JP4552548B2 - Sequential transfer reaction tank, manufacturing method of sequential transfer reaction tank, and test method using sequential transfer reaction tank - Google Patents

Description

  The present invention includes, for example, a sequential transfer reaction tank used for confirmation of PCR (polymerase chain reaction method) products, detection by antigen-antibody reaction, measurement of enzyme activity, etc., a method for manufacturing a sequential transfer reaction tank, and a sequential transfer reaction tank. It relates to the test method used.

In recent years, attention has been paid to the fact that a reaction is effectively performed by using a very small amount of sample in a chemical reaction or the like, and an inspection apparatus using such a small amount of sample has been proposed. For example, the substrate is finely processed using a micromachining method, a sample storage unit for storing the sample, a sample storage unit for storing the reagent, a reaction unit for reacting the sample and the reagent, and A flow path connecting these is formed, and various analyzes are performed with a small amount of sample and reagent (see, for example, Patent Document 1). In the analysis, a small amount of sample or reagent is accommodated in each of the accommodating portions, the sample and the reagent are guided to the reaction portion using electric induction, and reacted there.
Japanese Patent No. 3207424

  The above prior art is advantageous in that it requires only a small amount of expensive reagents and valuable samples, but has a problem that it is difficult to manufacture because the substrate must be finely processed. In addition, since the sample and the reagent are moved by using electric induction, the adjustment must be performed according to the viscosity of each sample, and there is a problem that it takes time and labor for the work. Furthermore, since the amount of the sample is very small, there is a possibility that the sample may slightly evaporate when exposed to the outside.

  The present invention has been made in consideration of such circumstances, and does not require advanced processing techniques, can be manufactured easily and has a simple configuration, and can perform analysis quickly and effectively with a small amount of sample. It is an object of the present invention to provide a sequential transfer reaction tank, a method for manufacturing a sequential transfer reaction tank, and a test method using the sequential transfer reaction tank.

The present invention employs the following means in order to solve the above problems. The invention according to claim 1 is a sealed housing portion (a housing portion according to an embodiment of the present invention) for reacting a sample (sample 7 according to an embodiment of the present invention) and a reagent (reagent 6 according to an embodiment of the present invention). S) and a base (base 2 in the embodiment of the present invention) on which the storage part is arranged, and the storage part corresponds to the reagent to be filled, and the partition means (the heat seal 4 of the embodiment of the present invention). , Roll 12, gap c), and is divided into a plurality of small rooms (small rooms 8, 9, 10 of the embodiment of the present invention) so as to be divided and combined.
With this configuration, it is possible to accommodate samples and reagents in a plurality of small rooms as necessary, and to release the partitioning means during the reaction so that the small rooms can be combined to allow the sample and the reagent to react. .

The invention described in claim 2 is characterized in that the accommodating portion is formed in a tube (tube 3 of the embodiment of the present invention).
By configuring in this way, the container is sealed from the outside by being sealed in the tube, and it is possible to prevent the reagent and the sample from evaporating and to reliably block the reagent and the sample from the external environment. Become.

  The invention according to claim 3 is characterized in that the accommodating portion is formed between the base and a film (film 14 of the embodiment of the present invention) covering the upper surface of the base (the upper surface 2a of the embodiment of the present invention). And

The invention described in claim 4 is characterized in that a recess (recess 13 in the embodiment of the present invention) for arranging the reagent or the sample and the reagent is formed on the base.
With this configuration, it is possible to reliably arrange the reagent or the sample and the reagent in the recess.

The invention according to claim 5 is characterized in that the partitioning means is an adhesive portion (heat seal 4 of the embodiment of the present invention) for partitioning the inside of the tube or the film by bonding to the base.
By comprising in this way, it becomes possible to partition the inside of a tube freely with an adhesion part.

The invention described in claim 6 is characterized in that the partitioning means is a pressing tool (roll 12 in the embodiment of the present invention) for partitioning the inside of the tube or the film by crimping to the base.
By comprising in this way, the required small room can be united by opening the pressing tool.

The invention according to claim 7 is characterized in that the partitioning means is a partition part provided with a fine gap (gap c in the embodiment of the present invention) that can be circulated by being pressurized.
By comprising in this way, it becomes possible to give the partition part itself the function to partition an adjacent small room, and the function to unite a small room.

