample manufactured by this manufacturing method. <P>SOLUTION: The plate for discriminating a living body sample having a groove and a non-piercing hole or the through-hole formed thereto and having a thickness of 1 mm or above is formed by bonding a plate 1a, to which the groove and the through hole are simultaneously formed by hot emboss processing using a mold, and a plate 1b to which the non-piercing hole or the through-hole is formed by either one of injection molding or transfer molding and machining, by an adhesive or hot-press bonding. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a biological sample discriminating plate for analyzing a biological sample by moving a DNA, protein or other biological sample in a buffer and detecting its transport reaction.

  When a general biological sample is considered, DNA and protein exist largely. In recent years, with the rapid development of molecular biology, the involvement of genes in various diseases has been understood fairly accurately, and attention has been focused on medical treatments targeting genes.

  With regard to DNA, SNPs (abbreviation of single nucleotide polymorphism, which is generally translated as “single nucleotide polymorphism” and is a general term for the difference of one code (one base) in a gene) are currently attracting attention. The reason for this is that the classification of SNPs can predict the prevalence of many diseases and the effects and susceptibility of each individual to drugs, and there are absolutely no humans on the planet who have the same SNPs, even if they are parents and siblings. This is because it is considered that the individual can be completely identified from not doing so.

  At present, as a method for examining SNPs, sequencing (determination of a base sequence) in which a base sequence of DNA is directly read from the end is most commonly used. There are several reports on the sequencing method, but the most common method is dideoxy sequencing (Sanger method). Sequencing is based on a technique that separates and distinguishes the difference in length of one base by denaturing polyacrylamide gel electrophoresis with high resolution or capillary electrophoresis in any method including the Sanger method. It is established.

  Another method is affinity ligand capillary electrophoresis. Affinity ligand capillary electrophoresis uses intermolecular affinity, particularly specific affinity in the ecosystem (enzyme and substrate, antigen and antibody affinity, etc.) to give specificity to separation. Specifically, When an affinity ligand that specifically recognizes the base sequence is added to the electrophoresis solution in the capillary tube and the sample is electrophoresed, only the molecular species that interact in the sample mixture will change in the moving speed. The analysis is performed with attention (for example, see Patent Document 1).

  On the other hand, proteins exist in cells, tissues, and biological fluids, regulate biological activities, supply energy to cells, synthesize important substances, maintain biological structures, and communicate between cells and intracellular information. Involved in transmission. Currently, it has become clear that proteins have multiple functions depending on the various environments, the presence of other interacting proteins, and the degree and type of modification that the protein has undergone.

  Proteins are made by connecting 20 types of amino acids in order according to gene instructions (sequence information), and the types are said to be tens of millions. If you know the sequence of the gene, what amino acid is what? You can get information on whether they are connected in order. A complete set of proteins made from the genes (genomes) of organisms is called a proteome, and now the analysis of the proteome has been actively promoted after the nucleotide sequence of the human genome has been deciphered.

  Such functional analysis of proteins requires not only identification and characterization, but also biochemical assays, protein-protein interaction studies, protein networks, and intracellular and extracellular signaling elucidation. Research on this protein function uses a variety of techniques, including enzyme assays, yeast two-hybrid assays, chromatographic purification, information tools and databases, etc. Protein discrimination by electrophoresis is particularly important. It is a technique.

  Then, as in electrophoresis, the transport reaction obtained when moving liquids such as samples, analytes, buffers, and reagents in capillary tubes is detected, and analysis, discrimination, determination, etc. of the samples are performed. There are various reports regarding the transport and orientation of the liquid when performed (for example, Patent Document 2 and Patent Document 3). In the analysis of these biological samples, a method using an electrophoresis apparatus is widely used, and as a consumable item, a capillary tube or a plastic plate in which a fine channel is formed is used.

