JP2007319097A - Reaction kit - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To prevent the intrusion of a foreign matter into a reaction plate from the outside, and an environmental pollution on the outside. <P>SOLUTION: This reaction kit is characterized by comprising a reaction plate 2 which has a reaction container 4 for causing the reaction of a sample and a reagent container 12 receiving a reagent used for the reaction of the sample and sealed with a film 14 and on whose upper side the members are formed, an injection chip 20 disposed on the upper side of the reaction plate 2, a cover 24 which covers a space on the upper side of the reaction plate 2 and movably supports the injection chip 2 in such a posture that the tip end side and base end side of the injection chip 20 are disposed on the inside and the outside, respectively, and a sample-introducing portion 32b for injecting the sample from the outside into the inner space through a sample-introducing port 33b closably disposed at a portion of the cover 24, wherein the sample-introducing port 33b is formed from an elastic member 33c which can be penetrated with an injector having a sharp tip and can close the penetrated hole, when the injector is pulled out after the penetration. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a reaction kit suitable for performing various analyzes and analyzes in the field of medical analysis and chemistry in the fields of biological analysis, biochemical analysis, or chemical analysis in general.

  A micro multi-chamber apparatus is used as a small reaction apparatus used for biochemical analysis or normal chemical analysis. As such an apparatus, for example, a microwell reaction plate such as a microtiter plate in which a plurality of wells are formed on a flat substrate surface is used.

In the conventional microwell reaction plate, the upper surface of the reaction plate is open to the atmosphere when used. Therefore, foreign matter may enter the sample from the outside, and conversely, the reaction product may contaminate the external environment.
Accordingly, an object of the present invention is to provide a reaction kit that can prevent foreign substances from entering from the outside of the reaction plate and environmental pollution to the outside.

  The reaction kit of the present invention comprises a reaction plate having a reaction vessel for causing a sample to react on the surface side, a dispensing chip disposed above the surface side of the reaction plate, and an upper space on the surface side on the reaction plate A cover that movably supports the dispensing tip so that the tip end is inside the space and the base end is outside, and the sample introduction is provided in a part of the cover so as to be sealed A sample introduction unit for injecting a sample into the space from the outside through the mouth.

  When putting a sample into the sample inlet for injecting the sample into the space from the outside and closing it tightly, you must open the lid of the sample inlet once, but open the lid, dispense the sample and close the lid There is a risk that foreign matter may be mixed in from the outside, and opening and closing the lid is troublesome, so that the sample inlet of the present invention can be penetrated by a sharp dispensing device with a sharp tip, and after the penetration When the dispensing instrument is pulled out, the through hole is constituted by an elastic member that can be closed by elasticity.

  A preferred example of the sample introduction part forms a container, the sample introduction port is on the side of the container, and has an opening at the top of the container, and the sample pretreatment liquid or reagent is enclosed in advance in the container It is what has been.

  The opening may be sealed with a cover film so that the sample does not spill out of the container in order to prevent drying or when the reaction kit is accidentally dropped.

  In order to prevent the sample adhering to the elastic member from flowing out and being contaminated, the sample inlet may be sealed by attaching a seal film to the sample inlet.

  The reagent used for the sample reaction must also be introduced into the space covered by the cover by some method, and the method is not particularly limited. For example, the reagent may be introduced from the sample introduction part together with the sample. Alternatively, it may be introduced in a separate container, or may be accommodated in a reaction plate in advance. In the form in which the reagent is stored in advance in the reaction plate, the reaction plate also includes a reagent container that stores the reagent on the surface side and is sealed with a film. The film covering the reagent container and sealing the reagent can be penetrated by a dispensing tip.

  The space on the surface side on the reaction plate is covered with a cover and shielded from the outside, and the reaction for the sample is performed in that space. Detection of the reaction product after the reaction is also performed in a state where the reaction product is in the cover without taking the reaction product out of the cover. After detection, the reaction kit is discarded while the reaction product remains in the cover. That is, this reaction kit is disposable.

  The dispensing tip may be attached to the tip of the dispensing nozzle. In that case, a nozzle mechanism is separately required for the dispensing operation. Therefore, in order to eliminate the need for such a nozzle mechanism, in a preferred embodiment of the present invention, the dispensing tip is provided with a syringe operated from the outside of the cover, and the dispensing operation is performed by operating the syringe. It can be. When the dispensing tip includes a syringe, since the syringe seals the passage of the dispensing tip, the inside and outside of the space covered with the cover does not pass through the passage of the dispensing tip.

  If the dispensing tip is not equipped with a syringe, it can be sealed with a nozzle mechanism during dispensing operation, but if the dispensing tip is not used, such as during reaction or detection, Note It communicates with the external space through the tip. Even in such a case, as a preferable mode for preventing foreign matter from entering from the outside and preventing the sample and its reaction product from going outside, the dispensing tip has a filter inside the tip portion. Can be.

  When this reaction kit is intended for gene analysis, the reaction plate is preferably provided with a gene amplification section for performing a gene amplification reaction on its surface side. It is preferable that the gene amplification section has a shape suitable for temperature control at a predetermined temperature cycle. The reaction vessel can be formed in such a shape to be a gene amplification section. An amplification vessel may be provided. The gene amplification reaction includes a PCR method and a LAMP method.

