WO2007122819A1 - Device for reactions with liquid media - Google Patents

Abstract

A biochemical reaction device capable of realizing speedy biochemical reaction, which is provided with reagent reservoir units (10) for feeding reagents to be used either in a solid-liquid reaction or in pretreatment thereof, a sample feed unit (20) for feeding a sample, concentration units (40) for preparing or preserving solutions for the solid-liquid reaction or pretreatment thereof wherein the solutions can be concentrated or evaporated to dryness by distilling away the solvents of the solutions, a reaction unit (70) for conducting the solid-liquid reaction, a passage system (60) for connecting the above units in such a way that the solid-liquid reaction can be carried out, and a fluid driving unit (200) for making a liquid or gas for the solid-liquid reaction flow through the passage system (60).

Description

 Specification

 Equipment for liquid-based reactions

 Technical field

 The present invention relates to an apparatus for performing a reaction using a liquid as a medium.

 Background art

 [0002] In the medical field, diagnoses the severity of diseases and symptoms using information on the presence and concentration of various compounds obtained by analyzing samples such as blood and urine collected from patients as indicators. Things have been done. In recent years, there has been a demand for promotion of personalized medicine based on individual genetic information from the viewpoints of public health promotion and medical economy. In other words, disease, constitution, drug sensitivity, treatment prognosis, etc. are diagnosed by gene polymorphism analysis, chromosome analysis, expression profiling analysis, and proteome analysis. There is a growing need for treatment. According to personalized medicine, overdose of drugs is reduced, side effects on patients are reduced, and people's health is improved, resulting in reduction of medical costs. In addition, preventive medical care can be promoted by making it possible to diagnose an individual's constitution.

 [0003] In order to obtain individual biochemical information such as genetic information that is the basis of diagnosis, techniques such as PCR, nucleic acid hybridization, and antigen-antibody reaction are used. For example, nucleic acid hybridization using a DNA microarray generally requires operations such as extraction of DNA from a sample, introduction of a fluorescent reagent, and hybridization and washing on a DNA microarray (DNA Microarray combat manual, Yotei, published in January 2000. For this reason, these various types of biochemical information are usually performed manually by skilled engineers and using a plurality of large devices. In addition, such analysis usually required about 3 months.

[0004] On the other hand, as a result of the development of microfabrication technology and surface treatment technology for substrates and the like in recent years, liquids are fed, separated, mixed, and reacted on solid phase bodies such as substrates. Development of various reaction devices such as TAS, lab-on-chip, and microchemical chips is now underway.

 Disclosure of the invention

 [0005] In order to realize personalized medical care, various biochemical information can be acquired not only in large-scale medical facilities but also in medical sites such as small and medium-sized medical facilities that exist in a distributed manner. is important. However, it is impossible to carry out an analysis that takes several days until the result is obtained as described above at an individual medical site. In addition, it is difficult to carry out analysis at individual medical sites from the viewpoint of the large size of the equipment and the difficulty of arranging specialist engineers. Furthermore, at present, there is no device that can quickly and easily obtain the above-mentioned biochemical information at a medical site.

 [0006] Accordingly, an object of the present invention is to provide an apparatus capable of quickly acquiring various biochemical information. Another object of the present invention is to provide a compact apparatus for acquiring various biochemical information.

 [0007] In order to solve the problem of quickly obtaining biochemical information, the inventors have made the device compact, and personalized medicine provides information on various types of items for a single specimen. Focusing on the points that need to be acquired and the need to analyze these various types of items simultaneously, at least one step of these reactions is a liquid-based reaction such as a solid-liquid reaction. It was decided to. Then, it was found that by supplying a reaction solution containing a test component in a minute amount and at a high concentration to the reaction process, the diffusion rate limiting of the reaction can be suppressed or eliminated, and the reaction can be accelerated more quickly than expected. The present invention has been completed by finding that the entire system can be speeded up and the apparatus can be made compact. According to the present invention, the following means are provided.

[0008] According to the present invention, there is provided a reaction apparatus for a reaction using a liquid as a medium, the reagent storing unit storing a reagent used for the reaction or its pretreatment, and a sample being introduced A sample introduction part, a concentration part capable of concentrating or drying the solution by distilling off the solvent of the solution while preparing or storing a solution prepared for the reaction or its pretreatment, and the reaction A reaction unit that performs the reaction, a flow channel system that communicates the respective units so that the reaction can be performed, and a fluid drive unit that circulates the liquid or gas for the reaction through the flow channel system. An apparatus is provided. These parts and the channel system are preferably provided in a solid phase body. In these reaction apparatuses, the reaction is preferably a solid-liquid reaction, and is preferably a solid-liquid reaction relating to biomolecules such as nucleic acids and proteins.

 [0009] In the present reaction apparatus, the solution in the concentration unit may be dried in the concentration unit, and the concentration or drying of the solution is accompanied by suction or heating in the concentration unit. It may be. Further, in this reaction apparatus, the concentration unit has a flow direction of the solution in the concentration unit as a longitudinal direction, a direction substantially perpendicular to the flow direction of the solution is a short direction, and the short direction is a solution The width of the supply site and the width of the solution discharge site can be 4 times or less.

[0010] In the present reaction apparatus, the solution in the concentrating unit can be concentrated or dried to a predetermined part of the concentrating unit. In this embodiment, the dry or concentrated part of the solution may be in the vicinity of the connecting part to the reaction rear stage side in the reaction apparatus. Furthermore, the dried or concentrated portion of the solution may be a reagent reservoir portion from the reagent reservoir. The concentration unit has a flow direction of the solution in the concentration unit as a longitudinal direction, a direction substantially perpendicular to the flow direction of the solution as a short direction, and the short direction includes the width of the solution supply site and the above-described direction. The width can be less than or equal to twice the width of the solution discharge site. Furthermore, the concentration part of the solution remaining in the concentration part may be visible from outside the concentration part.

[0011] The present reaction apparatus includes a breathable material that is exposed in the concentration section and has liquid repellency with respect to the reaction solution and can discharge the gas in the concentration section, and the gas in the concentration section is the breathable material described above. And a ventilation part for distilling the solvent out of the concentration part through the air-permeable material. In this embodiment The concentration unit may include the breathable material in at least a part of the lower surface in the gravitational direction where the reaction solution supplied to the concentrating unit comes into contact, and the concentrating unit is provided on the lower surface in the gravitational direction. The breathable material may be provided on almost the entire surface. In addition, the ventilation portion may be provided so that gas in the concentration portion can be sucked from the entire surface of the breathable material exposed in the concentration portion, or a part of the breathable material exposed in the concentration portion. The aeration part may be provided so that the gas in the concentration part can be sucked.

 [0012] Further, the present reaction apparatus comprises a breathable material that is exposed in the concentrating part and has liquid repellency with respect to the reaction liquid and can exhaust the gas in the concentrating part. A vent part that has substantially the entire surface, and when the gas in the concentrated part is sucked through the breathable material, the solvent is distilled out of the concentrated part through the breathable material; The solution in the concentration part can be concentrated or dried to a predetermined part of the material, and the predetermined part can be a reagent storage part from the reagent storage part. Further, the concentration unit has a flow direction of the solution in the concentration unit as a longitudinal direction, a direction substantially perpendicular to the flow direction of the solution as a short direction, and the short direction includes a width of a solution supply site and The width of the solution discharge site is 4 times or less, and the concentration site of the solution in the concentration unit can be made visible from outside the concentration unit.

 [0013] In the present reaction apparatus, the solution can be mixed by forming a differential pressure in the concentration section. Further, in this aspect, the differential pressure is exposed in the concentrating part, and has a liquid repellency with respect to the reaction liquid, and the gas in the concentrating part is discharged through a gas-permeable material capable of discharging the gas in the concentrating part. Can be formed by suction.

[0014] The present reaction apparatus can further include a purification unit for purifying a test component in a solution prepared for the reaction or its pretreatment. Further, in this embodiment, the purification unit may include a monolith column, and the monolith column may be capable of desorbing the test component using water as an eluent after adsorbing the test component. . Furthermore, it contains a test component purified by the purification unit The eluate can be concentrated or dried in the concentration section. In addition, the test component may be a nucleic acid, or the solution or the reagent may be introduced into the monolith column in a pressurized or reduced pressure state.

 [0015] Further, in the present reaction apparatus, the reaction part has a flow direction of the solution in the reaction part as a longitudinal direction, a direction substantially perpendicular to the flow direction of the solution as a short direction, and the short direction. The width of the solution supply site and the width of the solution discharge site may be 4 times or less. In this embodiment, the test component is a nucleic acid, and the reaction in the reaction part can be a nucleic acid hybridization reaction. Further, the nucleic acid hybridization can be an array CGH. The concentration of the nucleic acid that is the test component in the reaction part may be 0.3 g Z I or more.

 Brief Description of Drawings

[0016] FIG. 1 is a diagram showing an outline of an example of a reaction apparatus for carrying out array CGH.

 FIG. 2 is a diagram showing an outline of an example of a concentrating part of a reaction apparatus, in which a plan view is shown in the upper part and a sectional view is shown in the lower part.

 FIG. 3 is a diagram showing an example of a solid-liquid reaction part of a reaction apparatus, in which a plan view is shown in the upper part, and a sectional view and a manufacturing process thereof are shown in the lower part.

 FIG. 4 is a diagram showing an example of the flow of an array C G H using this reaction apparatus.

[FIG. 5] Fluorescence signal ratio I og ₂ (by array CGH in Example 1 and comparative example)

It is a figure which shows the comparison result of Cy3ZCy5).

 FIG. 6 is a view showing a scanned image obtained in Example 1.

 BEST MODE FOR CARRYING OUT THE INVENTION

The present invention relates to a reaction apparatus and reaction method for reaction using a liquid as a medium. The reaction apparatus of the present invention prepares a reagent storage unit for storing a reagent used for the reaction or its pretreatment, a sample introduction unit for introducing a sample, and a solution prepared for the reaction or its pretreatment Alternatively, the concentration part that can concentrate and dry the solution by distilling off the solvent of the solution while being stored, the reaction part that performs the reaction, and the flow that communicates the parts to enable the solid-liquid reaction. Path system and liquid for the reaction A fluid drive unit for circulating a body or gas through the flow path system. According to the reaction apparatus of the present invention, a solution for reaction using a liquid as a medium can be concentrated or dried. For this reason, the liquid for reaction can be prepared by supplying the liquid to the dried product or the concentrate to the reaction solution containing the test component at a high concentration. As a result, the reaction can be performed at a high concentration of the test component. The high concentration of the test component allows the reaction to be carried out quickly by suppressing or avoiding the diffusion rate limiting of the reaction, and biochemical information can be obtained quickly.

 [0018] Further, according to the present reaction apparatus, the reaction can be carried out by supplying a liquid necessary for the reaction or the like to the dried or concentrated product, so that the amount of the liquid for the reaction can be suppressed. As a result, the reaction can be further accelerated and the reaction section can be implemented in a compact manner.