The invention described in claim 8 is characterized in that it is provided with a plurality of sealed small chambers that contain a sample and a reagent that are partitioned by the partitioning means and can be mixed by being pressurized.
By configuring in this way, a plurality of small chambers can contain samples and reagents as necessary, and in the reaction, the partitioning means are opened, the small chambers are combined, and the sample and the reagent are reacted by pressurization. Is possible.

  The invention according to claim 9 includes a step of adhering the tube to the base, a step of putting the reagent or the sample and the reagent in the tube, a step of sealing by the partitioning means, and a step of sealing the open end of the tube. It is characterized by.

  The invention according to claim 10 includes a step of arranging a reagent or a sample and a reagent on a base, a step of covering the base with the reagent or the sample and the reagent with a film, a step of sealing with a partitioning means, a film and the base And the step of closing the open end between them.

  The invention according to claim 11 is a step of filling at least one sealed small chamber with a sample, and a step of opening a compartment between the small chamber filled with the sample and a small chamber filled with an adjacent reagent; The method comprises the steps of pressurizing the small chamber filled with the sample and mixing it with the reagent in the adjacent reagent small chamber, and stirring the mixture.

  According to the first aspect of the present invention, it is possible to accommodate samples and reagents in a plurality of small rooms as required, and to release the partitioning means when the reaction is performed so that the small rooms are combined and the sample and the reagent can be reacted. Therefore, there is an effect that the manufacturing is easy and simple, and the analysis can be performed quickly and effectively with a small amount of sample.

  According to the second and third aspects of the present invention, the container is sealed from the outside by being sealed in the tube or film, so that the reagent and the sample are prevented from evaporating and the reagent and the sample are surely removed from the external environment. Since it is possible to block, there is an effect that the experiment accuracy can be improved by eliminating the variation in the experiment result due to the concentration change of the valuable reagent, the sample, and the contamination.

  According to the fourth aspect of the present invention, it is possible to reliably arrange the reagent or the sample and the reagent in the concave portion by providing the concave portion in the base. effective.

  According to the invention described in claim 5, since it becomes possible to freely partition the inside of the tube or the film by the adhesive portion, there is an effect that the degree of freedom of partitioning the small room can be increased according to the amount of the sample or the reagent. is there.

  According to the invention described in claim 6, since the necessary small rooms can be united by opening the pressing tool, there is an effect that damage to the tube or the film can be minimized.

  According to the invention of claim 7, since it becomes possible to have the function of partitioning adjacent small rooms and the function of combining small rooms in the partition part itself, once the partition part is formed, Since there is no need to open the partition part later, there is an effect that the experiment process can be reduced.

  According to the invention described in claim 8, samples and reagents are accommodated in a plurality of small rooms as required, and in the reaction, the partitioning means are opened to combine the small rooms, and the sample and the reagent are reacted by pressurization. Therefore, there is an effect that the manufacturing is easy and simple, and the analysis can be performed quickly and effectively with a small amount of sample.

  According to the invention described in claim 9, it is possible to manufacture a sequential transfer reaction tank easily and reliably at low cost without requiring fine processing.

  According to the invention described in claim 10, since the reagent or the sample and the reagent are arranged on the base before covering with the film, there is an effect that the injection of the reagent becomes easy.

  According to the eleventh aspect of the invention, since the sample and the reagent are moved, mixed, and stirred by using the united compartments, the sample is not affected by the viscosity of each sample. It is possible to reliably mix the two, so that the test can be easily performed in a short time. Furthermore, since the sample in the sealed small room does not evaporate, there is an effect that it is easy to handle.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
FIG. 1 is a diagram showing a first embodiment of the present invention, and shows a sequential transfer reaction tank to which the present invention is applied.
The sequential transfer type reaction tank (hereinafter referred to as a chip) 1 has a generally cylindrical tube 3 on a substantially rectangular plate-like substrate 2, an adhesive 4 and a thin plate-like heat seal 4. And partitioning means such as are provided. Hereinafter, the case where the heat seal 4 is used as partitioning means will be described.