As a manufacturing method of the plastic plate in which the fine channel is formed, for example, there is a method of forming by injection molding using a mold or machining, but a mold that can be obtained with high reproducibility in both dimensions and shape. Is preferable (for example, Patent Document 4). In addition, the mold used for this molding sometimes has a groove formed by machining to form a fine flow path. However, the finer the groove width becomes, for example, 100 μm or less, the more accurately the mold is processed. Since it becomes difficult, at present, a method of forming a mold by electroforming plating using LIGA technology is used. In addition, the through-hole formation of the mold can be formed by providing a pin.
JP 7-311198 A Special Table 2000-513813 JP-T-2001-523341 JP 2000-310613 A

  However, the fine flow passages and through holes of the plastic plate described above are further refined in the width and depth of the flow passage (width of 100 μm or less), and the proportion of the through holes in the total plate area has increased (5%). In this case, the electroforming mold (the material is nickel) has a higher coefficient of thermal expansion than HPM38 (SUS type), which is a constituent material of the mold, so that it enters the cavity in addition to the high mold temperature during molding. Due to the cyclic action of the inflowing resin, sliding occurs between the electroforming mold and the mold constituent material for each molding cycle, and as a result, variation in shape accuracy (channel width, channel depth, through hole) ) Becomes larger.

  In addition, since there are a plurality of pins for forming the through holes, a problem of insufficient filling of the flow path portion by the pins occurs. Therefore, in the injection molding, it was impossible to mold the fine flow path and the through hole at the same time.

  In the present invention, the fine flow path and the through hole of the plastic plate are further refined in the width and depth of the flow path (width of 100 μm or less), and the ratio of the through hole to the total plate area is increased (5%). In this case, an object is to provide a method for producing a plastic plate accurately and easily.

  A plastic plate in which a groove and a non-through hole or a through hole are formed, and the plate thickness is 1 mm or more, and a plate 1 in which the groove and the through hole are simultaneously formed by hot embossing using a mold. The non-through hole or the plate 2 in which the through hole is formed is joined by an adhesive or thermocompression bonding by any of injection molding, injection molding, and machining.

  Furthermore, the plastic plate serves as a biological sample discriminating plate for discriminating biological samples.

  Further, of the surfaces constituting the plastic plate, on the first surface on which the groove and the non-through hole or the through hole are formed, the groove formed on the first surface is on the first surface. The surface area occupied by the non-through holes formed in the first surface on the first surface, the surface area occupied by the through holes formed in the first surface on the first surface, Is 5% or more of the total surface area of the first surface.

  Further, the plate 1 has a thickness of 0.3 mm to 1 mm.

  Further, in the plastic plate, a convex portion that is continuous with the wall surface that forms the non-through hole or the through hole is formed on the surface of the plate 2 to be joined to the plate 1.

  Further, the convex portion has a cylindrical shape having an inner diameter equal to the inner diameter of the through hole of the plate 2.

  Further, the convex portion is located at an equal distance from the central axis of the through hole of the plate 1 and is formed so as to be accommodated in the through hole of the plate 1.

  Further, the plate 1 is formed by forming a groove only on one surface, and bonding the surface of the plate 1 on which the groove is not formed and the plate 2.

  Further, in a plastic plate having a groove and a non-through hole or a through hole and having a plate thickness of 1 mm or more, a plate 1 in which the groove and the through hole are simultaneously formed by hot embossing using a mold. And a plate 2 in which a non-through hole or a through hole is formed by one of injection molding, injection molding, and machining, and a convex portion is formed on one surface is joined by an adhesive. In the plastic plate manufacturing method of manufacturing the plastic plate, the adhesive is applied to the surface of the plate 1 opposite to the surface where the groove and the through hole are formed, and the plate 1 and the plate 2 are bonded to each other. It has the process of joining by.

  According to the present invention, even if there are a plurality of fine shapes and through holes in the plate surface, it can be accurately produced. Moreover, it is useful as what can respond | correspond to the thick thing whose plate | board thickness is 1 mm or more. As described above, a biological sample analysis plate capable of accurately producing a fine shape and a through hole can be provided.

(Embodiment 1)
First, the configuration of the biological sample analysis plate according to the first embodiment will be described with reference to FIGS. FIG. 1 is a diagram seen from the flow path forming surface of the biological sample analysis plate according to the first embodiment.

  Eight patterns 2 shown in FIG. 2 are formed in a radial pattern on the plate 1 in the first embodiment, and DNA discrimination of eight specimens can be performed simultaneously.

  As shown in FIG. 1, the outer shape of the plate 1 according to the first embodiment is a square of 8 centimeters and four corners are chamfered, and one corner thereof is further chamfered.