 [0019] The concentration unit can also be provided for preparing or storing a solution prepared for the pretreatment of the reaction. Since these solutions (pretreatment liquids) are concentrated or dried, the liquid required for the next step can be supplied to prepare a pretreatment solution containing the test component at a high concentration. . For this reason, various pretreatment steps can be carried out in a state where the concentration of the test component is high and the amount of the liquid is suppressed, and as a result, various pretreatments leading to reactions using the target liquid as a medium can be performed quickly. And it can be implemented in a compact manner.

 Hereinafter, a reaction apparatus and a reaction method which are embodiments of the present invention will be described with reference to the drawings as appropriate. FIG. 1 to FIG. 4 relate to a reaction apparatus for performing array CGH, which is a solid-liquid reaction by extracting DNA from blood, and FIG. 1 is a diagram showing an outline of an example of the reaction apparatus of the present invention. FIG. 2 is a diagram showing an outline of an example of the concentration unit of the reaction apparatus of the present invention, FIG. 3 is a diagram showing an example of the reaction unit of the reaction apparatus of the present invention, and FIG. 4 uses the present reaction apparatus. It is a figure which shows an example of the flow of the reaction process which had been. Note that the reaction apparatus and a part thereof shown in these drawings are only examples for explaining the present invention, and do not limit the present invention.

[0021] The present reaction apparatus is an apparatus for carrying out a reaction using a liquid as a medium. liquid There is no particular limitation on the reaction using a medium. Reactions at the interface such as liquid-liquid reaction, gas-liquid reaction, solid-liquid reaction, etc. are also included in “reaction using liquid as medium” in this reaction apparatus. The present reactor 2 is preferably one that performs a solid-liquid reaction. Solid-liquid reactions are diffusion-controlled and generally take a considerable amount of time. The “reaction” in the present invention includes not only general chemical reactions and electrochemical reactions such as synthesis and decomposition, but also various molecular interactions such as hydrogen bonds, hydrophobic bonds, dipole interactions, and van der Waals forces. Includes expression or change. For example, reactions that can be performed as solid-liquid reactions include antigen-antibody reactions; various enzyme reactions; detection of specific genes and SNPs, array CGH, expression profiling, and other nucleic acid hybridization; PCR; Examples include nucleic acid synthesis reactions such as DNA.

[0022] The reaction apparatus is an apparatus for carrying out a reaction using a liquid as a medium. In carrying out the reaction, another reaction can be carried out. For example, to prepare a solution suitable for the reaction, dissolution, dilution, concentration, mixing, separation, extraction, washing, adsorption, desorption, drying, heating, cooling, irradiation with various electron beams, ultrasonic treatment and Examples include pretreatment reactions that include elements such as electrical treatment. For example, in order to perform nucleic acid hybridization between a biological specimen and a probe prepared in advance, separation, purification, fragmentation, and labeling of DNA from the specimen are required as pretreatment.

 [0023] The liquid used as a reaction medium is not particularly limited. In addition to water, various organic solvents and mixtures thereof are included. These liquids can also contain solutes as a part of a medium such as a salt such as a buffer solution. In the reaction apparatus of the present invention, when the antigen-antibody reaction, the enzyme reaction, the nucleic acid hybridization, and the cell-related reaction are performed, the liquid is often an aqueous liquid containing water.

[0024] The component (reaction component) involved in these reactions can include cells in addition to compounds. The compounds can include DNA, RNA, DNAZ RNA cameras, or nucleic acids including these derivatives, proteins, saccharides, and the like. Nucleic acids and proteins are pre-labeled with fluorescent dyes. May be. Examples of proteins include enzymes, antibodies, antigens, ligands and receptors, and peptides. In the case where the reaction component is a cell, the reaction includes expression and change of interaction between cells, cell culture, cell destruction (or extraction associated therewith), cell separation, and the like. The cells mentioned here include microorganisms (bacteria, fungi, viruses, etc.), animal and plant cells, and tissues.

 [0025] The specimen applied to the reaction apparatus 2 can contain such a reaction component as a test component. Whole blood, serum, urine, feces, saliva, sputum and other biological samples collected from humans and non-human animals; cell follicle cultures; samples of viruses, bacteria, molds, yeasts, plants, animals, etc .; microorganisms Samples that may contain or contain other samples that may contain nucleic acids, proteins, etc.

 [0026] The degree of purification and processing of the test component contained in the sample is not particularly limited. That is, the specimen may be an unpurified sample derived from a living body, or may be appropriately separated and purified, or may contain a test component labeled for detection or the like.

 [0027] In the following description, the reaction apparatus 2 will be described as performing a solid-liquid reaction. The reaction apparatus 2 includes a reagent storage unit 10, a sample introduction unit 20, a concentration unit 40, and a solid-liquid reaction unit that performs a solid-liquid reaction as a final liquid medium reaction in the reaction device 2. 50, a flow path system 60 that communicates each of the above sections so that the reaction can be performed, and a fluid drive section 80 for allowing a liquid or gas to flow through the flow path system 60. In addition, depending on the degree of sample purification and the solution required for the reaction, a purification unit 30 can be provided for sample purification, purification after pretreatment, and the like.

[0028] In the present reactor 2, it is preferable that at least a part of each of the above parts is provided in the solid phase body 100. The solid phase body 100 can be a cube, a rectangular parallelepiped, or an indeterminate three-dimensional shape, but it is a flat body considering the fact that the flow path system 60 is roughly arranged on the same plane. It is preferable. Fig. 2 As shown in the above, it can be a laminate of two or more layers. By doing so, a chamber and a channel system 60 for the reagent storage unit 10, the concentration unit 30, the solid-liquid reaction unit 50, etc. can be easily constructed.

 [0029] Such a solid phase 100 is not particularly limited, and one or more materials can be used in combination. For example, in addition to glass, silicon, ceramics, glass ceramics, plastics such as PMMA, PDMS, metals, and the like can be used. Further, the constituent material of the flow path system 60 can be selected in consideration of the liquidity of the liquid. For example, the constituent material may be selected in consideration of liquid repellency such as hydrophobicity and hydrophilicity.

 [0030] In order to form the flow path system 60 and the recesses or through-holes for each part in the solid phase body 100, for example, a manufacturing technique of a micromachine or MEMS (micro Electro Mechanical System) Can be applied. Various etching techniques, lithography techniques, bonding techniques, film forming techniques, precision micromachining techniques using lasers, ultrasonic techniques, and the like can be used in appropriate combinations. It is also possible to use micro-molding technology such as micro-plastic processing technology, micro-mouth injection molding technology, and micro stereolithography technology. It should be noted that the solid phase body 100 may be appropriately selected according to the molding / processing technique used. Further, when each part and the flow path system 60 are formed by laminating the solid phase body 100, the solid phase body 100 and the other flat plate shape patterned with the flow path system 60 as a through-hole or the like. The solid phase body 100 may be bonded or bonded by an appropriate technique such as thermocompression bonding or ultrasonic bonding. In addition, double-sided adhesive or double-sided adhesive tape can be used for bonding.

 [0031] The reaction apparatus 2 preferably has a desktop size as a whole including the control unit 200, and a portion other than the control unit is a general slide glass (76.2 mm X 25.4 mm ) Is preferable. In addition, for example, the solid-liquid reaction part 5 0 preferably has a volume of 100 I or less, more preferably 60 I or less.

 [0032] (Reagent reservoir)

The reaction apparatus 2 of the present invention can include a reagent reservoir 10. Reagent storage The unit 10 can store various reagents in advance, and can be prepared so that the reagents can be supplied to the solid-liquid reaction or the pretreatment thereof. An appropriate number of reagent reservoirs 10 can be provided according to the type of reagent used. The reagent reservoir 10 can include a chamber 12 having an appropriate capacity and an opening 14 communicating from the chamber to the flow path system 60.

 [0033] The reagent storage unit 10 may be pre-filled with a predetermined amount of, for example, a reagent amount used for one reaction, and may be in a cartridge form that can be exchanged for each reaction. Preferably, the reaction apparatus 2 includes the reagent storage unit 10 as a reagent cartridge 16 that can be replaced with a plurality of reagent storage units 10 that are pre-filled with a plurality of reagents used in one reaction. . According to such a cartridge form, a necessary amount of reagent can be prepared easily and reliably. Such a reagent cartridge 16 is preferably mounted so as to be replaceable so as to form a part of the solid phase body 100 constituting the reaction apparatus 2.

 [0034] The reagent reservoir 10 may have a reagent inlet that can be opened and closed, and may have a form that can be repeatedly used so that a reagent can be injected. The capacity of the reagent reservoir 10 is not particularly limited, but the reaction cost can be reduced and the reaction apparatus 2 can be made compact by setting it to a predetermined capacity capable of storing a necessary amount of reagent. You can.

 [0035] The reagent stored in the reagent storage unit 10 varies depending on the solid-liquid reaction to be performed by the reaction apparatus 2, but includes, for example, an enzyme, its substrate, a nucleic acid probe, a diffusion primer, an antigen, an antibody, a labeling reagent, Various reagents (coloring reagent, reaction reagent, etc.) can be mentioned. In addition, as described later, the reagent also includes an adsorption solution, a washing solution, and an elution solution supplied to the monolith column 32 of the purification unit 30.

 [0036] (Sample introduction part)

The sample introduction unit 20 is a part for introducing the sample into the reaction device 2, and when there is a sample amount to be loaded on the reaction device 2 and a reagent supplied together with the sample, the total amount with the reagent is included. It can be provided with a corresponding capacity. The form of the sample introduction unit 20 is not particularly limited, but the chamber 22 having an appropriate capacity and the chamber Can be provided with an opening 24 communicating with the flow path system 60 from. Similarly to the reagent storage unit 10, the sample introduction unit 20 may have a cartridge form that can be exchanged for each reaction, or may have a form that can be used repeatedly by having a sample introduction port that can be opened and closed. Further, it may have a cartridge form having a sample introduction port that can be opened and closed and can be used repeatedly. In the reaction apparatus 2 shown in FIG. 1, the sample introduction unit 20 is a sample cartridge 26 having a sample introduction port. Note that a plurality of sample introduction units 20 may be provided.

[0037] (Purification Department)

 The reaction apparatus 2 can include one or two or more purification units 30. The purification unit 30 can be provided for the separation or purification of the test component in the sample, the separation or purification of the test component of the pretreatment liquid for the reaction, and the like. The purification unit 30 can be a simple filter or column, but preferably a monolith column 3 2 is used. The monolith column 32 is formed of a structure having macropores that are connected to and communicate with each other. The monolith column 32 has a low pressure loss and good liquid permeability, and is therefore preferable for a biological sample that has many contaminating components and tends to have a high viscosity.