  The substrate 2 is made of a glass substrate, a metal plate, PC (polycarbonate), PDMS (polydimethylsiloxane), PMMA (methacrylic resin), etc. The vertical and horizontal dimensions of the tube 3 can be arranged, and at most 10 cm in length and width. It is formed to have a thickness that does not easily bend during work. And the said tube 3 is adhere | attached and provided in the base | substrate upper surface 2a so that it may become parallel to the longitudinal direction of the base | substrate 2. As shown in FIG.

  The tube 3 is formed of a flexible material in a cylindrical shape, and is configured as a housing portion S whose inside is sealed. For example, PP (polypropylene), PET (polyethylene terephthalate resin) , Ny (nylon), PC (polycarbonate), PVC (polyvinyl chloride), PVDC (polyvinylidene chloride) or other single-layer film or multilayer film for containing reagent 6 and sample 7 . The diameter of the tube 3 is several tens of μm to several cm, and the length thereof can be freely adjusted according to the reaction mode. Further, the thickness dimension is such that the reagent 6 or the sample 7 is provided inside, and the thickness does not break even when mechanically pressurized or finger pressed from the outside.

And either one edge part of the tube 3 is sealed, the check valve 5 is provided, and the other edge part is sealed. Here, the heat seal 4 (the first heat seal 4a and the second heat seal 4b) is welded (indicated by an arrow) with a weak adhesive force that is opened when the tube 3 is pressed, and reliably. It has become.
Instead of providing the check valve 5, a hole may be formed in the tube 3, the reagent 6 or the sample 7 may be injected by the syringe 17, and sealed with the heat seal 4.

Next, the operation of the first embodiment will be described.
As shown in FIG. 1, the small chamber 8 of the chip 1 is filled with the sample 7, the small chamber 9 is filled with the reagent 6, and the small chamber 10 is filled with another reagent 6, as shown in FIG. When pressure is applied (indicated by an arrow), the first heat seal 4a (indicated by a chain line) is opened, and the sample 7 in the small chamber 8 is mixed with the reagent 6 in the small chamber 9. Next, if any one of the small chamber 8 and the small chamber 9 is further pressed intermittently, the reagent 6 and the sample 7 are stirred inside the unified small chambers 8 and 9. In this state, the reaction is observed as necessary. Further, when pressure is applied to both of the small chambers 8 and 9, the second heat seal 4b is opened, and the small chamber 10 is united with them to become a unified chamber. Are mixed. Then, if any part of the small chamber 8 to the small chamber 10 which is integrated as a whole is intermittently pressed, another reagent 6 and the sample 7 are formed inside the unified small chamber 8 to small chamber 10. Stir. In this state, the reaction by this other reagent 6 is observed.

  According to the first embodiment, the first heat seal 4a and the second heat can be easily obtained by storing the sample 7 in the plurality of small rooms 8 and different reagents 6 in the small rooms 9 and 10 and applying pressure. Since the seal 4b is opened and the small chamber 8 to the small chamber 10 are combined as necessary, and the reagent 6 and the sample 7 can be reacted, the manufacturing is easy and simple, The sample 7 can be analyzed quickly and effectively. Therefore, adjustment work is not required without being affected by the viscosity of each reagent 6 and sample 7.

  In addition, since the storage portion S is sealed in the tube 3, it is covered from the outside, and it is possible to prevent the reagent 6 and the sample 7 from evaporating and to reliably block the reagent 6 and the sample 7 from the external environment. Therefore, the variation in the concentration of the valuable reagent 6 and the sample 7 and the variation in the experimental result due to the mixing of foreign matters can be eliminated, and the experimental accuracy can be improved.

  Here, since the positions of the first heat seal 4a and the second heat seal 4b can be freely set, the degree of freedom of partitioning of the small rooms 8 to 10 is set according to the amount of the reagent 6 and the sample 7. Can be increased.

  In this way, the reagent 6 and the sample 7 are moved and mixed by using the united compartments 8 to 10, and therefore, affected by the viscosity of each reagent 6 and the like. The test time can be shortened by reliably mixing the two, and the test in the sealed small chambers 8 to 10 prevents evaporation of the reagent 6 and facilitates handling. Can be achieved.