The analysis of the reaction product in the reaction vessel can be performed in the reaction vessel or moved from the reaction vessel to another place on the reaction plate.
In a reaction kit in which a reaction product is analyzed in a reaction vessel, the reaction vessel is preferably made of a light-transmitting material so that optical measurement can be performed from the bottom.

  In the reaction kit in which the reaction product is analyzed by moving it from the reaction vessel to another location, the reaction plate further includes an analysis unit for analyzing the reaction product in the reaction vessel on the surface side. Yes.

An example of such an analysis unit is an electrophoresis unit that performs electrophoretic separation of reaction products.
Another example of such an analysis unit is a region where a probe that reacts with a gene when the gene is included in the reaction product is arranged. Examples of such probe placement regions are DNA chips and hybridizing regions.

  An example of a structure that holds and displaces the dispensing tip is a structure that holds and displaces the dispensing tip with a flexible material that is airtight, such as a diaphragm or a film.

  Other examples of the structure that holds the dispensing tip and supports it in a movable manner include a cover body in which the cover is integrated with the reaction plate, and an upper portion on the surface side of the reaction plate that is hermetically sealed with a sealing material. And a cover plate that is slidably held in a horizontal plane, and the dispensing tip is held on the cover plate so as to be slidable in the vertical direction while being airtight by another sealing material It is.

The reaction kit of the present invention is used for measurement of various reactions including chemical reactions and biochemical reactions.
Examples of the sample measured using the reaction kit of the present invention include various substances such as chemical substances, biological samples, and biological samples, and are not particularly limited.

  The reaction kit of the present invention is used in a state where the space on the surface side of the reaction plate is covered with a cover, and the sample introduction port can be penetrated by a sharp dispensing instrument, and dispensing after the penetration is performed. Since it is made of an elastic member that can close the through hole with elasticity when the instrument is pulled out, it can prevent foreign substances from entering the sample from the outside, and the reaction product can contaminate the external environment. The sample inlet can be easily sealed, and when there is a small amount of sample, there is no risk of drying, so that the analysis can be performed accurately.

  If the sample introduction part is formed as a container, the side of the container is the sample introduction port, and the upper part of the container is the opening for storing the liquid, so that from sample introduction to dispensing can be performed easily Become.

By attaching a cover film to the opening of the sample introduction part, it is possible to prevent the liquid from drying in the reaction kit and to prevent contamination with other reagents, thus enabling accurate analysis. .
If a seal film is affixed to the sample introduction port, it is possible to prevent the sample adhering to the elastic member from flowing out and being contaminated.

  If the reagent used for the sample reaction is introduced from the sample introduction part together with the sample, the versatility of the reaction kit is increased. On the other hand, if the reagent is stored in the reaction plate in advance, it is not necessary to prepare the reagent on the side of the apparatus for processing this reaction kit, so that the processing apparatus can be simplified.

If the dispensing tip is provided with a syringe operated from the outside of the cover, it is not necessary to provide a nozzle mechanism separately.
When the reaction plate is further equipped with a gene amplification part, even a sample containing only a very small amount of the gene to be measured can be amplified by a gene amplification reaction such as PCR or LAMP to increase the analysis accuracy It becomes like this.

If the dispensing tip is equipped with a filter inside the tip, even if the dispensing tip is not equipped with a syringe, foreign substances can be prevented from entering from the outside through the dispensing tip, It is also possible to prevent the reaction product from contaminating the external environment through the dispensing tip.
When a gene amplification reaction is performed, there is a problem that other DNA or the like enters the sample from the outside. There is also the problem that the amplified gene contaminates other samples. In the present invention, the gene amplification reaction is also performed in a closed space, and after the analysis is completed, it is discarded while being closed in the space, so that contamination from the outside can be prevented and there is no possibility of contaminating other samples. .

  If analysis of the reaction product in the reaction vessel is performed in the reaction vessel, an electrophoresis part provided at a different place from the reaction vessel, a probe arrangement region that reacts with the gene, etc., The types of samples handled can be expanded.

  A structure that holds the dispensing tip and supports it in a movable manner is realized by an airtight and flexible material, or the cover is composed of a cover body and a cover plate, and the dispensing tip is attached to the cover body. If it is supported so as to be movable by sliding and sliding of the dispensing tip with respect to the cover plate, a structure that holds the dispensing tip and supports it in a movable manner can be realized with a simple configuration.

FIG. 1 shows a reaction kit of one embodiment, (A) is a vertical sectional view, (B) is a plan view showing a reaction plate and a dispensing tip 20, and FIG. 2 is a perspective view of the embodiment. FIG.
As shown in FIG. 1, the reaction plate 2 includes a reaction container 4 for causing a sample to react on the surface side of a substrate 3 and a reagent container 12 containing a reagent used for the reaction of the sample and sealed with a film 14. I have.

  The reaction vessel 4 is provided as a recess on the surface of the substrate 3. When the temperature of the reaction vessel 4 is controlled from the outside during the reaction, it is preferable that the thickness of the reaction vessel 4 in that portion is thin in order to improve the thermal conductivity.