 [0038] As the monolith column 32, an inorganic material structure or an inorganic-organic hybrid structure having the above macropores can be used. The monolith column 3 2 can further have micropores inside the macropores. The inorganic material constituting the monolith column 32 is not particularly limited, and silica or glass material can be used. The surface of the macropores of the monolith column 32 can be appropriately surface-treated with a surface-treating ion-exchangeable resin such as octadecyl component.

[0039] The monolithic column 32 can take a form that can be used as a column itself, and the form is not particularly limited. For example, as a disk-shaped body, it can be used alone or in a plurality of layers. Such a monolithic column 3 2 may be formed by bonding a large number of inorganic particles such as ceramics into a predetermined shaped inorganic structure, or may be porous after being fired using a sol-gel method. An inorganic structure such as refined ceramics may be used. The manufacturing method of such a monolithic column 32 is disclosed in Japanese Patent Application Laid-Open No. 2 0 0 2 _ 3 5 0 4 1 2, Japanese Patent Application Laid-Open No. 2 0 3 _ 7 5 4 2 0, Japanese Patent Application Laid-Open No. 2 4 1 6 7, Japanese Patent Application Laid-Open No. 2 0 0 6-2 2 3 9 60, Japanese Patent Application Laid-Open No. 2 0 0 6-3 4 0 6 4 9 and the like.

 [0040] When such a monolith column 32 is incorporated as the purification unit 30 of the reaction apparatus 2, the through-hole direction of the continuous macropores of the monolith column 32 and the flow path should be in the same direction. preferable. In addition, from the viewpoint of restricting the flow direction of the liquid in the monolith column 32, the inner surface of the O-ring is such that the outflow of liquid from the side surface of the monolith column can be suppressed by fitting the monolith column 32 into the O-ring. It is preferable to adhere. The monolith column 32 thus fitted in the O-ring is accommodated in a part of the cavity of the flow path system 60 of the solid phase body 100. At the time of accommodation, it is preferable that the space between the outer peripheral surface of the O-ring and the inner wall of the cavity is sealed by the O-ring itself or other members.

 [0041] The materials of the O-ring include diaryl rubber, hydrogenated diaryl rubber, fluorine rubber, silicone rubber, ethylene propylene rubber, chloroprene rubber, acrylic rubber, butyl rubber, urethane rubber, chlorosulfonated polyethylene, epoxy Chlorhydrin rubber, natural rubber, fluororesin, etc. can be used

[0042] Alternatively, the monolith column 32 may be clamped with a heat-shrinkable tube and housed in the chamber of the solid phase 100. At this time, it is preferable to seal the inner wall of the chamber of the purification unit 30 and the outer peripheral surface of the heat shrinkable tube. As the heat shrinkable tube, acrylic resin, polyelephine resin, fluororesin, silicone rubber, polyvinyl chloride and the like can be used.

[0043] When the purification unit 30 is equipped with the monolith column 32, the test component in the solution to be prepared with the sample or the reaction apparatus 2 is adsorbed to the test component and then adsorbed to the monolith column 32, and then the test is performed using the washing solution. The impurities other than the components can be discharged, and then the test component can be eluted from the monolith column 32 using the eluate. The monolith force ram 3 2 is the reaction in reactor 2 in the following points (1) to (6) It is possible to greatly contribute to the speeding up of the apparatus and the compactness of the apparatus.

 [0044] (1) Since the pressure loss is low and the liquid permeability is good, it is possible to easily pass a specimen or the like that tends to increase in viscosity inherently or due to concentration. Contributes to compacting of equipment in that it is not necessary to lower the viscosity of the solution

(2) Since the macropores are connected to each other and communicate with each other, the liquid is sent to the monolith column 3 2 or discharged from the monolith column 3 2 by pressurizing or sucking the monolith column 3 2. be able to. For this reason, such pressurization or suction facilitates liquid movement in the column 3 2, so that various reagents and solutions can be quickly sent to and discharged from the column 3 2. it can

 (3) It contributes to automation in that the liquid can be supplied and removed by pressurization and suction, and solid-liquid separation is easy in the monolith column 32.

 (4) The test component can be concentrated or dried in the monolith column 32 by sucking a liquid such as a washing solution while the test component is held in the monolith column 32. For this reason, the concentration of the test component can be increased in the eluate.

 (5) By eluting the test component retained in the monolith column 32 with an eluate that does not contain any salt or that has been suppressed to a level that does not interfere with subsequent concentration or drying. The eluate can be highly concentrated or dried. Therefore, the volume of the solution can be effectively suppressed through the reactor 2, and a high concentration can be easily achieved.

 (6) By using the elution solution as a reagent or medium used in the next step, the eluate can be supplied to the next step as it is, and the process can be simplified and speeded up.

[0045] The use of the monolith column 3 2 for the reaction apparatus 2 is effective in suppressing the volume of the column eluate. Therefore, by using the monolith column 3 2 in the reactor 2, the test component can be easily purified while suppressing the amount of the eluate, and the concentration of the test component in the eluate can be adjusted using another column or purification means. Higher than that Can be tempered. In the reaction apparatus 2, the eluate containing the test component at a high concentration is controlled in such a manner, and the pretreatment for the extraction of the test component from the specimen and the solid-liquid reaction is performed by further concentrating or drying the eluate. Thus, all or part of the amount of the eluate that increases can be easily canceled. As a result, it is possible to suppress the amount of liquid used in the subsequent process, and as a result, it is possible to easily maintain a high concentration and a small liquid volume, and to achieve a rapid reaction and a compact apparatus. .

 [0046] From the above, after one or more pretreatment reactions performed using the reaction apparatus 2, purification using the monolith column 3 2 (preparation of the eluate) and concentration or drying of the eluate are performed. By providing a purification unit 30 and a concentrating unit 40, which will be described later, the final volume of the solution containing the test component obtained by various pretreatment reactions can be extremely effectively suppressed. In addition, the amount of liquid used can be suppressed and the subsequent reaction can be accelerated. In addition, each part that performs various processes can be made compact. In addition, the volume of the solution finally supplied to the solid-liquid reaction is reduced and the concentration of the test component is increased, shortening and speeding up the solid-liquid reaction time, and at the same time making the entire device compact. Can contribute greatly.

[0047] In particular, when the test component is a nucleic acid, the monolith column 32 is inorganic to the extent that no inorganic salts are contained or there is no inconvenience in subsequent concentration or drying by, for example, the following method. An eluate in which the salt concentration is suppressed can be obtained. One is to use a chaotropic salt solution such as guanidine hydrochloride or guanidine thiocyanate as the adsorbent for nucleic acid to adsorb the nucleic acid to the monolith column 32, and then an aqueous solution containing alcohol such as ethanol. Wash the monolith column 3 2 with, and elute it with an eluent (preferably water) in which the concentration of inorganic salts such as water is suppressed. The adsorbing liquid and the cleaning liquid can appropriately contain a strong rhodium salt such as potassium acetate (Japanese Patent Laid-Open No. 2005-0 2 2 4 1 6 7). The other is a solution of an antichaotropic salt such as sodium sulfate or ammonium sulfate as an adsorbent, and an aqueous solution containing an alcohol such as ethanol as a washing solution, and a Tris-HCl buffer, phosphate buffer and water. Inorganic salt concentration is suppressed The obtained solution is used as an eluent (preferably water) (Japanese Patent Laid-Open No. 2000-063 4 0 6 9).

 [0048] When the nucleic acid is purified by the monolith column 32, preferably, an aqueous solution of an antichaotropic salt such as ammonium sulfate is used as an adsorbent, an 80% ethanol aqueous solution is used as a washing solution, and water such as distilled water is used as an eluent. To do. By using these various solutions, salts can be effectively excluded from the nucleic acid eluate from the monolith column 32. It is preferable to supply the adsorbed liquid and the cleaning liquid to the monolith column 32 and then remove the liquid from the monolith column 32 by sufficiently sucking. On the other hand, after the eluate is supplied to the monolith column 32, it is preferable that the eluate is retained in the monolith force ram 32 for a while and then the eluate is discharged. By doing so, the recovery rate of the test component can be improved and the concentration of the eluate can be increased.

[0049] In the purification using the monolith column 32, it is preferable to suppress the pressure loss in the monolith column 32. When the pressure loss increases, it becomes difficult to move the liquid not only in the purification unit 30 but also in the entire reaction apparatus 2, and it becomes difficult to perform a solid-liquid reaction substantially. In order to effectively reduce the pressure loss in the monolith column, it is advisable to defoam the column in advance using a vacuum pump or the like before incorporating it into the reactor. The pressure loss in the monolith column is preferably 5 kgf Z cm ² or less. This is because if the pressure loss exceeds this value, the liquid leaks more frequently and is difficult to use. More preferably, it is 2 kgf Z cm ² or less.

 [0050] The purification unit 30 can be applied to purify and separate a test component from a sample, and a pretreatment reaction product after a pretreatment reaction that processes the test component into a state suitable for the reaction. Can be used to purify. In the reaction apparatus 2, such a purification unit 30 can be provided after the sample introduction unit 20. Further, it can be provided in the preceding stage of the concentrating unit 40 described later.

 [0051] (Concentration part)

The reactor 2 can prepare or store a solution prepared for the reaction or its pretreatment, and can concentrate or dry the solution by distilling off the solvent of the solution 1 Alternatively, two or more concentration units 40 can be provided. As shown in FIG. 2, the concentrating unit 40 includes a chamber 42 having a necessary capacity for reaction, an inlet for receiving reagents and solutions from the previous stage, and a discharge port for discharging semi-solutions. 4 4 b. The size of the chamber 42 is not particularly limited, and can be set to a capacity required for pretreatment reaction or the like.

[0052] The concentrating unit 40 is formed so that the solvent of the solution in the chamber 42 can be distilled off to concentrate or dry the solution. Whether the solution is concentrated or dried may be appropriately set as necessary. It may be determined according to the type and concentration of inorganic salts that may be contained in the solution, or may be determined in relation to the reagent to be used later.

 [0053] According to the concentrating unit 40, even if the amount of the solution stored in the concentrating unit 40 or the amount of the reaction solution in the concentrating unit 40 increases, it can be concentrated or dried on the spot. Therefore, all or a part of the solution in the concentration unit 40 can be canceled to suppress the liquid volume and maintain the test component at a high concentration. Further, as described above, it is preferable that the concentrating unit 40 is provided in the subsequent stage immediately after the purifying unit 30 so that the eluate from the purifying unit 30 can be directly flowed in. In this way, even if the liquid volume is increased by purification in the purification unit 30, the entire amount or a part of the obtained eluate can be easily cancelled.