Next, a manufacturing method of the chip 1 of the first embodiment will be described.
First, as shown by the arrows in FIG. 3, a portion of the tube 3 is welded to the upper surface 2 a of the base 2 by ultrasonic waves or heat to form a heat seal 40 having a strong adhesive force, and the tube 3 is fixed. Alternatively, the tube 3 is fixed to the upper surface 2a of the base 2 using an adhesive.
Next, as shown in FIG. 4, a first heat seal 4a is formed on the side where the check valve 5 is located by ultrasonic waves or heat in a direction transverse to the width direction of the tube 3, and a small chamber 8 is formed there. Are sealed. Then, as indicated by the arrow, the reagent 6 is inserted from the open end side of the tube 3, and the second heat seal 4b is formed again at a predetermined interval to seal the small chamber 9. Here, the first heat seal 4a and the second heat seal 4b have a weak adhesive force.
Next, another reagent 6 is put between the second heat seal 4 b and the open end of the tube 3, and the open end of the tube 3 is closed with the heat seal 4. In using the chip 1, the sample 7 may be injected into the small chamber 8 from the check valve 5.
Therefore, according to this manufacturing method, the chip 1 can be manufactured easily and reliably at low cost without requiring fine processing.
Here, it is also possible to adopt a method in which a small chamber is formed in advance prior to the introduction of the reagent 6 and the like, a hole is formed, the reagent 6 is injected into each small chamber, and the open end of the hole and the tube 3 is closed.

Next, a second embodiment of the present invention will be described with reference to FIGS. The same components as those in the first embodiment are denoted by the same reference numerals, description thereof is omitted, and differences will be mainly described.
The chip 1 ′ of the second embodiment uses, for example, a metal rod or a roll 12 (partitioning means) instead of the heat seal 4 of the first embodiment. Specifically, the tube 3 is pressed against the base 2 by the first roll 12a, the inside of the tube 3 is crimped, the small chamber 8 and the small chamber 9 are partitioned, and the tube 3 is similarly formed by the second roll 12b. Is pressed against the base 2 to crimp the inside of the tube 3 to partition the small chamber 9 and the small chamber 10. On the other hand, if the pressing of any one of the rolls 12 is released, the adjacent small chambers are united and the reagent 6 in each small chamber is mixed. Moreover, each roll 12 can also perform stirring operation etc. simultaneously by moving along the longitudinal direction of the tube 3 in the state which spaced apart from the upper surface 2a of the base | substrate 2 a little upwards, and pressed the tube 3. As shown in FIG.
Therefore, according to the second embodiment, since the necessary small rooms can be combined by opening the roll 12, damage to the tube 3 can be minimized.

Next, a third embodiment of the present invention will be described with reference to FIGS. The same components as those in the first embodiment are denoted by the same reference numerals, description thereof is omitted, and differences will be mainly described.
The chip 20 according to the third embodiment includes a base 2 and a film 14 that covers the upper surface 2 a of the base 2. Specifically, like the tube 3, it consists of a single layer film of the above-mentioned material or a multilayer film. And the diameter of the film 14 is several 10 micrometers-several cm, The length shall be freely adjustable with the reaction form.
Further, the periphery of the film 14 is welded around the upper surface a of the substrate 2 by a heat seal 14 a, and a sealed housing portion S is formed between the upper surface 2 a of the substrate 2 and the film 14.
Alternatively, the sealed housing portion S is formed by bonding the upper surface 2a of the base 2 and the film 14 using an adhesive.
The upper surface 2a of the base 2 is formed with a concave portion 13 for arranging the reagent 6 and the like, and the concave portion 13 has a shape in which the reagent 6 such as solid and liquid can be arranged. In this embodiment, three recesses 13 are formed. A check valve 5 is provided at one end of the accommodating portion S. And the heat seal 4 is provided so that it may partition with each recessed part 13 of the base | substrate 2, and several small chambers (for example, three) 8, 9, and 10 are dividedly formed in the accommodating part S. FIG.
According to the third embodiment, since the solid and liquid reagents 6 can be reliably disposed in the recesses 13, the reagent 6 can be easily accommodated in the small chambers.