  The reagent container 12 is composed of a plurality of recesses formed in the substrate 3. Necessary reagents are accommodated in these recesses and covered with a film 14 that can be penetrated by a dispensing tip 20 described later. The film 14 is, for example, an aluminum foil, a laminated film of aluminum and a resin film such as a PET (polyethylene terephthalate) film, and is attached by fusion or adhesion so that it is not easily peeled off.

  A mixing portion for mixing the sample and the reagent may be formed as a concave portion on the surface of the substrate 3 as necessary, and such a mixing portion is covered with the film 14 in an empty state. It can be.

  In order to detect the reaction product in the reaction vessel 4, the reaction vessel 4 itself can be used as a detection unit by means such as irradiating the reaction vessel 4 with light from the outside. In addition, the detection unit can be provided independently of the reaction vessel 4. As such an independent detection unit, for example, a reaction solution after the reaction between the sample and the reagent is dispensed by the dispensing chip 20, and reagents for detecting the state after the reaction are arranged in advance. Can be. Such a detection unit can also have its surface covered with a film that can be penetrated by the dispensing tip 20. Similarly to the film 14, such a film can also be made of, for example, an aluminum foil, a laminated film of aluminum and a resin film such as a PET film, and can be attached by fusion or adhesion so that it is not easily peeled off. .

  Although the material of the board | substrate 3 containing the reaction container 4 is not specifically limited, Since this reaction kit is disposable, it is preferable that there exists a material which can be obtained cheaply. As such a material, for example, a resin material such as polypropylene and polycarbonate is preferable. When detection is carried out by absorbance, fluorescence, chemiluminescence or bioluminescence in the reaction vessel 4 or a separately provided detector, it is formed of a light transmissive resin so that optical detection can be performed from the bottom side. It is preferable. In particular, when fluorescence detection is performed, the substrate 3 may be formed of a material such as a resin having a low autofluorescence property (a property of generating less fluorescence from itself) and a light transmitting resin, such as polycarbonate. preferable. The thickness of the substrate 2 is 0.3 to 4 mm, preferably 1 to 2 mm. From the viewpoint of low autofluorescence for fluorescence detection, the substrate 3 is preferably thinner.

  A dispensing tip 20 is disposed on the upper surface of the reaction plate 2. In the case where the sample and the reagent or the reaction plate 2 is provided with an independent detection unit, the dispensing chip 20 further dispenses the reaction solution after the reaction to the detection unit. The dispensing tip 20 includes a syringe 22, and the dispensing operation is performed by driving the syringe 22 from the outside of the cover 24.

  As shown in FIG. 1C, the dispensing tip 20 may include a filter 23 inside instead of the syringe 22. The filter adsorbs foreign matter entering from the outside and prevents foreign matter from entering the space covered with the cover 24, and discharges reactants and reaction products from the space covered with the cover 24 to the outside. It is more effective in preventing it from being done.

  The cover 24 is provided so as to cover the upper space on the surface side of the reaction plate 2. The cover 24 includes a cover main body 26 that covers the peripheral portion and a bellows film 28 that covers the upper portion, and blocks the space on the surface side of the reaction plate 2 from the outside. The lower end of the cover main body 26 is fixed to the reaction plate 2 or is assembled integrally with the reaction plate 2 via a sealing material, and the shape of the cover 24 is maintained with rigidity. The bellows film 28 is made of a flexible diaphragm or a flexible film, and the dispensing tip 20 is disposed inside the space where the tip end portion is covered with the cover 24 and outside the space where the base end portion is covered with the cover 24. So that it is movable.

  The material of the cover 24 is not particularly limited as long as it can cover the upper space on the surface side of the reaction plate 2 in an airtight manner. However, since the reaction kit is disposable, it is inexpensive. It is preferable that there is a material that can be obtained. As such a material, for example, a resin material such as polypropylene or polycarbonate is preferable for the cover body 26, and nylon (registered trademark), polyvinyl chloride, silicone rubber, or other rubber materials are preferable for the bellows film 28.

  A part of the cover body 26 or the substrate 3 is provided with a holding member 30 for holding the dispensing tips 20 before and after use. The dispensing tips 20 are removed from the holding member 30 during dispensing. The upper part on the surface side of the reaction plate 2 can be freely moved.

Next, the sample introduction unit will be described.
A part of the cover body 26 is provided with a sample introduction part 32b for injecting a sample into the reaction kit from the outside via a sample introduction port 33b. The sample introduction port 33b can be penetrated by a sharp dispensing device 20b for sample injection, for example, a dispensing chip attached to the tip of a pipetter, and when the dispensing device 20b after the penetration is pulled out, the through hole is elastically formed. It is sealed by an elastic member 33c that can be closed by Therefore, the sample introduction port 33b maintains a sealed state even when the dispensing device 20b is penetrating and after being pulled out.
The sample introduction port 33b is a plate-like member having a tapered hole extending outward. The elastic member 33c is, for example, a rubber septum, and is sandwiched and fixed between the plate-like member provided with the sample introduction port 33b and the sample introduction portion 32b.

  In the case where the dispensing tip 20b is used with a dispensing tip attached to the tip, the dispensing device 20b includes the dispensing tip as a concept of the dispensing device. Therefore, in that case, the elastic member 33c of the sample introduction port 33b can be penetrated by the dispensing tip.