  Further, a hole 4 is provided, which is designed so that the pattern location can be specified by making the outer shape asymmetric. The reason why the plate 1 has such an outer shape is that the plate is attached to an optical reader (not shown) to facilitate positioning when detecting the above-described fluorescence and absorbance. The plate 1 is composed of a plate 1a having a plate thickness of 0.5 mm and a plate 1b having a plate thickness of 1.5 mm, and the plate 1a and the plate 1b are bonded using an adhesive. The material of the plate 1 is an acrylic resin and has a thickness of 2 mm. Grooves are formed on the flow path forming surface, and a sealed flow path is formed by adhering an acrylic film 21 having a thickness of 50 μm.

  Reference numeral 5 denotes an axis of the main plate, and a hole 3 is provided around the axis 5 for fixing the main plate to the rotating portion of the optical reading device.

  2 is a buffer injection port that also serves as an air hole for injecting a DNA conjugate as a buffer, 7 is a buffer injection part for temporarily holding the injected buffer, and 8 is a DNA sample. A sample injection port 9 serving also as an air hole for injection is a sample injection part for temporarily holding the injected DNA sample. The buffer injection part 7 and the sample injection part 9 have similar shapes, and a peripheral cross section is shown in FIG.

  FIG. 3 is a view showing a cross section in the vicinity of the buffer injection portion and the sample injection portion, and the hatched portion is the plate 1. 19 is the buffer injection port 6 and the sample injection port 8, and 20 is the buffer injection unit 7 and the sample injection unit 9. 21 is the above-described film, and a sealed flow path is formed by pasting the groove so as to cover it. Incidentally, the flow path forming surface is the lower part.

  Returning again to FIG. 2, the sample injection unit 9 is connected to the sample quantitative fractionation unit 10, and the buffer injection unit 7 is also connected to the quantitative fractionation unit 10 via the flow path 11. Further, the quantitative sorting unit 10 is scanned with an air hole 13 for venting air, an overflow chamber 14 for quantifying the injected DNA sample by centrifugal force, and the above-described optical reader during electrophoresis. A flow path 12 is connected, and an air hole 15 is connected to an end of the flow path 12. Further, 16 is used for an excess buffer agent that cannot be held in the buffer storage portion to flow in. An overflow chamber is installed. In the present embodiment, the width of the flow path 11 is 300 μm, the depth is 50 μm, the width of the flow path 12 is 200 μm, and the depth is 50 μm.

  Next, a method for manufacturing the plate 1 will be described.

  The plate 1 is formed by bonding a plate 1a having a plate thickness of 0.5 mm and a plate 1b having a plate thickness of 1.5 mm, and the plate 1a is formed by fine flow paths and through holes. Molding was performed, and the plate 1b was configured only by through holes, so that molding was performed by injection molding.

  First, the plate 1 a is a photomask having a shape pattern corresponding to the buffer injection part 7, the sample injection part 9, the sample fixed quantity collection part 10, the flow path 11, the flow path 12, the air holes 13 and 15, and the overflow chambers 14 and 16. Is used to pattern the ultra-thick film resist. Next, Ni electroforming is performed using this as a mother mold to form a molding die. Next, acrylic molding is performed by hot embossing using this mold. Since this mold is formed with a plurality of buffer injection parts 7 and sample injection parts 9, the resin biting of the mold tends to occur when releasing after hot embossing.

  Here, the height of the buffer injection part 7 and the sample injection part 9 of the mold and the plate thickness of the plate 1a were changed to confirm the state of resin biting during hot embossing. The result is shown in FIG. The horizontal axis represents the plate thickness of the plate 1a, and the vertical axis represents the amount of warpage of the plate 1a after release. When measuring with the optical reading device, when the warp of the plate 1a was set to 50 μm or less and the reference value was set, the thickness of the plate 1a was 0.3 mm to 1.0 mm and the warp was 50 μm or less.

  When the plate thickness of the plate 1a is 0.2 mm or less, since the plate itself is thin, warpage becomes large at the time of mold release. In addition, when the plate thickness of the plate 1a is 1.2 mm or more, the buffer injection part 7 and the sample injection part 9 of the mold easily bite into the resin, and the warpage is increased. Therefore, the plate thickness of the plate 1a is preferably 0.3 mm to 1.0 mm. In the present embodiment, the thickness of the plate 1a is 0.5 mm with the least warpage.