Next, a manufacturing method according to the third embodiment will be described. As shown in FIGS. 7 to 9, the chip 20 is formed with a plurality of recesses 13 (three in this case) on the upper surface 2a of the base 2 by machining, and each of the recesses 13 is filled with each reagent 6 or the like. Then, the peripheral edge of the film 14 is sealed with a heat seal 14 a so as to cover the reagent 6 and the substrate 2, thereby forming the accommodating portion S. Then, the base 2 between the inside of the film 14 and each recessed part 13 is heat-sealed using the heat seal 4, and each small room 8-the small room 10 are partition-formed.
According to the manufacturing method of the third embodiment, the chip 20 can be manufactured easily and reliably at low cost without requiring fine processing.

Next, a fourth embodiment of the present invention will be described with reference to FIG. The same components as those in the first embodiment are denoted by the same reference numerals, description thereof is omitted, and differences will be mainly described.
The heat seal 4 of the first embodiment is formed over the entire width in the direction transverse to the width direction of the tube 3, whereas the tip 30 of the fourth embodiment is pressurized. A first heat seal 4c and a second heat seal 4d are formed, leaving a fine gap c (partitioning means) that can be circulated. Therefore, in the normal state where no pressure is applied by the first heat seal 4c and the second heat seal 4d, the small chambers 8 to 10 are partitioned, and when pressure is applied, the reagent is used between the small chambers by the gap c. 6 etc. can be distributed.
Therefore, according to the fourth embodiment, the first heat seal 4c and the second heat seal 4d themselves can have a function of partitioning adjacent small rooms and a function of combining the small rooms. Once the first heat seal 4c and the second heat seal 4d are formed, it is not necessary to open the first heat seal 4c and the second heat seal 4d later, so that the test process can be reduced.

A case where the above-described chip is actually used for the test will be described with reference to FIGS.
In an actual test, the chip 1 is set and used in an inspection apparatus 18 to be described later, but in the following description, from injection to detection of a sample will be briefly described. Here, the sample 7 is blood, an extracted sample, a PCR product, or the like.
First, confirmation of the polymerase chain reaction (PCR) product will be described.
As shown in FIG. 11, the sample 7 is injected into the first small chamber 8 from the check valve 5 using the syringe 17. For convenience of illustration, only the reference sign is shown for the reverse valve 5 and the illustration is omitted. Next, after the injection is completed, by applying pressure as indicated by an arrow in FIG. 12, the heat seal 4 that has been weakly bonded is opened as indicated by a broken line, and the first small chamber that has been partitioned. 8 and the second small room 9 are unified. It is assumed that the second chamber 9 of the tube 3 is filled with, for example, a freeze-dried polymerase chain reaction (PCR) reagent or a liquid reagent. At this time, when stirring is necessary, stirring is performed by ultrasonic waves, vibration, rotation, and a roll 12 described later. Next, as shown in FIG. 13, the whole is heated and cooled by a heat block, and a polymerase chain reaction (PCR) is performed. After completion of the reaction, as shown in FIGS. 14 and 15, the pressure is applied again to unify with the third chamber (detection unit) 10 filled with the fluorescent reagent, and the PCR reaction solution and the detection solution are mixed. The PCR product is confirmed by fluorescence detection.
The detection method is not limited to this, and colorimetry, chemiluminescence, precipitation, heat generation, RI method and the like can also be used.

Next, antigen detection by antibody antigen reaction will be described. Since the substantial procedure is the same as the confirmation of the polymerase chain reaction (PCR) product, it will be described with reference to FIGS.
First, a chip 1 is prepared in which a tube having a length of 25 mm and a diameter of 5 mm is set on a rectangular base of 30 mm × 70 mm. The tube is partitioned by using a heat seal 4 so that the lengths of the first small chamber 8, the second small chamber 9, and the third small chamber 10 are 5 mm, 10 mm, and 10 mm, respectively. The second small chamber 9 of the chip 1 is filled with a DNA degrading enzyme and a surfactant, and the third small chamber 10 is filled with an antibody against the antigen to be detected. Here, for example, the DNA-degrading enzyme and the surfactant are 50 microliters in total, specifically, 20 mM Tris-HCl (PH7.4), 2 mM NaCl (sodium chloride), 10 mM EDTA, 2% Triton X It consists of −100, 2% DOC, 0.2% SDS, and 2% Traylol.
As shown in FIG. 11, the sample 7 is injected into the first small chamber 8 from the check valve 5 using the syringe 17. Next, after the injection is completed, by applying pressure as indicated by an arrow in FIG. 12, the heat seal 4 that has been weakly bonded is opened as indicated by a broken line, and the first small chamber that has been partitioned. After unifying 8 and the second small chamber 9, as shown in FIG. 13 by any method such as ultrasonic wave, vibration, rotation and roll 12 described later (in this example, heat block is not performed) ) Stir. After the agitation, as described above, the heat seal 4 interposed between the third small chamber 10 (detection unit) by applying pressure again to the unified first small chamber and second small chamber 9. As shown by the broken line in FIG. 14, the reaction solution is moved from the opened compartment to be unified with the third chamber 10 (detection unit), and the antigen-antibody reaction is performed in FIG. Here, if aggregation occurs, it can be determined that the target antigen is contained in the sample. Therefore, antigen detection is performed by a precipitation method.