  The sample introduction part 32b forms a sample container 32, the side surface of the container 32 becomes a sample introduction port 33b, and the upper part of the container 32 is for dispensing the injected sample to a predetermined place on the sample plate. It has an opening. A cover film 14 a is attached to the opening of the container 32. A sample pretreatment liquid or reagent is sealed in the sample container 32 in advance.

By sticking the cover film 14a, it is possible to prevent the sample pretreatment liquid and the reagent in the sample container 32 from spilling during the movement and storage of the reaction kit.
As the cover film 14a, the same aluminum film as the film 14 can be attached.

  After the sample is introduced from the sample introduction port 33b, as shown in FIG. 3, the sample introduction port 33b can be sealed with the seal film 14b. Thereby, it can prevent that the sample (for example, blood etc.) etc. which adhered to the elastic member 33c flow out outside, and are contaminated.

  A specific example of the seal film 14b is obtained by applying an adhesive to a base material. As the substrate, polyethylene film, polypropylene film, polystyrene film, synthetic paper, polyimide film, variable information film, and the like can be used. As the adhesive applied to the substrate, PVA emulsion, SBR emulsion, acrylic emulsion, synthetic rubber emulsion, pressure sensitive adhesive, heat sensitive adhesive and the like can be used.

The seal film 14b may be attached to the cover body 26 in advance, and then peeled off at the time of sample injection, and may be attached to the cover body 26 again after the sample injection to seal the sample introduction port 33b. In that case, the adhesive applied to the base material of the seal film 14b is preferably an adhesive that can be easily peeled off.
As another form, the seal film 14b is not attached to the cover body 26 before sample injection, and is prepared separately by attaching a release paper to the seal film 14b so that it can be easily peeled off. The release paper may be peeled off and the seal film 14b may be attached to the cover body 26 to seal the sample introduction port 33b.

  This reaction kit is disposable, and after analyzing one sample, the entire reaction kit is discarded while the reaction plate 2 is covered with the cover 24.

Next, the operation | movement which analyzes a sample with the reaction kit of this Example is demonstrated.
Prior to the analysis, the sample is injected into the container 32 from the sample introduction port 33b through the elastic member 33c of the sample introduction port 33b by the dispensing device 20b. Thereafter, when the dispensing device 20b is pulled out, the sample introduction port 33b is closed by the restoring force of the elastic member 33c.

FIG. 3 is a view showing a reaction kit after sample introduction, (A) is a schematic perspective view, and (B) is a vertical sectional view.
The sample is introduced into the reaction kit, the upper opening is sealed with the film 14b, and the side surface (sample introduction port 33b) of the container 32 is sealed with an elastic member.

FIG. 4 shows a state in which the driving unit 36 starts engagement between the dispensing tip 20 and the syringe 22 with the sample introduced.
First, as shown in FIG. 5, the plunger holder 36 b, which is a syringe drive unit, descends and engages with the plunger of the syringe 22.
Subsequently, as shown in FIG. 6, the tip holder 36 a is also lowered and press-fitted into the dispensing tip 20 to hold the dispensing tip 20.

  Next, as shown in FIG. 7, the dispensing tip 20 is removed from the holding unit 30. Thus, the dispensing tip 20 can freely move in a state where it is blocked from the outside by the bellows film 28.

The dispensing tip 20 is moved to the sample in the sample container 32, penetrates the cover film 14a, injects the sample, and dispenses it into the reaction container 4.
Subsequently, the dispensing tip 20 is moved to the reagent container 12, penetrates the film 14, dispenses the reagent from the reagent container 12 to the reaction container 4, and is used for the reaction. During this reaction, the reaction vessel 4 is brought into contact with an external heat source as necessary, and is controlled to a predetermined temperature.

  During or after the reaction, the reaction product is detected. Here, it is assumed that the reaction product is optically detected from the outside of the reaction plate 2 in a state in the reaction vessel 4. Therefore, a detection unit is disposed below the reaction vessel 4 and detection is performed by optical or other means.

  In the above embodiment, the reaction plate 2 includes the reagent container 12, but the reaction plate 2 may not include the reagent container 12. In that case, the reagent can be used so as to be injected into the sample container 32 together with the sample and introduced into the reaction kit, or to be introduced into the reaction kit in another container not shown. .

FIG. 8 to FIG. 10 show examples of detection units used for detection of reaction products in the reaction container in the reaction kit of the present invention.
FIG. 8 shows an example of a detection unit comprising an absorbance detector. In this case, it is preferable that the reaction vessel 4 includes a pair of planes parallel to each other as an incident surface and an output surface of the measurement light.

  The detection unit 38a includes a light source 40a as an irradiation optical system, a pair of lenses 42a for condensing the light from the light source 40a, collimating the light into the reaction container 4, and irradiating the light to the reaction container 4, and a pair of lenses. A filter 44a, which is arranged in a portion made parallel light between 42a and selects light of a predetermined wavelength from light from the light source 40a as measurement light, and a mirror 46 which guides the measurement light to the incident surface of the reaction vessel 4 It is arranged on the optical path. As the light source 40a, in addition to a lamp light source such as a tungsten lamp that generates light having a wavelength in the ultraviolet region to the visible region, a light emitting diode (LED), a laser diode (LD), or the like is used. In addition, as a light receiving optical system, a photodetector 48a, a mirror 50 for guiding the light exiting the exit surface of the reaction vessel 4 to the photodetector 48a, and the light once converted into parallel light and then condensed to the photodetector 48a. A pair of lenses 52 to be incident on the filter 52 and a filter 54 a that is arranged in a portion where the parallel light is made between the pair of lenses 52 and selects a predetermined wavelength suitable for measurement are arranged on the optical path.