  Next, it was tried whether the plate 1a could be produced using an injection molding machine. First, the ultra-thick film resist is patterned by using a photomask having a shape pattern corresponding to the sample fixed quantity collection unit 10, the flow channel 11, the flow channel 12, the air holes 13 and 15, and the overflow chambers 14 and 16. Next, using this as a mother mold, Ni electroforming was performed to produce a molding die. The buffer injection part 7 and the sample injection part 9 were formed by forming a through hole using a pin. And as a result of shaping | molding, it turned out that the accuracy dispersion | variation in a flow-path shape is large.

  This is because the electroformed mold (nickel is made of nickel) has a higher coefficient of thermal expansion than HPM38 (SUS type), which is a component of the mold, so that the high mold temperature during molding is added to the cavity. Due to the cyclic action of the inflowing resin, sliding occurs between the electroforming mold and the mold components for each molding cycle, resulting in variations in shape accuracy (channel width, channel depth, through hole). Becomes larger. Furthermore, it was found that in addition to variations in accuracy, the flow path portion was insufficiently filled.

  Since there are a plurality of pins, it seems that the pins were hindered when filling the resin, resulting in insufficient filling. In chip fabrication in Patent Document 4, it is considered that fabrication by injection molding was successful because the area ratio in which flow paths and through holes are formed is as small as about 4%. The area ratio in which the flow path and the through hole are formed in this embodiment is about 10%, which is larger than the chip of Patent Document 4. Therefore, when the area ratio in which the flow path and the through hole are formed is 5% or more, it is necessary to form by using the hot embossing described above.

  Then, the manufacturing method of the plate 1b is demonstrated.

  As shown in FIG. 4, the plate 1b includes a buffer injection port 6, a buffer injection unit 7, a sample injection port 8, a sample injection unit 9, and a joining rib 17. Since there is no fine structure, the plate 1b is manufactured by injection molding. went. The mold was manufactured by machining, and a pin was used to form the through hole. The material used was the same acrylic as the plate 1a. The joining rib 17 has a cylindrical shape, and the inner diameter size thereof is equal to the diameter of the buffer injection part 7 and the sample injection part 9. Further, the joining rib 17 is located at an equal distance from the central axis of the buffer injection part 7 and the sample injection part 9 of the plate 1a, and is accommodated in the through hole of the buffer injection part 7 and the sample injection part 9 of the plate 1a. Therefore, the outer diameter size is equal to or smaller than the diameters of the buffer injection part 7 and the sample injection part 9.

  Subsequently, the joining of the plate 1a and the plate 1b will be described with reference to FIG.

  First, a methylene dichloride-based adhesive 18 is applied to the surface of the plate 1a opposite to the surface where the grooves and through holes are formed, and the bonding rib 17 of the plate 1b, the buffer injection portion 7 of the plate 1a, and the sample injection The part 9 is joined. At this time, since the joining ribs 17 are well accommodated in the through holes, there is no need for alignment. In the present embodiment, an adhesive is used. However, it may be bonded by heat fusion or a hot melt sheet.

  Now, an example of specific operations and operations up to the determination of the presence or absence of SNPs (single nucleotide polymorphisms) in the DNA sample described above will be described with reference to FIG.

  First, a DNA sample as a specimen is prepared. Originally, DNA has a double-stranded helical structure, but in the first embodiment, a single-stranded DNA having a length of about 60 bases including a SNPs site to be discriminated is prepared. Since the extraction method and single strand formation are not directly related to the present invention, detailed description thereof is omitted.

Next, a DNA conjugate is prepared as a buffer. A DNA conjugate is obtained by covalently bonding a high-molecular linear polymer to the 5 ′ end of a 6- to 12-base long single-stranded DNA. Furthermore, DNA is a sequence that is complementary to the normal type but not complementary to the mutant type, has a strong binding force to the normal type DNA, and a weak binding force to the mutant type DNA. There are characteristics. In addition, when electrophoresed, the linear polymer bonded to the 5 ′ end is weighted and the migration speed is considerably slow. The “DNA conjugate” described hereinafter has physical properties including a pH buffer that also serves as an electrolyte and a DNA binding force control agent such as MgCl ₂ .

  When sample preparation is complete, the DNA conjugate and DNA sample are injected into the plate. First, the DNA conjugate is dispensed by a pipetter or the like so as to fill the buffer injection portion 7 from the buffer injection port 6. Although the dispensing amount varies depending on the scale of the pattern, in the first embodiment, in order to fill the buffer injection part 7, the flow path 11, the quantitative sorting part 10, and the flow path 12, the DNA conjugate is 3 micron. Liters are needed. The volume of the buffer injection part 7 is formed in advance so as to be slightly larger than 3 microliters.