[0054] Therefore, by providing the concentration unit 40, the reaction apparatus 2 can carry out various pretreatment reactions and intended reactions at high concentrations for the test component. According to the present inventors, in a genomic DNA fragmentation reaction carried out as a pretreatment reaction step, at a normal enzyme concentration (1 UZI), the reaction time is about 120 minutes. It is known that the enzyme concentration can be shortened to 1 minute by increasing the enzyme concentration by 10 times. Further, according to the present inventors, in the fragmented DNA labeling step, by increasing the enzyme concentration by 10 times, the labeling time, which was conventionally about 120 minutes, was reduced to about 20 minutes. I know you can. Thus, increasing the concentration of the reagent and the test component can contribute to speeding up the reaction using various liquids as a medium other than the target reaction. [0055] The concentrating section 40 can be formed so that the solvent is evaporated by heating and the appropriately evaporated solvent is exhausted from the chamber 42. The means for heating the inside of the chamber 42 is not particularly limited, and a known heating means such as a Peltier element can be employed. Further, gas such as air may be supplied to the chamber 42 and exhausted, or drying may be performed by sucking the gas in the chamber 42, and heating by heating means may be added thereto. Such supply and suction of gas into the chamber 42 may be performed using a fluid drive unit 80 which will be described separately.

 [0056] As shown in FIG. 2, the concentrating unit 40 is provided with a breathable material 46 that is exposed in the chamber 42 and has a liquid repellency with respect to the solution and can discharge the gas in the chamber 42. It is possible to provide a ventilation part 48 capable of sucking the gas in the chamber 42 through the breathable material 46. By sucking the gas in the chamber 42 from the ventilation part 48, the solvent in the chamber 42 can be distilled off. This makes it possible to concentrate or dry the solution quickly. Even in this case, it is preferable to heat the chamber 42 as appropriate. Further, the position of the solution in the chamber 42 can be easily controlled by sucking the chamber 42 from the vent 48.

 The breathable material 46 is preferably disposed on the lower surface in the direction of gravity where the solution supplied to the concentrating unit 40 comes into contact. Since the air-permeable material 46 is disposed at such a site, it is easy to stir and mix the solution through the air-permeable portion 48, and the air-permeable material 48 can be As a result, the position control of the solution in the concentrating part 40 becomes possible.

 [0058] Further, the air-permeable material 46 may be provided on a part of the lower surface in the gravitational direction in contact with the solution, or may be provided on the whole. When the entire breathable material 4 6 is provided, the position of the solution can be controlled at any part of the entire bottom surface in the direction of gravity. In addition, when a part of the solution is provided, the solution can be reliably fixed at the site.

[0059] The ventilation part 48 is communicated from the breathable material 46 to the fluid drive system 80, for example. The inside of the chamber 4 2 may be formed so as to be suckable from the breathable material 46. Therefore, it is only necessary to communicate with the inside of the chamber 42 through part or all of the breathable material 46. When it is possible to suck the inside of the chamber 4 2 from a part of the breathable material 4 6, the ventilation part 4 may be communicated only in the part or the suction part can be moved. 8 may be formed so as to be movable along the breathable material 46.

 [0060] When the air-permeable material 4 6 is provided on the entire lower surface of the chamber 4 2 in the direction of gravity, and the ventilation portion 4 8 can suck the gas in the chamber 4 2 from the entire surface of the air-permeable material 4 6 In this case, the solvent in the solution in the chamber 42 can be effectively evaporated, and rapid concentration or drying is possible. In addition, even when bubbles are generated in the chamber 42, they can be easily removed.

 [0061] When the gas permeable material 4 6 can circulate the gas bidirectionally, the gas vent 4 8 can supply gas into the chamber 4 2 through the gas permeable material 4 6. . According to such a gas supply mode, since the gas is not supplied through the limited flow path system 60 such as the inlet 44a and the outlet 44b, the solvent can be effectively distilled off.

 As shown in FIG. 2, the concentrating unit 40 may be capable of concentrating or drying the solution in a predetermined part in the chamber 42. If the solution is concentrated or dried at a part set in advance, the reagent used in the subsequent stage can be concentrated at a site where it is supplied from the reagent reservoir 10 or at a site where it can be easily reached. The test component can be reliably dissolved with a small amount of reagent. In addition, the concentration or dryness of the solution may be the outlet 44 b to the post-reaction side of the concentration part, that is, the vicinity of the communication part. In this way, when the solution is moved to the subsequent stage immediately after dissolution in the reagent, even a small amount of the solution can be moved to the subsequent stage without loss.

[0063] The form of the chamber 42 of the concentration unit 40 is not particularly limited. For example, as shown in Fig. 2, the flow direction of the solution in the concentration unit 40 is the longitudinal direction, and the flow direction of the solution The shape should be long and narrow, with the direction perpendicular to You can. By adopting such a configuration, the flow direction of the solution is the longitudinal direction. For example, with the inlet 4 4 a of the chamber 4 2 opened and the outlet 4 4 b closed, the entire bottom surface in the direction of gravity is As the solvent is distilled off, the solution concentrated as a result of the suction of the inside of the chamber 4 2 from the inside of the chamber 4 2 or the suction of the breathable material 4 6 at the end on the rear side in the longitudinal direction into the chamber 4 2 It is concentrated at the rear end of chamber 42. At this time, if the chamber 4 2 has an elongated shape, the concentrated liquid concentrated at the end on the rear side is visible from the outside to the inlet 4 4 a side with the outlet 4 4 b as the base point. Visible as an extending bar. That is, according to such a configuration, the concentrated solution itself can function as an effective indicator of the degree of concentration. As a result, the amount of the concentrated liquid can be easily detected visually or by a detection means such as a CCD, and when the target concentration level or dryness is detected, the concentration operation can be stopped or controlled. . The concentrating section 40 is preferably oval or rectangular.

 [0064] It should be noted that when the concentration unit 40 functions as a concentration indicator, in order to improve the visibility as an indicator, the predetermined elongated oblong shape or rectangular shape as described above may be used. In this case, it may not be appropriate to perform the pretreatment reaction or the like in the concentrating part 40 because of the capacity. In such a case, the concentrating unit 40 functions only for concentrating, and supplies the reagent used in the next step to the condensing unit 40 concentrated or dried to the subsequent reaction unit. The liquid can be sent to 70.

 [0065] The width of the elongate chamber 42 in the short direction is preferably not more than four times the width of the inlet 4 4a and the outlet 4 4b. If it is 4 times or less, it is effective for the solution near the short wall surface with respect to stirring or mixing by sucking the inside of the chamber through the air-permeable material or the valve opening / closing from the vicinity of the inlet and outlet. Can be implemented. More preferably, it is 2 times or less, and more preferably 1 time or less.

[0066] The breathable material 46 and the vent 48 in the concentrating part 40 can also be used as a mixing part for stirring or mixing the solution in the chamber 42. That is, When the concentrating unit 40 is equipped with such a chamber 4 2, the inside of the chamber 4 2 is alternately sucked from the vicinity of the inlet 4 4 a and the outlet 4 4 b through the air-permeable material 4 6. Alternatively, the solution in the chamber 42 can be stirred or mixed by intermittently forming a differential pressure. As a result, various reactions in the concentration unit 40 can be carried out effectively. This is particularly effective when reacting high concentrations of test components with small amounts of reagents. Further, such differential pressure formation in the chamber 42 can promote concentration or drying in the concentration unit 40. In this case, an appropriate number of ventilation portions 48 are provided in a suction portion convenient for stirring the solution in the chamber 42 by forming a differential pressure, and the breathable material 46 in the portion is changed from the chamber 42 to the chamber 42. The inside can be sucked, or one or two or more ventilation portions 48 can be movably provided at an appropriate suction portion.

 [0067] It should be noted that such differential pressure formation is not necessarily limited to that through the air-permeable material 46 and the air-permeable portion 48. It can also be formed by opening / closing a valve provided to the concentrating unit 40 and the operation of the fluid driving unit 80.

 [0068] In this way, the concentration unit 40 can easily concentrate or dry the solution in the reaction line. Therefore, each pretreatment reaction and solid-liquid reaction can be accelerated, and the apparatus can be made compact. Furthermore, various reactions can be performed in the concentration unit 40, and at the same time, the solution can be concentrated or dried, that is, the two steps can be performed in one element. Speeding up of the process and downsizing of the apparatus can be realized. In addition, the concentration unit 40 can perform concentration or drying of the stored solution, reaction and mixing in the reactor 2 when the solution can be stirred and mixed. And mixing, concentration and mixing, and concentration and reaction and mixing). Therefore, by providing the concentration unit 40 capable of stirring and mixing the solution, the reaction can be speeded up more easily and the reactor 2 can be further compacted.

[0069] The concentration unit 40 does not necessarily have to be provided in the reaction apparatus 2 for the purpose of concentration, and is used only for stirring or mixing the solution in the chamber 42. The reaction apparatus 2 may be provided only as a mixing unit.

 [0070] From the above, it is preferable that the concentration unit 40 has the following configuration. In other words, the concentrating unit 40 is exposed in the chamber 42, and is provided with a breathable material 46 that has liquid repellency with respect to the solution and can discharge the gas in the chamber 42, and the gravity in the chamber 4. It has a ventilation part 48 that can be sucked through the breathable material 46 and has the solution concentrated on a predetermined part of the breathable material 46. It can be dried, and the predetermined part can be a reagent storage part from the reagent storage unit 10. Further, the concentrating part 40 has, for example, an elongated shape in which the solution flow direction in the concentrating part 40 is a longitudinal direction and a direction substantially perpendicular to the solution flowing direction is a short direction, and the width in the short direction is The width of inlet 4 4 a and outlet 4 4 b can be 4 times or less.

 [0071] Note that the air-permeable material 46 has liquid repellency with respect to the solution in contact with the chamber 42, so that the liquid tightness in the air-permeable portion 48 can be improved. The air-permeable material 46 has hydrophobicity on the surface layer side exposed in the chamber 42 when the solution is aqueous and has high polarity, and it has low polarity when the liquid is an organic solvent. Such a surface layer side can be provided with hydrophilicity. Therefore, when the solution is aqueous, it is sufficient if the surface layer side has appropriate liquid repellency, such as a surface layer of a hydrophilic material coated with a hydrophobic material. The breathable material 46 may be any gas as long as the gas in the chamber 42 can be discharged. Preferably, the gas can flow in both directions. Furthermore, it is only necessary to have a good exhaust capacity or distribution capacity against an inert gas such as air or nitrogen. Preferably, it has a good exhaust ability or circulation ability for air.

[0072] Examples of such a breathable material 46 include a material in which fine holes (through holes) are formed in an organic material such as plastic or an inorganic material. Such holes can be formed by laser processing or the like. The size of the hole is not particularly limited, but can be appropriately selected within the range of about 1 m to several hundreds / m. Further, as such a breathable material 46, a porous organic material or a porous inorganic material can be used. The average pore size of such a porous membrane is The average pore diameter is about 0.1 m or more and 0.3 m or less when it is desired to control the gas permeability in the film thickness direction. It is preferable that

 [0073] Typical hydrophobic organic materials used for the air permeable material 46 include polytetrafluoroethylene (PTFE), silicone, polyethylene, polypropylene, polyethylene, polystyrene, polyvinyl chloride, polycarbonate, polysulfone, polyethersulfone, Examples thereof include polyarylene terephthalate, polymethylpentene, and 1,3-cyclohexanegene polymer.