The reason why the lenses 42a and 52a make the respective lights once parallel is to improve the accuracy of wavelength selection in the filters 44a and 54a.
In this detection unit 38a, a wavelength suitable for detecting the reaction product is selected from the light from the light source 40a by the filters 44a and 54a, and the absorbance at that wavelength is measured to detect the reaction product.

FIG. 9 shows an example of a detection unit comprising a fluorescence detector.
This detection unit 38b collects the light from the light source 40b as an excitation optical system, and collimates the light from the light source 40b into a parallel light, and then condenses and irradiates the reaction vessel 4 with a pair of lenses 42b and the lens 42b. And a filter 44b that is arranged in the light path of the light beam and selects a predetermined excitation light wavelength from the light from the light source. In addition, a photodetector 48b as a light receiving optical system, and a pair of lenses 52b that receive fluorescence emitted from the reaction vessel 4 and convert it into parallel light, and then collect the light to enter the detector 48b, are parallel by the lens 52b. And a filter 54b that is arranged in the optical path of the fluorescent light and selects a predetermined fluorescence wavelength. Also in this case, the reason why the respective lights are once converted into parallel light by the lenses 42b and 52b is to increase the accuracy of wavelength selection in the filters 44b and 54b.

  In this detection unit 38b, the wavelength of the excitation light for exciting the reaction product by the filter 44b is selected from the light from the light source 40b and irradiated to the reaction product in the reaction vessel 4, and the fluorescence generated from the reaction product is emitted. Light is received by the light receiving optical system, a predetermined fluorescence wavelength is selected by the filter 54b, and the fluorescence is detected by the photodetector 48b.

FIG. 10 is an example of a detection unit for detecting chemiluminescence or bioluminescence from the reaction product.
The detection unit 38c includes a light detector 48c for detecting light emitted from the reaction vessel 4, a lens 52c for receiving light emitted from the reaction vessel 4 and guiding it to the light detector 48c, and collected light. Is provided with a filter 54c for selecting a predetermined emission wavelength.
In the detection unit 38c, the light from the chemiluminescence or bioluminescence from the reaction product in the reaction vessel 4 is collected by the lens 52c, the wavelength is selected by the filter 54c, and is detected by the photodetector 48c.

  FIGS. 11 to 15 show other embodiments in which the structure of the reaction plate is different. In the reaction plate of the above embodiment, the reaction product is detected by the reaction vessel 4, but in the embodiment shown in FIGS. 11 to 15, the reaction plate further includes an analysis unit for analyzing the reaction product. ing.

  The reaction plate 2a in the embodiment of FIG. 11 includes an electrophoresis unit as an analysis unit. An example of the electrophoresis unit is an electrophoresis chip 100, which includes a reaction product injection unit 103, an electrophoresis separation channel 102, and electrophoresis voltage application electrodes 106a to 106d. Here, in addition to the electrophoresis separation channel 102, a sample introduction channel 104 that crosses the electrophoresis separation channel 102 and introduces the sample into the electrophoresis separation channel 102 is also provided. Alternatively, the sample may be configured to be directly introduced into one end of the electrophoresis separation channel 102. In order to detect fluorescence from the back side, the electrophoresis chip 100 is formed of a material such as glass or quartz, such as low autofluorescent and light-transmitting resin, such as polycarbonate.

  The reaction plate 2 a also includes a separation buffer solution container 15 that is sealed with a film that contains the separation buffer solution injected into the flow paths 102 and 104 and can be inserted at the tip of the dispensing tip 20 on the surface side. .

Electrophoresis voltage application electrodes 106a to 106d are connected to the ends of the flow paths 102 and 104, respectively, and are led to the outside of the cover 24 so that they can be connected to a power supply device provided outside the reaction kit.
Reservoirs are provided at the ends of the flow paths 102 and 104, and the separation buffer solution accommodated in the separation buffer solution container 15 is placed in these reservoirs.
An example of the use of this example for gene analysis will be described. The reagent container 12 contains a PCR reaction reagent. The reaction container 4 becomes a PCR reaction container.

When measuring a gene sample with the reaction kit of this embodiment, the sample is introduced into the sample container 32 in the reaction kit from the sample introduction port 33b by the dispensing device 20b. In the processing apparatus, the dispensing tip 20 dispenses the sample container 32 to the reaction container 4, and the dispensing tip 20 dispenses the PCR reaction reagent from the reagent container 12 to the reaction container 4. After superposing mineral oil (not shown), the reaction solution in the reaction vessel 4 is controlled to have a predetermined temperature cycle to cause a PCR reaction.
In the electrophoresis chip 100, the dispensing buffer 20 supplies the separation buffer solution from the separation buffer solution container 15 to the flow paths 102 and 104 through the reservoir of the electrophoresis chip 100.