  Next, the plate 1 is fixed to the rotating part of the optical reading device, and the shaft center 5 is rotated about the shaft. The DNA conjugate dispensed at this time moves toward the outer circumference by centrifugal force. The DNA conjugate in the buffer injection part 7 passes through the flow path 11 and moves to the quantitative fractionation part 10, and further moves to the flow path 12. FIG. 6 shows a state in which the movement of the DNA conjugate is stopped after 2 minutes from the start of rotation at a rotational speed of 4000 rpm. Further, FIG. 7 is an enlarged view of the periphery of the quantitative sorting unit 10.

  After the movement of the DNA conjugate is completed, the liquid level height of the DNA conjugate existing in the buffer injection part 7 and the flow path 12 and the liquid level height of the quantitative sorting part are the same with the axis 5 as the center. It will be on the circumference. However, when the amount of the DNA conjugate injected into the buffer injection unit 7 is less than 3 microliters, the DNA conjugate is not filled into the quantitative fractionating unit 10 or the flow path 12 as shown in FIG. 8a. If it is not filled, electrophoresis cannot be performed.

  On the other hand, in the case of more than 3 microliters, an overflow chamber 16 is provided for the flow of surplus buffer that cannot be held in the buffer reservoir. For this reason, the liquid level height of the DNA conjugate and the liquid level height of the quantitative fractionation part are always on the same circumference with the axis 5 as the center. As shown in FIG. 8 b, the overflow chamber 16 has a hatched portion excluding the overflow chamber 16, that is, the hatched volume of the buffer injection part 7, the flow path 11, the quantitative fractionation part 10, and the flow path 12 is 3 μm. It is arranged in the flow path 12 on the same circumference centering on the center of gravity 5 when it becomes a liter.

  Next, after the DNA conjugate is transported into the channel, the DNA sample is injected into the plate. The DNA sample is dispensed from the sample injection port 8 to the sample injection unit 9 using a pipetter or the like. In the first embodiment, since the DNA sample requires 1 microliter, the volume of the sample injection portion 9 is formed slightly larger than that.

  Next, the plate 1 is fixed to the rotating part, and the shaft center 5 is rotated about the shaft. The DNA sample dispensed at this time moves toward the outer periphery by centrifugal force.

  The DNA sample in the sample injection unit 9 moves to the quantitative sorting unit 10 without generating bubbles by the air holes 13. At this time, the DNA sample moved to the quantitative sorting unit 10 can be quantified by the overflow chamber 14, and the unnecessary DNA sample moves into the overflow chamber 14. FIG. 9 shows a state in which the movement of the DNA sample is stopped after 2 minutes from the start of rotation at 4000 rpm. Further, FIG. 10 is an enlarged view of the periphery of the quantitative sorting unit 10.

  The DNA sample remaining in the quantitative sorting unit 10 by the above operation becomes the final sample for determining the SNPs.

  Next, electrophoresis is performed. A film 21 is affixed to the front surface of the plate on which the flow path is formed, and a voltage can be applied by inserting a needle-like electrode there and inserting it into the interior. In the case of a positive electrode, the electrode is pierced at the end of the flow path 12, and in the case of a negative electrode, the electrode is pierced by the buffer injection part 7. In electrophoresis, when a voltage of several hundred volts is applied between the electrodes, an electric field is generated in the flow path 12 and further in the quantitative fractionation unit 10. To the positive electrode side (A direction in FIG. 9).

  The channel 12 is filled with a DNA conjugate, and the DNA sample is electrophoresed while repeating the binding with the DNA conjugate. At this time, as described above, the normal DNA in the DNA sample has a strong binding force with the DNA conjugate, so the migration speed is slow, and the mutant DNA has a weak binding power, so the migration speed is faster than the normal type. That is, when both the normal type and the mutant type are present in the DNA sample, the normal type DNA and the mutant type DNA are separated, and SNPs can be discriminated.

  As described above, according to the present embodiment, even if there are a plurality of fine shapes and through holes in the plate surface, it can be accurately produced. Moreover, it is useful as what can respond | correspond to the thick thing whose plate | board thickness is 1 mm or more.