 [0074] By selecting the material of the air-permeable material 4 6 constituting the ventilation part 48, the size and number of the holes, the average pore diameter, the porosity, etc. as necessary, the ventilation through the ventilation part 48. It is possible to perform preferable liquid feeding control by adjusting the direction and the air flow rate.

 Note that the air-permeable material 46 can be applied so as to block the opening of the chamber 42 having a concave shape or the like formed in the concentrating part 40. In closing such an opening with the breathable material 46, conventionally known techniques such as adhesion, adhesion, welding, and thermocompression bonding can be used depending on the type of the breathable material 46. .

 [0076] (Other reaction parts)

 The reaction apparatus 2 can include another reaction unit 70 that prepares or stores a solution prepared for a target liquid medium reaction or a pretreatment thereof. That is, the other reaction unit 70 is a solution storage unit that prepares such a solution but cannot concentrate or dry the solution by distilling off the solvent of the solution. Separately from the solid-liquid reaction part 50 due to the relationship between the previous and subsequent processes and the capacity, etc., or concentrating or drying to solidify the reaction part for only the pretreatment reaction. If desired, these other reaction units 70 can be provided. The other reaction part 70 is preferably elliptical or rectangular like the concentrating part 40.

 [0077] (Solid-liquid reaction part)

The reaction apparatus 2 can include a solid-liquid reaction unit 50. As shown in FIG. 3, the solid-liquid reaction unit 50 includes a chamber 52 for reaction, and a solid phase 56 including a cap 5 7 with which a test component in a solution for solid-liquid reaction interacts. it can. This solid phase may be separate from or the same as the solid phase body 100 which can include each part of the reaction apparatus 2. The cartridge is preferably exchangeable for each solid-liquid reaction. Further, the solid-liquid reaction unit 50 can be provided with an appropriate temperature control means. In FIG. 1, the solid-liquid reaction section 50 has a form of a replaceable reaction cartridge 58.

 The caps 57 used in the solid-liquid reaction part 50 are not particularly limited and can be selected according to the type of solid-liquid reaction. If accompanied by nucleic acid hybridization, it should be a probe, clone (array CGH), etc., if PCR, a primer, if antigen-antibody reaction, antigen or antibody, etc. Can do. In the case of nucleic acid hybridization, it can be carried out by a circulation reaction in addition to a stationary reaction.

 [0079] The chamber 52 can include an inlet 5 4a into which a reaction liquid for a solid-liquid reaction is injected and an outlet 5 4b. The shape of the chamber 52 can be appropriately selected according to the type of the solid-liquid reaction and according to the embodiment. Preferably, the solution in the solid-liquid reaction unit 50 is the same as the concentration unit 40. It is possible to form a long and narrow shape in which the flow direction is the long direction and the direction substantially perpendicular to the flow direction of the solution is the short direction. By adopting such an elongated shape, sufficient nucleic acid can be applied to the solid phase surface on which the reaction liquid and capillaries for solid-liquid reaction are immobilized. The solid-liquid reaction part 50 is preferably oval or rectangular. Preferably, the width of the chamber 52 in the short direction is not more than four times the opening width of the inlet 54a and outlet 54b. With such a width, the reaction liquid for solid-liquid reaction can be uniformly injected and discharged, including the vicinity of the short side wall of the chamber, onto the solid surface where the capillaries are fixed, and bubbles remain on the solid surface. Can be avoided. In addition, when the reaction liquid for the solid-liquid reaction is stirred, it can be effectively carried out including the solution near the short wall of the chamber. Preferably, it is 2 times or less, more preferably 1 time or less.

[0080] The solid-liquid reaction unit 50 can take, for example, a cartridge form as shown in FIG. This reaction cartridge 5 8 has Capture 5 7 immobilized. The cover member 5 5 is provided with an inlet 5 4 a and an outlet 5 4 b for forming a chamber 5 2 of a predetermined capacity on the surface of the solid phase 5 6 with adhesive or adhesive It can be mounted and fixed. By adopting such a cartridge form, a solid phase 56 and a cap 57 corresponding to a solid-liquid reaction can be easily prepared.

 [0081] The solid-liquid reaction unit 50 has a solution for solid-liquid reaction in which the amount of liquid is suppressed through the pretreatment reaction step in the purification unit 30 and the concentration unit 40, and the test component is maintained at a high concentration. Supplied. For this reason, the solid-liquid reaction can be carried out in a short time. According to the present inventors, it has been found that the reaction time can be significantly shortened by increasing the concentration of the test component even in the case of the stationary method. For example, in the nucleic acid hybridization by the stationary method, the reaction time may be about 10 times or less by setting the concentration of the nucleic acid as the test component to about 15 to 40 times the normal concentration. I know what I can do. From the above, in the nucleic acid hybridization, the concentration of the nucleic acid as the test component is preferably 0.3 β g Zu I or more. This concentration is 15 times or more of the hybridization concentration in ordinary array CGH, and the nucleic acid hybridization reaction time can be reduced to about 10 times or less.

 [0082] As described above, in the solid-liquid reaction part 50, increasing the concentration of the test component is effective for speeding up the solid-liquid reaction, and also contributes to the compacting of the solid-liquid reaction part 50. . For example, if the nucleic acid concentration is set to about 15 to 40 times as described above, the solution of the chamber 52 in the solid-liquid reaction section 50 is reduced from 40 to 15 minutes. This is because it can be reduced to about one.

 [0083] The reaction apparatus 2 is preferably capable of washing the solid-liquid reaction section 50. In this way, it is possible to increase the accuracy of the experiment by suppressing the human error by automating the sample detection process immediately after the completion of the solid-liquid reaction, and to shorten the analysis time as a whole. The cleaning liquid can be stored in the reagent storage unit 10 in advance.

[0084] The cleaning method is not particularly limited, but the fluid pressure of the cleaning liquid is 30 kPa or more. And are preferred. This is because if the pressure is higher than this pressure, the cleaning liquid flows uniformly on the solid phase surface, including the vicinity of the short side wall of the chamber. Since non-specifically bound nucleic acids can be removed stably, highly accurate results can be obtained.

 [0085] In addition, the solid-liquid reaction unit 50 has the above-described elongated shape because the cleaning liquid can be distributed throughout the chamber 52 even when the cleaning liquid is continuously passed. And cleaning can be performed in a short time. Also, the longer the shape, the faster the flow rate in the chamber when the same amount of cleaning liquid is supplied. The major factor that affects the cleaning result is the fluid pressure, but it also depends somewhat on the flow rate. For this reason, it is possible to perform the cleaning with a small amount of cleaning liquid in a short time under the condition where the liquid passing speed is high.

 [0086] Further, it is preferable that the solid-liquid reaction part 50 can dry the inside of the chamber 52. If the array of capillaries 57 on the solid phase 56 after washing is dried, it can be immediately analyzed using a detection device such as an appropriate array scanner.

 [0087] (Flow path system)

 The flow path system 60 is a piping system that communicates each of these parts and that can circulate liquids such as gas or reagent for drying, concentration, and various solutions by a fluid drive system described later. The flow path system 60 is preferably provided in a solid phase body 100 including the above-described parts. As described above, the flow path system 60 is appropriately combined with micromachine and MEMS manufacturing technologies, various etching technologies, lithography technologies, bonding technologies, film formation technologies, precision micromachining technologies using lasers, ultrasonic technologies, etc. Can be used. It can also be formed using micro-molding technology such as micro-plastic processing technology, micro-injection molding technology, and micro stereolithography technology.

[0088] (Fluid drive unit)

The reaction apparatus 2 of the present invention includes a fluid drive unit 80 for sending and exhausting a specimen, a reagent, a reaction solution in each part, and the like through a flow path system 60. The flow drive unit 80 can simultaneously supply the regas that has sucked the gas in the flow path system 60 and each unit. Furthermore, the fluid drive unit 80 is It also includes various valves provided in association with each part and the flow path system 60.

[0089] The flow of liquid and the flow of gas may be controlled by two or more fluid drive units 80, or may be controlled by using the same fluid drive unit 80. As such a fluid drive system 80, a liquid feed mechanism using a liquid repellent and air permeable material can be adopted in addition to a known microvalve and a diaphragm pump.

[0090] A liquid feeding mechanism using a liquid-repellent breathable material can have the same structure as the air-flowing portion in the concentration portion already described. In the concentration section 40, the suction of the gas in the chamber 42 from the ventilation section 48 is used for the position control of the solution in the chamber 42, but the position control of the solution is controlled by the flow path system 60, etc. Can be carried out as a solution feed. That is, in the flow path system 60 as well, by appropriately sucking the gas in the flow path system 60 from a part of the flow path system 60 through a breathable material having liquid repellency with respect to the solution, By forming a differential pressure in the flow path system 60, the solution can reach the suction point. Therefore, by moving the suction location according to the liquid arrival location in the flow path system 60, the liquid can be sent as a result.

[0091] (Control part)

 The reaction apparatus 2 of the present invention is a control for performing the purification of the sample in each of the above parts, the pretreatment, the flow of the liquid such as the reagent for performing the solid-liquid reaction, the flow of the gas, the opening and closing of the valve, and the temperature control Part 2 0 0 can be provided. Such a control unit 200 may be a part of the reaction apparatus 2, but may be an external computer connected to the reaction apparatus 2. The control unit 20 0 monitors the state of each unit with various sensors, and controls opening and closing of valves on the flow path system 60, gas flow and temperature by the fluid drive unit 80, and the like by a preinstalled program. be able to.

[0092] Next, an example of a process for performing array CGH by extracting DNA from blood using the reaction apparatus 2 will be described with reference to Figs. In the following description, the steps for purifying DNA, fragmenting the purified DNA, labeling, supplying the labeled DNA to a CGH array and performing array CGH are described. Na Each reagent storage section 10 of the reagent cartridge 16 is loaded in the reaction apparatus 2 in a state in which a necessary amount of necessary reagent is filled in advance, and the solid phase 56 in the solid-liquid reaction section 50 is loaded in the solid phase 56. Clone DNA for array CGH is pre-immobilized and prepared. Further, in the following description, the supply of various reagents and the intake / exhaust of gas are performed based on a program stored in advance in the control unit 20 0 and valves in each part of the fluid control unit 80 and the flow path system 60. Control the opening and closing of the.

 [0093] (Sample introduction process)

 The sample introduction process is performed in the sample introduction unit 20. The sample blood and blood are introduced into the sample introduction unit 20 together with the DNA purification solution for adsorption onto the monolith column 32 of the purification unit 30 for DNA purification. The adsorbing liquid may be supplied from the reagent reservoir 10.