Next, enzyme activity measurement will be described with reference to FIGS.
The second small chamber 9 of the chip 1 is filled with a DNA degrading enzyme and a surfactant. Then, the third chamber 10 (detection unit) is filled with a fluorescent reagent that develops a color by separating the quencher fluorescent dye by protease if it is a peptide labeled with a fluorescent dye and a quencher by an enzymatic reaction. Yes.
Also in this case, the sample 7 is injected from the check valve 5 of the first small chamber 8 using the syringe 17 (FIG. 11). After the injection is completed, by applying pressure as shown by the arrow in FIG. 12, the weakly bonded heat seal 4 is opened as shown by the broken line, and the reagent 6 and the sample 7 are removed from the opened compartment. Move (FIG. 13). After unifying the first small chamber 8 and the second small chamber 9, stirring is performed by any method such as ultrasonic waves, vibration, rotation, and a roll 12 described later. At this time, dissolution of the sample 7 is also included. After the stirring and dissolution of the sample 7 are completed, as described above, pressure is applied again to open the heat seal 4 (FIG. 14). Then, the reaction solution is moved in a unified manner with the third small chamber 10 (detection unit) (FIG. 15), and a protease that cleaves the labeled peptide or protein is measured.
Various enzyme activity measurements can be performed by combining the enzyme and the substrate.

Next, an inspection apparatus (hereinafter referred to as a kit) 18 on which the chip 1 is set will be briefly described with reference to FIG. In the figure, the kit 18 includes a mounting part A for mounting the chip 1, a filling part B for filling the reagent 6 and the sample 7, a first pressurizing part C, a heating part D, and a second pressurizing part E. The display unit F is provided.
The chip 1 filled with the reagent 6 and partitioned into the small chambers is first set in the mounting portion A. And the sample 7 is filled from the filling part B, moves to the 1st pressurization part C, applies a pressure to a 1st small room, and unifies with a 2nd small room. After that, for the inspection that requires heating, the whole is heated by moving to the heating part D. If stirring is necessary at this time, stirring is performed simultaneously. Further, the second pressurizing unit E applies pressure to the unified first small room and the second small room so as to be unified with the third small room and observes on the display unit F.
In addition, the kit 18 may be provided with a roll 12 in some cases. After the chip 1 is mounted on the mounting portion A of the kit 18 without being filled with the reagent 6 and partitioned, the section is partitioned by pressing the roll 12. It is formed. Then, the reagent 6 and the sample 7 are filled into the small chambers from the filling part B through the check valve 5 provided in the tube 3. Thereafter, the adjacent small chambers are unified by moving the roll 12, and at the same time, the reagent 6 and the sample 7 are stirred.
Here, the chip 1 moving in the kit 18 is not performed in the order described above, and can be moved to any part according to the contents of inspection. The kit 18 may be configured to form the heat seal 4 of the chip 1.

The present invention is not limited to the above-described embodiment, and the following various aspects can be employed. The upper surface 2a of the base 2 does not necessarily need to be smooth, and may be formed by providing a recess according to the shape of the tube 3 in order to stabilize the tube 3 on the base 2, and the shape of the tube 3 is cylindrical. However, it is not limited to this. In addition, the small room (here, the third small room 10) in which the final reaction is performed, that is, not only the detection unit, but any small room can be changed in size depending on the reaction form. The movement of the sample to the detection unit may be performed by evacuation instead of being pressurized. Or it is also possible to make only the last detection part into a pressure reduction state previously. The position of the last detection unit is not determined, and may be, for example, a central small room. Further, a plurality of detection units may be provided according to the number of reagents 6 to be measured.
Further, the chip 1 can be used in a vertically standing state, or may be used only when the reagent 6 and the sample 7 are moved.