The reaction solution after completion of the PCR reaction is injected as a sample from the reaction vessel 4 into the injection portion 103 of the electrophoresis chip 100 where the separation buffer solution is supplied. Thereafter, a voltage is applied to the channels 102 and 104 by the electrodes 106a to 106d from the power supply device 101 (see FIG. 12) provided in the processing apparatus to introduce the sample into the electrophoresis separation channel 102, and then the electricity is supplied. The electrophoresis separation channel 102 is migrated and separated.
In order to detect the sample component separated by electrophoresis, the processing apparatus is provided with a detection unit 38d.
Here, the reaction vessel 4 is used as a PCR reaction vessel, but a PCR reaction vessel may be provided separately from the reaction vessel 4.

  The detection unit 38d is shown in FIG. The detection unit 38d includes an excitation optical system and a fluorescence light receiving optical system, and performs fluorescence detection of a sample component passing through a predetermined position of the electrophoresis separation channel 102. Since the detection unit 38d detects the fluorescence of the sample component passing through the fixed position, the detection unit 38d does not need to be moved.

  The excitation optical system includes a light source 40c, a lens 42c that collects the light from the light source 40c to make parallel light, and is arranged in the optical path of the light beam that has been made parallel light by the lens 42c. And a filter 44c for selecting.

  In order to irradiate the excitation light from the excitation optical system to a predetermined position of the electrophoresis separation channel 102 from the back surface of the electrophoresis chip 100, and to receive the fluorescence generated from the position into parallel light, An objective lens 55 is provided. The dichroic mirror 53 has a spectral wavelength set so as to reflect the light having the excitation light wavelength used in this embodiment and to transmit the light having the fluorescence wavelength.

  The fluorescence light receiving optical system is disposed at a position for receiving fluorescence that has been converted into parallel light by the objective lens 55 and transmitted through the dichroic mirror 53, and a filter 54c that selects a predetermined fluorescence wavelength from the fluorescence transmitted through the dichroic mirror 53; A lens 52c that collects the fluorescence selected by the filter 54c and enters the detector 48c. Also in this case, the reason why the respective lights are once converted into parallel light by the lenses 42c and 55 is to increase the accuracy of wavelength selection in the filters 44c and 54c.

  In this detection unit 38d, the wavelength of the excitation light for exciting the reaction product by the filter 44c is selected from the light from the light source 40c, and the reaction product passing through a predetermined position of the electrophoresis separation channel 102 is irradiated. Then, the fluorescence generated from the reaction product is received by the light receiving optical system, a predetermined fluorescence wavelength is selected by the filter 54c, and the fluorescence is detected by the photodetector 48c.

  The reaction plate 2b in the embodiment of FIG. 13 includes a DNA chip 110 as an analysis unit. When a gene is included in the reaction product, a probe that reacts with the gene is fixed to the DNA chip 110. The DNA chip 110 is formed of a low autofluorescent and light-transmitting resin such as polycarbonate or glass in order to detect fluorescence from the back side.

The reaction plate 2a accommodates a washing solution for separating and removing the reaction product that has not been bound from the reaction product bound to the probe in the DNA chip 110 on the surface side, and can be inserted at the tip of the dispensing tip 20 A cleaning liquid container 17 sealed with a transparent film is also provided.
An example of the use of this example for gene analysis will be described. The reagent container 12 contains a PCR reaction reagent. The reaction container 4 becomes a PCR reaction container.

  When measuring a gene sample with the reaction kit of this embodiment, the sample is introduced into the sample container 32 in the reaction kit from the sample introduction port 33b by the dispensing device 20b. In the processing apparatus, the dispensing tip 20 dispenses the sample container 32 to the reaction container 4, and the dispensing tip 20 dispenses the PCR reaction reagent from the reagent container 12 to the reaction container 4. After superposing mineral oil (not shown), the reaction solution in the reaction vessel 4 is controlled to have a predetermined temperature cycle to cause a PCR reaction.

  The reaction solution after completion of the PCR reaction is injected as a sample from the reaction vessel 4 into the DNA chip 110 by the dispensing chip 20. After the incubation, the washing solution is injected from the washing solution container 17 into the DNA chip 110 by the dispensing tip 20, and the reaction product that is not bound to the probe is sucked and removed together with the washing solution by the dispensing tip 20.

By labeling the reaction product with a fluorescent substance, the reaction product bound to the probe can be detected by fluorescence. Thereby, it is detected that the gene corresponding to the probe at the position where the fluorescence was detected was included in the sample.
In order to detect the reaction product bound to the probe with the dispensing tip 20, a detection unit 38e is provided in the processing apparatus.

  The detection unit 38e is shown in FIG. The configuration of the optical system of the detection unit 38e is the same as that of the detection unit 38d shown in FIG. Since this detection unit 38e must move over the position of the probe arranged on the DNA chip 110, it is different from the detection unit 38d shown in FIG. As shown in FIG. 21, the movement can be realized by moving the table 82 in the X direction and moving the detection unit 38e in the Y direction.

  The reaction plate 2c in the embodiment of FIG. 15 includes a DNA chip 120 as an analysis unit. The DNA chip 120 is different from the DNA chip 110 of the embodiment of FIG. 13 in that the detection is performed not by fluorescence detection but electrically. A phenomenon is used in which the current value of the probe changes depending on whether or not the sample gene is bound to the probe. Since the DNA chip 120 does not perform optical detection, the DNA chip 120 does not need to be a light-transmitting material and may be insulative.