 [0094] (DNA purification process)

 Next, a DNA purification step for purifying the DNA in the sample is performed in the purification unit 30 for DNA purification. The liquid mixture of blood and adsorbing liquid is sent to the purification unit 30 so as to be introduced and discharged. D N A, which is the test component in the mixed solution, is adsorbed to the monolith column 32 by the action of the adsorption solution. After that, the cleaning solution for DNA purification passes through the monolith column 32, is discharged, and is sent to the outside of the reaction line. Further, the elution solution for DNA purification passes through the monolith column 32 and is sent so as to be stored in the purification unit 40. The eluate sent to the purification unit 40 contains DNA, which is a test component.

 [0095] (Concentration step 1)

 Concentration step 1 is performed in the first concentration unit 40 in the reaction line in FIG. The solvent of the eluate eluted from the purification unit 30 and stored in the chamber 42 is retained by sucking the inside of the chamber 42 through the ventilation unit 48 through the ventilation unit 48. Leave to dry. In the concentration step 1, it is preferable to heat the inside of the chamber 42.

 [0096] (DNA fragmentation step)

The DNA fragmentation step is performed in the concentrating unit 40 where the concentrating step 1 is performed. fragment In the fragmentation step, the solution for DNA fragmentation is stored in the concentrating unit 40, and the dried product is dissolved to fragment the DNA. DNA fragmentation is carried out by incubation for a predetermined time in the concentration unit 40. The chamber 42 is preferably heated to a temperature suitable for the enzyme reaction. In the concentration unit 40, a DNA fragmentation step can be performed by forming a differential pressure alternately or intermittently in the chamber 40 and mixing them uniformly.

 [0097] (Step of purifying fragmented DNA)

 In the fragmentation DNA purification step, the fragmentation DNA reaction solution mixed with the adsorption solution in the concentration unit 40 is sent to the purification unit 30 immediately after that to carry out. First, the adsorbing liquid is supplied to the concentrating unit 40 and sent to the outside of the line so as to be discarded. After that, the adsorbing liquid is supplied to the refining unit 30 to the mixed liquid of the adsorbing liquid and the fragmentation reaction liquid, and the outside of the line. To liquid for disposal. Subsequently, the cleaning solution is supplied to the refining unit 30 and sent to the outside of the line to be discarded, and the elution solution is supplied to the refining unit 30 and stored in the subsequent concentration unit 40. Deliver liquid. The eluate sent to the purification unit 40 contains fragmented DNA, which is a test component.

 [0098] (Concentration step 2)

 Concentration step 2 is performed in the second concentration unit 40 in the reaction line in FIG. In the concentration step 2, as in the concentration step 1, the inside of the chamber 42 is sucked from the breathable material 46 of the concentration unit 40 through the ventilation unit 48 to be dissolved from the purification unit 30. Remove the solvent from the eluate stored in 1 and 2 and dry. The concentrated portion 40 is preferably heated.

 [0099] (Labeling step)

The labeling step is performed in the concentrating section 40 where the concentrating step 2 has been performed. In the labeling step, liquid is fed so that the labeling solution (reaction buffer) is stored in the concentration unit 40. Then heat the inside of chamber 42 to denature it in the reaction buffer so that the DNA becomes a single strand, and then cool it down quickly. Subsequently, the labeling enzyme solution (enzyme, substrate, and Cy3 label) is fed so as to be stored in the concentration unit 30. After that, the labeling reaction is performed in the chamber 42 at an appropriate temperature. In the concentration unit 40, The labeling step can be performed by forming a differential pressure alternately or intermittently in the chamber 40 and mixing them uniformly.

 [0100] (Purification process of labeled DNA)

 In the purification process of the labeled DNA, the labeling reaction solution is sent together with the adsorption solution to be stored in the reaction unit 70, and then passed through the purification unit 40 immediately after that and sent to the outside of the line. To do. Subsequently, the cleaning solution passes through the purification unit 40 and is sent to the outside of the line, and the elution solution passes through the purification unit 40 and is stored in the concentrating unit 30 immediately thereafter. Deliver liquid. The eluate sent to the purification unit 40 contains labeled DNA, which is a test component.

 [0101] (Concentration process 3)

 Concentration step 3 is performed in the third concentration unit 40 in the reaction line in FIG. Concentration step 3 is eluted from the purification unit 30 immediately before aspirating the inside of the chamber 42 through the ventilation unit 48 from the breathable material 46 of the concentration unit 40 as in the concentration step 1. Evaporate the solvent from the eluate stored in chamber 42 and concentrate to about 1 in 10 minutes. In this concentration step 3, the next step is a solid-liquid reaction process, so it is sufficient to concentrate to an appropriate reaction volume for carrying out the solid-liquid reaction. It may be preferable to do so.

 [0102] (Array C G H reaction solution preparation process)

 Prior to the array C G H reaction, the step of preparing the reaction solution is performed in the concentration section 40. The concentrated labeled DNA solution is fed so that the solution containing the necessary components for array CGH is stored in the concentration unit 40. In the concentration unit 40, a reaction solution can be prepared by forming a differential pressure alternately or intermittently in the chamber 40 and mixing them uniformly.

 [0103] (Array C G H reaction process)

 The array C GH reaction solution prepared in the concentration unit 40 is fed so as to be stored in the solid-liquid reaction unit 50, and then hybridization is performed for a predetermined time under predetermined conditions.

[0104] (Washing process) After performing hybridization for a predetermined time, the reaction solution is discharged from the chamber 52 and discarded outside the line, and the washing solution is supplied so as to pass through the solid-liquid reaction unit 50.

 [0105] (Drying process)

 After discharging the cleaning solution, gas (air) is supplied to the solid-liquid reaction unit 50 to dry the array of capillaries 57 on the solid phase 56.

 [0106] Thereafter, the reaction cartridge 58 is removed from the reaction apparatus 2, and the fluorescence based on the label is observed with an appropriate scanner. The detection means is not particularly limited. In the case where a label is assigned, any detection means capable of detecting the assigned label may be used, and the hybridization product can be detected electrically or chemically without using the label. It may be a thing.

 [0107] As described above, according to the reaction apparatus of the present invention, the subsequent reaction is performed while all or part of the amount of liquid added by various processes in the apparatus is canceled by the concentrating unit. be able to. As a result, even if multiple steps are performed in the apparatus, the increase in the liquid volume can be suppressed and the concentration of the test component can be increased. At the same time, the response of the entire device can be accelerated and compact. In addition, since it can be compactly automated, biochemical information including genetic information can be easily and accurately acquired.

 [0108] Further, by providing a concentration part capable of sucking the gas in the chamber through the air-permeable material and the ventilation part, various solutions can be concentrated or dried and reacted, so that the process can be performed quickly. And making the device more compact, and by allowing the solution to be stirred and mixed in the concentrating part, further speeding up the process and making the device more compact.

[0109] Although the reaction apparatus of the present invention has been described above, based on the above embodiment, in other embodiments of the present invention, only the various pretreatment reaction steps are performed up to the solid-liquid reaction section. A pre-processing device is also included. According to this pre-processing apparatus, complicated processing can be performed easily and compactly. The pre-processing device has the various types described above. The reaction apparatus of the present invention can be carried out in a form in which the solid-liquid reaction part is not provided.

 [0110] In addition, another embodiment of the present invention may be configured to include only each unit that performs the steps after the solid-liquid reaction. For example, a solid-liquid reaction part that performs a solid-liquid reaction process, a solid solution reaction part that performs a cleaning process after the solid-liquid reaction, and a fluid drive part that passes the cleaning liquid to the solid-liquid reaction part. A liquid reactor may be used. Further, the solid-liquid reaction apparatus may be configured to allow a gas for drying the array or the like in the solid-liquid reaction part to flow through the fluid driving part. Furthermore, other embodiment of this invention can also be set as the structure provided only with each part which implements the washing | cleaning process after a solid-liquid reaction, and a drying process.

 [0111] Further, another embodiment of the present invention includes a reaction apparatus not provided with a control unit.

. The control unit may be installed separately from the reaction device.

 [0112] Furthermore, in the above description, most of the concentrating unit 40 also functions as a reaction field in which a new reagent is added to the concentrate or dried product to react, but the concentrating unit 4 It is possible to make it function as 0 only, and to provide a reaction part 70 for the concentrated or dried product separately. In particular, when the concentration unit 40 is used as the concentration indicator at the same time, the reaction in the next stage of the concentrate or the dried product is performed by supplying the reagent of the next step to the concentrate or the dried product and dissolving it after the dissolution. It is preferable that the reaction is performed at the reaction section 70.

 [0113] According to the reaction method of the present invention, all or a part of the amount of liquid added by various pretreatments such as purification, fragmentation, and labeling is subjected to concentration in the concentration step, and the liquid used for starting the subsequent stage. An increase in the amount can be suppressed. For this reason, even when multiple steps are required to reach the final reaction step, the increase in the liquid volume is suppressed, and the concentration of the test component is increased, resulting in the speeding up of the intended final reaction. it can. As a result, biochemical information including genetic information that generally requires multiple steps can be easily and accurately acquired.

[0114] Further, another embodiment of the present invention includes an embodiment as a pretreatment method in which only the pretreatment process leading to the final reaction process is performed. [0115] In the above description, the reaction apparatus and the reaction method of the present invention are used as a reaction apparatus having a solid-liquid reaction part and a solid-liquid reaction step in which a reaction using a liquid as a medium is a solid-liquid reaction. Although described, the reaction apparatus and the reaction method of the present invention are not limited to the solid-liquid reaction apparatus and the solid-liquid reaction method. Depending on the target reaction, the solid-liquid reaction part and the solid-liquid reaction process can be implemented as a reaction part and a reaction process that perform normal liquid medium reactions in addition to interfacial reactions such as gas-liquid and liquid-liquid reaction. can do.

 [0116] The reaction method of the present invention can take the following embodiments.

 (1) A reaction method using a liquid as a medium,

 Using a reactor equipped with a concentration unit capable of concentrating or drying the solution by distilling off the solvent of the solution while preparing or storing the solution prepared for the reaction or its pretreatment,

 A solution preparation step of preparing a solution prepared for the reaction or its pretreatment; a purification step of purifying the solution to obtain a purified solution;

 A concentration step of concentrating or drying the purified liquid in the concentration section;

 Comprising a method.

 (2) The method according to (1), wherein the purified solution is an eluate from a purification column.

(3) The method according to (2), wherein the purification column is a monolith column.

 (4) The method according to any one of (1) to (3), wherein the concentration step is a step of concentrating or drying the purified liquid to a predetermined part in the concentration unit.

 (5) The method according to any one of (1) to (4), wherein the concentration step is a step of drying the purified solution.

 (6) The method according to any one of (1) to (5), wherein the concentration step is a step involving suction or heating in the concentration unit.