It is a perspective view which shows the chip | tip of 1st Embodiment in this invention. It is a perspective view explaining the effect | action of 1st Embodiment in this invention. It is a side view which shows the manufacturing method of the chip | tip of 1st Embodiment in this invention. It is a side view which shows the manufacturing method of the chip | tip of 1st Embodiment in this invention. It is a perspective view which shows the chip | tip of 2nd Embodiment in this invention. It is a perspective view explaining the effect | action of 2nd Embodiment in this invention. It is a perspective view which shows the chip | tip of 3rd Embodiment in this invention. It is explanatory drawing which shows the manufacturing method of 3rd Embodiment in this invention. It is explanatory drawing which shows the manufacturing method of 3rd Embodiment in this invention. It is a perspective view which shows the chip | tip of 4th Embodiment in this invention. It is explanatory drawing which shows the test example using the chip | tip of embodiment of this invention. It is explanatory drawing which shows the test example using the chip | tip of embodiment of this invention. It is explanatory drawing which shows the test example using the chip | tip of embodiment of this invention. It is explanatory drawing which shows the test example using the chip | tip of embodiment of this invention. It is explanatory drawing which shows the test example using the chip | tip of embodiment of this invention. It is a schematic structure figure of the kit which mounts the chip of the embodiment of the present invention.

Explanation of symbols

2 Base 2a Top 3 Tube 4 Heat seal (partitioning means)
6 Reagent 7 Sample 8, 9, 10 Small room S Container

Claims (11)

  A sealed container for reacting a sample and a reagent, and a base on which the container is arranged, and the container is divided into a plurality of small rooms by partition means corresponding to the sample and the reagent to be filled. And a sequential transfer reaction tank characterized by being partitioned so as to be combined. The sequential transfer reaction tank according to claim 1, wherein the container is formed in a tube. The said transfer part is formed between the base | substrate and the film which covers this base | substrate upper surface, The sequential transfer reaction tank of Claim 1 characterized by the above- mentioned. 4. The sequential transfer reaction tank according to claim 3, wherein a recess for arranging the reagent or the sample and the reagent is formed on the base. It said partitioning means is sequentially transferred type reactor according to claim 2, wherein or 3, characterized in that an adhesive portion for partitioning by adhering the tube or the film. Said partitioning means is sequentially transferred type reactor according to claim 2, wherein or 3, characterized in that said tube or said film is a pusher for partitioning and pressed into the base. The sequential transfer reaction tank according to any one of claims 1 to 4, wherein the partition means is a partition portion having a fine gap that can be circulated by being pressurized. Partitioned by the partitioning means, to any of claims 1 to 7, characterized in that it comprises a plurality of sealed the small chamber for accommodating the reagent or sample and reagent can be mixed by being pressed The sequential transfer reaction tank as described . A method for producing a sequential transfer reaction layer according to any one of claims 1, 2, 5-8,
Bonding the tube to the substrate;
A step of placing the reagent or sample and reagent into the tube,
Sealing with partition means;
Sequential manufacturing method of the transfer type reaction vessel, characterized in that and a step of closing the open end of the tube. A method for producing a sequential transfer reaction layer according to any one of claims 1 to 3,
Placing a reagent or sample and reagent on a substrate;
A step of covering said reagent or sample and reagent and the base in the film,
Sealing with partition means;
Sequential manufacturing method of the transfer type reaction vessel, characterized in that and a step of closing the open end between said base and said film. A test method using the sequential transfer reactor according to any one of claims 1 to 8,
Filling the sample into at least one sealed chamber;
A step of reagent adjacent to the small chamber in which the sample is filled to open the compartments of the small chamber filled,
Test methods successively with transfer type reaction vessel, characterized in that and a step of mixing and stirring with the reagent in the small room reagent adjacent pressurizes small chamber in which the sample has been filled.

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