  When a gene is included in the reaction product, a probe that reacts with the gene is fixed to the DNA chip 120. From each of these probes, an electrode is taken out on the back side, and the current value of each flow is measured. In this example, the sample need not be labeled with a fluorescent material.

In order to perform the measurement with the DNA chip 120, the electrode taken out from each probe to the back side is connected to a detector 122 provided in the processing apparatus, and the current value of each probe is measured.
The reaction plate 2c also has a cleaning liquid for separating and removing unreacted reaction products from the reaction products bound to the probes in the DNA chip 120 on the surface side, and can be inserted at the tip of the dispensing chip 20 A cleaning liquid container 17 sealed with a transparent film is provided. The reagent container 12 stores a PCR reaction reagent. The reaction container 4 becomes a PCR reaction container.

  When measuring a gene sample with the reaction kit of this embodiment, the sample is introduced into the sample container 32 in the reaction kit from the sample introduction port 33b by the dispensing device 20b. In the processing apparatus, the dispensing tip 20 dispenses the sample container 32 to the reaction container 4, and the dispensing tip 20 dispenses the PCR reaction reagent from the reagent container 12 to the reaction container 4. After superposing mineral oil (not shown), the reaction solution in the reaction vessel 4 is controlled to have a predetermined temperature cycle to cause a PCR reaction.

  The reaction solution after completion of the PCR reaction is injected as a sample from the reaction container 4 into the DNA chip 120 by the dispensing chip 20. Thereafter, a cleaning solution is injected from the cleaning solution container 17 into the DNA chip 120 by the dispensing tip 20, and a reaction product that is not bound to the probe is sucked and removed together with the cleaning solution by the dispensing tip 20.

In order to detect the reaction product bound to the probe by the dispensing chip 20, the processing device is provided with a detector 122, which removes the reaction product that has not bound to the probe, and the detector 122 causes each probe to be detected. Measure the current value.
In the embodiment of FIG. 13 or FIG. 15, genes can be measured in the same manner even if the DNA chips 110 and 120 are replaced with hybridization regions.

  FIG. 16 shows another embodiment having a different cover structure. A part of the cover for supporting the dispensing tip 20 movably and covering the upper part of the reaction plate 2 is the bellows film 28 in the embodiment of FIG. 1, whereas it is flexible in the embodiment of FIG. The difference is that the film-shaped material 28a is deformed. As the film-like material 28a, like the bellows film 28, nylon (registered trademark), polyvinyl chloride, silicone rubber and other rubber materials are preferable.

Further, the sample container 32 in the embodiment of FIG. 16 is the same as that of the embodiment of FIG.
These detection units 38a, 38b, and 38c are arranged so as to be below the reaction plate 2 in the processing apparatus that performs the processing of the reaction kit, with the reaction kit mounted on the processing apparatus.

FIG. 17 shows still another example of the reaction kit. (A) is a vertical sectional view, (B) is a horizontal sectional view, and (C) is an external perspective view.
In this embodiment, the cover that supports the dispensing tip 20 so as to be movable is made of a rigid material. The cover body 60 of the cover 24 a has an opening 62 above the reaction plate 2, and a cover plate 64 for supporting the dispensing tip 20 movably within the range of the opening 62 is provided in the opening 62. . The cover body 60 has a double structure with a gap around the opening 62, and the cover plate 64 includes a sealing material 66 around the opening, and the sealing material 66 has a double structure around the opening 62 of the cover body 60. The cover plate 64 can be moved in the X direction within a horizontal plane by being moved in the X direction by being sandwiched between the gaps in the structure. The dispensing tip 20 is supported on the cover plate 64 via another sealing material 68 so as to be slidable in the vertical direction (Z direction).

  In this embodiment, the cover plate 64 moves in a horizontal plane while being kept airtight by the seal structure of the seal material 66 and the double structure gap above the cover body 60, and the dispensing tip 20 is airtight by the seal material 68. The dispensing tip 20 can freely move in the upper space of the reaction plate 2 in both the vertical and horizontal directions by moving in the vertical direction while maintaining

  FIG. 18 shows still another embodiment. Compared to the embodiment of FIG. 17, the cover plate 64 can move in both the X and Y directions, and is different in that the number of reagent containers 12 in the reaction plate 2 is increased, and other structures are the same. is there.

  FIG. 19 shows still another embodiment. This embodiment is different from the embodiment of FIG. 17 in that a cover plate 64a constituting an upper member of the cover is rotatably supported in the in-plane direction in order to move the dispensing tip 20 in the in-plane direction. The cover plate 64a has a disk shape, and a sealing material 66 is attached around the cover plate 64a. The sealing material 66 is supported by a gap having a double structure provided at the upper part of the cover body 60, and supports the cover plate 64a so as to be rotatable while maintaining airtightness. The dispensing tip 20 is supported by the cover plate 64a so as to be movable in the vertical direction by the sealant 68, and the supported position is a position away from the rotation center of the cover plate 64a.

  As the cover plate 64a rotates, the position of the dispensing tip 20 moves on the circumference around the rotation center of the cover plate 64a. In the reaction plate 2, the arrangement is determined so that the reaction container 4, the reagent container 12, and the sample container 32 are positioned on the movement trajectory of the dispensing tip 20.