 (7) After the concentration step,

In the concentration section, the concentrated or dried solid product The method according to any one of (1) to (6), wherein a subsequent solution preparation step of preparing a solution prepared for the solid-liquid reaction or its pretreatment is performed.

 (8) The method according to any one of (1) to (7), wherein the reaction using the liquid as a medium is a solid-liquid reaction.

 (9) The concentrating unit includes a breathable material that is exposed in the concentrating unit and has liquid repellency with respect to the reaction solution and can discharge the gas in the concentrating unit, and the gas in the concentrating unit is used as the breathable material. And a ventilation part that can be sucked through,

 The method according to any one of (1) to (8), wherein the concentration step is performed by gas suction from the ventilation section.

 (10) The concentration unit can mix the solution by forming a differential pressure in the concentration unit,

 After the concentration step,

 In the concentration section, a solution prepared for the reaction or the pre-treatment is prepared for the concentrate or the dried product by the concentration or drying, and the solution prepared at the latter stage is prepared while forming the differential pressure and mixing. The method according to any one of (1) to (9), wherein the step is performed.

 (1 1) The concentrating unit is a gas-permeable material that is exposed in the concentrating unit and has liquid repellency with respect to the reaction solution and can discharge the gas in the concentrating unit, and the gas in the concentrating unit The method according to (10), further comprising: a vent part that can be sucked through a material, wherein the subsequent solution preparation step is a step of forming the differential pressure by suction from the vent part.

 (1 2) The method according to any one of (1) to (1 1), wherein the purification step is a step of obtaining a purified solution that is an aqueous solution.

 (1 3) The method according to any one of (1) to (12), wherein the test component of the reaction is a nucleic acid.

 (14) The method according to any one of (1) to (13), wherein the reaction is a solid-liquid reaction.

According to the reaction apparatus and reaction method of the present invention, various disease genes, constitutional genes, etc. In addition to the nucleotide sequence, mutation, deletion, amplification, and expression status of any gene, individual patient data regarding SNPS, etc. can be easily obtained. Furthermore, as a result, diagnosis such as drug sensitivity, side effects, disease name, treatment prognosis, etc. can be made based on individual biochemical information, so that personalized medicine can be promoted and preventive medicine can be promoted. .

 [0118] The reaction apparatus and reaction method of the present invention can be used as various genetic test apparatuses and genetic test methods, or pretreatment apparatuses and pretreatment methods thereof. For example, clinical examination, diagnosis, drug screening, medicine, genetic identification in forensic medicine, environmental analysis, food inspection, forensic medicine, brewing, fishery, livestock, agricultural production, genetic testing in fields such as agriculture and forestry It can be used as a chemical inspection apparatus and inspection method or a pretreatment apparatus and pretreatment method thereof.

 Example 1

 [0119] In this example, the reaction apparatus schematically shown in Fig. 1 was manufactured, and extraction, fragmentation, labeling and hybridization of various solutions of hybridization using the array CGH as an application system and reaction were performed. Confirmed shortening of time. In addition, the amount of DNA recovered in each reaction step was evaluated.

 [0120] Fragmentation>

 Fragmentation was performed in a concentration section where fragmentation reaction and concentration of the reaction solution were possible. Fragmentation was performed in the following steps. Purification after DNA fragmentation was carried out in a purification section equipped with a monolith column (ø2mmX 2.6 mm).

 1. Set the sample, reagent, etc. in the specified place of the reactor.

 'Fragmentation reaction sample (0.24 gZ1 0 L) * Status after DNA extraction ■ Fragmentation buffer (0.22 L)

 ■ Fragmentation enzyme (1. 94 L)

 'Adsorption buffer (1 0. 8 L) (4 M ammonium sulfate aqueous solution) ■ Washing buffer (84 L) (80% ethanol aqueous solution)

 ■ Elution buffer (5 L) (water)

2. Dry the reaction sample in the concentration section 3. Transfer the DNA fragmentation solution (fragmentation buffer and fragmentation enzyme) to the concentration section.

: 曰 A

, Higuchi

 4. Enzymatic reaction at 37 ° C for 1 minute

 5. Dispose of the adsorption field buffer together with the enzyme reaction solution through a monolithic column.

 6. Pass the wash buffer through the monolith column and discard

 7. Pass the elution buffer through a monolith column and collect it in the concentration section.

 8. The eluate from the column is dried for the labeling process, which is essentially the next step, but the eluate is collected and the absorbance is measured for the evaluation of the monolithic column.

9. Dry the eluate in the concentration section

 , Δ Z

 The eluate that was fragmented and purified by the reactor was applied to the label in the reactor. The labeling reaction was performed in a concentrating part capable of concentrating the reaction solution. A monolithic column (02mmX 4.6mm) was used for purification after labeling. Labeling was performed in the following steps.

 1. Set the sample, reagent, etc. in the designated place of the pretreatment device.

 ■ Dry product (Fragmented DNA) (0.19 g) * Dry state after fragmentation ■ Labeling buffer (7. 07 L)

 ■ d CTP M i x t u r e (1.9 L)

 ■ Cy 3-d CTP (1.1 4 L)

 'Labeling enzyme solution (1.4 L)

 'Adsorption buffer (57. 5 L) (4 M ammonium sulfate aqueous solution) ■ Washing buffer (300 L) (80% ethanol aqueous solution)

 ■ Elution buffer (18 L) (water)

 ■ 20 X Co t-1 DN A (0.44 L)

 ■ MM # 1 (5.6 L)

 ■ 5 X y e a s t t RN A (0. 1 6 L)

 ■ 20% S DS (1.6 L)

2. Transfer and mix the labeling buffer to the concentration section 3. Thermal denaturation 95 ° C, 5 minutes

 4. Cooling 4 ° C, 10 minutes

 5.d CTP M i xt ure, Cy 3_d CTP, and labeling enzyme are sent to the concentration unit.

 6. Enzymatic reaction at 37 ° C for 40 minutes

 7. Dispose of the adsorption buffer together with the enzyme reaction solution through a monolith column

8. Pass the wash buffer through the monolith column and discard

 9. Pass the elution buffer through a monolith column and collect it in the concentration section.

 1 0. Although the eluate is dried for the next reaction step, the eluate is collected and the absorbance is measured for monolithic column evaluation.

 1 1. Dry the eluate in the concentration section

 1 2. 20xCo t _ 1 DNA, MM # 1, 5 X y e a s t tRNA, 20% S D S is sent to the concentration section to prepare the reaction solution

 The DNA concentration in this hyperprecipitation solution was 15 times that of the comparative example of the conventional method.

[0122] <Solid-liquid reaction (hybridization)>

 A solid-liquid reaction chamber with a volume of 8 I having the configuration shown in FIG. 3 was prepared. As the solid phase, a CGH array in which the target clone DNA was immobilized in advance was used. Next, this solid-liquid reaction chamber was set in a reaction apparatus equipped with a microvalve, a diaphragm type micropump, and a cleaning liquid tank. At this time, each of the inlet and outlet of the solid-liquid reaction chamber was joined so as to communicate with the microphone port valve of the reaction device via an O-ring.

 * Micro valve (LV-2—MF F, manufactured by Takasago Electric)

[0123] After injecting the hybridization solution into the reaction chamber, hybridization was performed under the following conditions.

 Reaction time: 4 hours

 Reaction temperature: 42 ° C

Reaction volume 8 \ [0124] After the reaction time, washing was performed under the following conditions.

 Cleaning time: 7 minutes in total

 Washing temperature: 42 ° C

 Cleaning fluid volume: 500 I each

 Cleaning method: Liquid feeding cleaning (Liquid for each cleaning liquid is injected from the inlet of the solid-liquid reaction chamber and discharged from the outlet)

 Cleaning solution:

 (1) 2 550 solution delivered for 1 minute

 (2) 50 ° C 50% formamide Z2 X SSC solution for 1 minute

 (3) 50 ° C 2 X SSCZ 0. 1% SDS solution for 2 minutes

 (4) PN B u f f e r solution for 1 minute

 (5) 2 550 solution for 1 minute

 [0125] After washing, air was blown from the micropump for 1 minute to dry. Thereafter, fluorescence measurement was performed in a scandal (A g i lent T e c h n o l o g i e s ^ M i c r o r r a r y S c a n n e r). The fluorescence signal intensity was digitized using a numerical analysis software (GenePix.Pro, manufactured by Axon). In addition, a numerical analysis was performed.

[0126] Next, as a comparative example for DNA fragmentation and labeling, DNA was fragmented and labeled according to the procedure described in Cell Engineering Vol. 23 No. 3 2004. In addition, ethanol precipitation was used for purification. Further, the labeled DNA thus obtained was mixed with the following reaction reagent to obtain a hybridization solution (120 I).

 MM # 1: 84 I

 Y e a s t t RN A: 1 2 I

 20% S DS: 24 μI

[0127] For the hybridization, a solid-liquid reaction chamber with a volume of 120 I having the configuration shown in Fig. 3 was separately prepared. The same CGH array as in the example was used. [0128] After injecting the hybridization solution into the reaction chamber, the reaction chamber was set in an oven (FV-320, manufactured by ADVAN TEC), and hybridization was performed under the following conditions.

 Reaction time: 48 hours

 Reaction temperature: 42 ° C

 Reaction volume: 1 20 I

[0129] After the reaction time, washing was performed under the following conditions.

 Cleaning time: 70 minutes in total

 Cleaning temperature: room temperature and 50 ° C

 Cleaning fluid volume: 250 m I each

 Cleaning method: immersion cleaning

 Cleaning solution:

 (1) rinse at room temperature with 2 XSSC solution

 (2) Immersion cleaning for 15 minutes in 50 ° C, 50% formamide Z2 X S S C solution

(3) 50 ° C, 2 XSSCZ 0.1% 30% immersion cleaning in 1% SDS solution

 (4) Immersion cleaning at room temperature for 15 minutes in PN buffer solution

 (5) Wash for 5 minutes in 2 XSSC solution at room temperature

 [0130] After washing, the array was centrifuged (1 000 rpm 3 minutes) and dried, followed by fluorescence measurement and analysis in the same manner as in the Examples.

 [0131] <Recovery result of fragmented DN A>

Table 1 shows the recovery results for the amount of genome (0.24 g) used in the fragmentation process. As shown in Table 1, in this example, 0.19 g (recovery rate of about 81%) could be stably recovered, which was equivalent to the comparative example of the conventional method. From the above, it was found that DNA fragmentation and fragmentation DNA recovery at the same level as the conventional method can be performed. In addition, the time required for the DNA fragmentation step in the conventional method was 135 minutes, but in this example, it was shortened to 9 minutes. Therefore, according to the reactor of the present example, it was found that the intended amount of DNA can be recovered and the process can be shortened remarkably. [table 1]

 [0132] <Recovery result of labeled DNA>

 Table 2 shows the recovery results for the amount of DNA (0.19 g) used in the labeling process. As shown in Table 2, in this example, 2.5 μg (recovery rate about 13 times) could be recovered stably, which was equivalent to the conventional method of Comparative Example. From the above, it was found that according to the present example, it is possible to recover the DNA label and labeled DNA at the same level as the conventional method. In addition, in the conventional method, the labeling process time was 270 minutes, but in this example, it was shortened to 70 minutes. Therefore, according to the reactor of this example, it was found that the intended amount of labeled DNA can be recovered and the process can be significantly shortened.