  According to the present invention, even if there are a plurality of fine shapes and through holes in the plate surface, it can be accurately produced. In addition, a plastic plate having a plate thickness of 1 mm or less can be manufactured by molding only hot embossing. However, the present invention is also a technique that can be used for manufacturing a thick plastic plate having a plate thickness of 1 mm or more. Is useful.

The top view of the biological sample discrimination | determination plate concerning Embodiment 1 of this invention. The top view which expands and shows the pattern of the same biological sample discrimination plate Sectional drawing of sample injection part and buffer injection part of the same biological sample discrimination plate Sectional drawing of sample injection part and buffer injection part of the same biological sample discrimination plate Sectional drawing of sample injection part and buffer injection part of the same biological sample discrimination plate The top view which shows the pattern of the same biological sample discrimination | determination plate The top view which expands and shows the fixed quantity fractionation part of the same biological sample discrimination plate The top view which shows the pattern of the same biological sample discrimination | determination plate The top view which shows the pattern part of the same biological sample discrimination plate The top view which shows the pattern of the same biological sample discrimination | determination plate The top view which expands and shows the fixed quantity fractionation part of the same biological sample discrimination plate The graph which shows the relationship of the curvature of the plate 1a when the plate | board thickness of the plate 1a concerning Embodiment 1 of this invention is changed.

Explanation of symbols

1, 1a, 1b Plate 2 Flow path pattern 3 Rotating part fixing hole 4 Positioning hole 5 Plate center of gravity 6 Buffer injection port 7 Buffer injection port 8 Sample injection port 9 Sample injection port 10 Fixed quantity sorting unit 11, 12 Flow channel 13,15 Air hole 14,16 Overflow chamber 17 Bonding rib 18 Adhesive 21 Film 30 DNA conjugate 31 DNA sample A Electrophoresis direction

Claims (9)

In a biological sample discriminating plate in which a groove and a non-through hole or a through hole are formed and the plate thickness is 1 mm or more, the groove and the through hole are simultaneously formed by hot embossing using a mold. Biological sample discrimination formed by joining the plate 1 and the plate 2 formed with a non-through hole or a through hole by any one of injection molding, injection molding, and machining by an adhesive or thermocompression bonding Plate. The biological sample discrimination plate according to claim 1, wherein the biological sample discrimination plate is a biological sample discrimination plate for discriminating a biological sample. Of the surfaces constituting the biological sample discrimination plate, the groove formed in the first surface is the first surface on which the groove and the non-through hole or the through hole are formed. A surface area occupied on the first surface, a surface area occupied by the non-through holes formed on the first surface on the first surface, and a surface area occupied by the through holes formed on the first surface on the first surface. The biological sample discriminating plate according to claim 1, wherein the total of the total surface area is 5% or more of the total surface area of the first surface. The biological sample discrimination plate according to any one of claims 1 to 3, wherein a thickness of the plate 1 is 0.3 mm or more and 1 mm or less. In the biological sample discriminating plate, a convex portion continuous with a wall surface forming the non-through hole or the through hole is formed on a surface of the plate 2 to be joined to the plate 1. The biological sample discriminating plate according to claim 1. The biological sample discriminating plate according to claim 5, wherein the convex portion has a cylindrical shape having an inner diameter equal to an inner diameter of the through hole of the plate 2. The biological sample discrimination according to claim 5, wherein the convex portion is located at an equal distance from the central axis of the through hole of the plate 1 and is formed so as to be accommodated in the through hole of the plate 1. Plate. 2. The biological sample discrimination according to claim 1, wherein the plate 1 is formed by forming a groove only on one surface and bonding the surface of the plate 1 on which the groove is not formed and the plate 2. plate. In the biological sample discriminating plate in which a groove and a non-through hole or a through hole are formed and the plate thickness is 1 mm or more, the groove and the through hole are simultaneously formed by hot embossing using a mold. The plate 1 and the plate 2 in which a non-through hole or a through hole is formed and a convex portion is formed on one surface are bonded by an adhesive by any of injection molding, injection molding, and machining. In the biological sample discriminating plate manufacturing method for manufacturing the biological sample discriminating plate formed in this manner, the adhesive is applied to the surface of the plate 1 opposite to the surface on which the groove and the through hole are formed, and the plate 1 And a plate manufacturing method for biological sample discrimination, comprising the step of joining the plate 2 and the plate 2 with an adhesive.

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