 [Table 2]

 [0133] <Hybridization result>

FIG. 5 shows a comparison result of the fluorescence signal ratio I og ₂ (C y 3 / C y 5) by the array CG H according to Example 1 and the comparative example. As shown in Fig. 5, the fluorescence signal ratio L og 2 (C y 3ZC y 5) with the conventional method shows a correlation coefficient of 0.857 3, and a data-correlated result was obtained. Fig. 6 shows a scan image according to Example 1. As shown in Fig. 6, high primula, dirt, etc. were not recognized in the scanning image.

[0134] Based on the above, increase the DNA concentration by 15 times during hybridization It was found that the reaction time of the hybridization can be reduced from 48 hours to 4 hours. That is, it has been found that increasing the concentration of the test component is extremely effective for speeding up the solid-liquid reaction such as hybridization. It was also found that increasing the concentration of the test component is effective for compacting each part of the device. In addition, it was found that the cleaning time can be reduced from the conventional 70 minutes to 7 minutes by canceling the diffusion rate limiting of the cleaning by liquid feeding cleaning instead of immersion cleaning o

 Example 2

 [0135] In this example, a test was conducted to shorten the process time by increasing the enzyme concentration in the DNA fragmentation process.

[0136] DNA fragmentation>

 First, the following reagents were placed in a 0.5 ML microtube and mixed (enzyme concentration: 5 UZ I).

 ■ Genomic DNA K562 (Cosmo Bio, product number D 1 255820): 1 μ

S (1 J "; 1 g / de)

 ■ Dpn I I buffer (D pn I I uf er & enzyme is made by N EW ENGLAND Bio Labs I NC.): 2 L

 ■ Dpn 11 enzyme: 1 00 U (1 0 L; 1 0 U / μL)

 ■ Sterile water: 7 fl L

 [0137] Next, this mixture was incubated at 37 ° C, 10 minutes, 5 minutes, 1 minute using a heat block (Block Incubator BI _525, manufactured by ASTEC), and PCR purification Kit ( QI AG EN). Furthermore, this purified product was subjected to electrophoresis (2% agarose, 1 Kb

 DNA Ladder).

 [0138] As a comparative example, the same as the above example except that the amount of sterilized water is 16 μI, the enzyme is 10 U, the incubation time is 2 hours (enzyme concentration 0.5 UZ I). Then, the D Α Α fragmentation step was performed.

[0139] In the examples, the enzyme concentration was 10 times that of the comparative example. 1 minute in the example It was possible to fragment DNA in the same manner as in the comparative example. From the above, it was found that DNA can be fragmented in a very short time by increasing the enzyme concentration.

 Example 3

 [0140] In this example, a test was carried out to shorten the process time by increasing the enzyme concentration in the DNA labeling process.

[0141] Gu signs>

 The following reagents were placed in a 0.5 M L microtube and mixed. 2.5 X R a n d om

 Primers Solution was manufactured by Rogen.

 (B i o P r i m e Array C G H G e n o m i c L a b e l i n g S y s t em; Part No. 1 8095— 0 1 1)

 ■ Fragmentation D NA: 0.5 g (1 2 // L; 1 g / 24 // L)

 • 2.5 X b u f f e r mix: 20 L

 [0142] Next, this mixed solution was incubated at 99 ° C for 5 minutes using a heat block (block incubator B I — 525, manufactured by ASTEC), and ice-cooled for 10 minutes. Furthermore, 10 xdCT P Nuclte o te dM ix 5 L, Cy 3 _dCT P 3 L, sterilized water 9 β I and E xo -K I e n w f r a g m e n t 1 L were added and mixed. 10 xdCT PN ucleotide M ix &. E xo— KI enowfra gm ent la »I nv ι trogen, (B io Prime Array CGHG en om ic Labelin Syst em; Part No. 1 8095— 0 1 1) C y 3— dCT P was manufactured by GE hea Ithcare (part number PA 5303 1).

 [0143] This mixture was incubated in a heat block at 37 ° C for 1 minute, 5 minutes, 10 minutes, 12 minutes, 15 minutes and 20 minutes. The reaction product was purified by PCRpurification kit, and the absorbance (A260; DNA amount, A530; Cy3 labeled amount) was measured with a spectrophotometer.

[0144] As a comparative example, E xo _K lenowfra gm ent 10 L was added. The labeling step was carried out in the same manner as in the above example except that the enzyme amount was 10 times and the incubation time was 2 hours, and the final DNA amount and Cy 3 labeling amount were measured.

 [0145] In the examples, the enzyme concentration was 10 times that of the comparative example. In the example, DNA could be fragmented in an incubator for 20 minutes as in the comparative example. From the above, it was found that DNA can be labeled in a very short time by increasing the enzyme concentration.

 [0146] This application is a US patent application number filed provisionally in the United States on April 18, 2006.

 60/792, 620, US Patent Application No. 60Z792, 982 filed provisionally in the United States on April 1, 2006 and Japanese Patent Application Patent Application 2006—26921 7 filed on September 29, 2006 All the contents of this document are included in this specification as the basis for priority claim.

Claims

The scope of the claims

 [1] A reaction apparatus for reaction using a liquid as a medium,

 A reagent reservoir for supplying a reagent used for the reaction or its pretreatment, a sample introduction unit for supplying a sample,

 A concentration unit capable of concentrating or drying the solution by distilling off the solvent of the solution while preparing or storing the solution prepared for the reaction or its pretreatment, and a reaction unit for performing the reaction When,

 A flow path system that communicates each of the parts to enable the reaction;

 A fluid drive unit for flowing the liquid or gas for the reaction in the flow path system;

 Comprising a reaction device.

 [2] The apparatus according to [1], wherein the solution in the concentration part is dried in the concentration part.

[3] The concentration or drying of the solution involves suction or heating in the concentration section.

 The apparatus according to 1 or 2.

[4] The concentration unit has a flow direction of the solution in the concentration unit as a longitudinal direction, a direction substantially orthogonal to the flow direction of the solution as a short direction, and the short direction is a width of a solution supply site And the apparatus according to any one of claims 1 to 3, wherein the apparatus has a width that is not more than four times the width of the discharge site of the solution.

[5] The solution in the concentration part can be concentrated or dried to a predetermined part of the concentration part.

The apparatus according to any one of claims 1 to 4.

[6] The apparatus according to [5], wherein the dry or concentrated portion of the solution is in the vicinity of a communication portion of the concentrating portion to the reaction post-stage side.

[7] The apparatus according to claim 5 or 6, wherein the dry or concentrated part of the solution is a reagent storage part from the reagent storage part.

[8] The concentration unit has a longitudinal direction as a flow direction of the solution in the concentration unit, a short direction as a direction substantially perpendicular to the flow direction of the solution, and the short direction is a width of a solution supply site The apparatus according to claim 7, wherein the apparatus has a width that is not more than four times the width of the discharge site of the solution. The apparatus according to any one of claims 5 to 8, wherein a concentration part of the solution remaining in the concentration part is visible from outside the concentration part.

 The concentration unit includes

 A breathable material that is exposed in the concentration section and has liquid repellency with respect to the reaction solution and can discharge the gas in the concentration section; and a ventilation section that can suck the gas in the concentration section through the breathable material. The reaction apparatus according to claim 1, comprising:

 The reaction apparatus according to claim 10, wherein the concentrating unit includes the air-permeable material on at least a part of a lower surface in a gravitational direction in contact with the reaction solution supplied to the concentrating unit.

 The apparatus according to claim 11, wherein the concentrating portion includes the air-permeable material on substantially the entire lower surface in the gravitational direction.

 The apparatus according to any one of claims 10 to 12, wherein the concentration unit includes the ventilation unit so that gas in the concentration unit can be sucked from the entire surface of the breathable material exposed in the concentration unit. .

 The said concentration part is equipped with the said ventilation part so that the gas in the said concentration part can be attracted | sucked from a part of said breathable material exposed in the said concentration part, The statement of any one of Claims 10-12. apparatus.

 The concentration unit includes

 A gas permeable material that is exposed in the concentrating part and has a liquid repellency to the reaction liquid and capable of discharging the gas in the concentrating part; A breathable portion that can be sucked through the breathable material,

 The solution in the concentration part can be concentrated or dried to a predetermined part of the breathable material,

 The apparatus according to claim 1 or 2, wherein the predetermined part is a reagent storage part from the reagent storage part.

The concentration part has a longitudinal direction as a flow direction of the solution in the concentration part, The direction substantially perpendicular to the flow direction of the solution is the short direction, and the short direction is a width of 4 times or less the width of the solution supply site and the width of the solution discharge site, The apparatus according to claim 15, wherein a concentration site of the solution is visible from outside the concentration unit.

[17] The solution can be mixed by forming a differential pressure in the concentration section.

 The apparatus in any one of 1-16.

[18] The differential pressure is formed by suction of the gas in the concentrating part through a breathable material that is exposed in the concentrating part and has liquid repellency to the reaction liquid and can discharge the gas in the concentrating part. The apparatus of claim 17.

[19] The apparatus according to any one of claims 1 to 18, further comprising a purification unit that purifies a test component in a solution prepared for the reaction or a pretreatment thereof.

[20] The purification section includes a monolith column.

 The apparatus according to claim 19, wherein the monolith column is capable of eluting the test component using water as an eluent after adsorbing the test component.

[21] The apparatus according to [19] or [20], wherein the eluate containing the test component purified by the purification unit can be concentrated or dried in the concentration unit.

[22] The device according to any one of claims 19 to 21, wherein the test component is a nucleic acid.

[23] The apparatus according to any one of claims 19 to 22, wherein the solution or the reagent can be introduced into the monolith column in a pressurized or reduced pressure state.

[24] The reaction unit has a flow direction of the solution in the reaction unit as a longitudinal direction, a direction substantially orthogonal to the solution flow direction as a short direction, and the short direction is a width of a solution supply site And the apparatus according to any one of claims 1 to 23, wherein the apparatus has a width that is not more than four times the width of the discharge site of the solution.

25. The apparatus according to claim 24, wherein the test component is a nucleic acid, and the reaction in the reaction part is a nucleic acid hybridization.

26. The apparatus according to claim 25, wherein the nucleic acid hybridization is an array CGH.

[27] When the concentration of the nucleic acid as the test component in the reaction part is 0.3 g / l or more, The device according to claim 25 or 26.

 The reaction apparatus according to any one of claims 1 to 27, wherein the reaction using the liquid as a medium is a solid-liquid reaction.