  FIG. 20 shows still another embodiment. Compared with the embodiment of FIG. 19, the cover plate 64 a also has an opening 70, and the periphery of the opening 70 has a double structure, and the other cover plate 71 moves to the gap of the double structure through the sealing material 72. Supported as possible. The dispensing tip 20 is supported on the cover plate 71 by another sealing material 68 so as to be movable in the vertical direction.

  The dispensing tip 20 can be moved in the in-plane direction by the sealing material 72. Therefore, the movement range of the dispensing tip 20 is a donut centered on the rotation center of the cover plate 64a due to both the circumference due to the rotation of the cover plate 64a and the movement range in the horizontal plane where the small cover plate 71 can be moved by the sealing material 72. The range of the shape can be moved. In this way, the movement range of the dispensing tip 20 is widened, so that the number of reaction containers 4 and reagent containers 12 arranged in the movement range can be increased, and freedom for the arrangement of these containers including the sample container 32 can be increased. The degree increases.

FIG. 21 is a perspective view schematically showing the inside of an example of a processing apparatus for processing the reaction kit according to the present invention.
80 represents the reaction kit shown in the above examples. The reaction kit 80 is mounted on a table 82 which is a reaction kit mounting portion. The table 82 has an opening on the lower surface side of the reaction kit 80, and a detection unit 38 for optically detecting the reaction product in the reaction container 4 of the reaction kit 82 is disposed below the table 82. A temperature control (temperature control) unit 83 that controls the temperature of the reaction kit 82 is also arranged on the table 82. When the gene amplification reaction is performed using the reaction container 4 of the reaction kit or a separately provided gene amplification reaction container, the temperature control unit 83 performs temperature control for the gene amplification reaction. When the reaction kit includes an analysis unit that requires temperature control, the temperature adjustment unit 83 controls the temperature of the analysis unit. The temperature control unit 83 includes those having both of these functions. The detection unit 38 is the one shown in FIGS. The table 82 moves in the front-rear direction (X direction), while the detection unit 38 is supported so as to move in the lateral direction (Y direction) orthogonal thereto.

  Near the table 82, a drive unit 36 for driving the dispensing tip 20 is mounted so as to be movable in the Y direction and the Z direction. As shown in FIG. 4, the drive unit 36 is engaged with the proximal end portion of the dispensing tip 20 to hold the dispensing tip 20, and the syringe provided on the dispensing tip 20. The syringe drive part 36b which engages with 22 plungers and drives a syringe is provided on the same axis | shaft, and both the movement of the dispensing tip 20 and the drive of the syringe 22 can be performed.

  FIG. 22 is a block diagram showing a control system in an example of a reaction kit processing apparatus. In order to control the processing operation for the reaction kit 80 mounted on the table 82, a control unit 84 including a dedicated computer (CPU) or a general-purpose personal computer is provided. The control unit 84 moves and dispenses the dispensing tip 20 by the drive unit 36 engaged with the base end of the dispensing tip 20, controls the temperature by the temperature control unit 83, and measures the light in the reaction container 4 of the reaction kit 80. Alternatively, the detection operation by the detection unit 38 that irradiates the excitation light and optically detects the reaction product is controlled.

  For example, a personal computer (PC) 86 may be connected to the control unit 84 as an external computer in order to use the control unit 84 as an input unit operated from the outside or as a monitor for displaying inspection results. .

  The present invention can be used for measurement of various chemical reactions and biochemical reactions.

It represents one Example of a reaction kit, (A) is a vertical sectional view, (B) is a plan view showing a reaction plate and a dispensing tip, and (C) is a schematic showing another example of a dispensing tip. It is sectional drawing. It is an external appearance perspective view of the same Example. The state after the sample introduction of the same Example is represented, (A) is an external appearance perspective view, (B) is a vertical sectional view. It is a vertical sectional view showing a state where a sample is introduced in the same example. It is a vertical sectional view which shows the state which the syringe drive part of the drive unit engaged with the plunger of the syringe in the Example. It is a vertical sectional view showing the state where the tip holding part of the drive unit is engaged with the dispensing tip in the same embodiment. It is a vertical sectional view which shows the state by which the dispensing tip was removed from the holding | maintenance part in the Example. It is a vertical sectional view showing a first example of a detection unit used for detecting a reaction product in the reaction kit of the present invention. It is a vertical sectional view showing a second example of a detection unit used for detection of a reaction product in the reaction kit of the present invention. It is a vertical sectional view showing a third example of a detection unit used for detecting a reaction product in the reaction kit of the present invention. It is a figure showing the other Example of the reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip. It is the vertical sectional view which shows the example of the detection unit used for the detection of the reaction product in the reaction kit of the Example with a reaction kit. It is a figure showing the further another Example of a reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip. It is the vertical sectional view which shows the example of the detection unit used for the detection of the reaction product in the reaction kit of the Example with a reaction kit. FIG. 10 is a vertical sectional view showing still another embodiment of the reaction kit together with an example of a detection unit used for detecting a reaction product. It is a vertical sectional view showing other examples of a reaction kit. It is a figure showing the further another Example of a reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip, (C) is an external appearance perspective view. It is a figure showing the further another Example of a reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip, (C) is an external appearance perspective view. It is a figure showing the further another Example of a reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip, (C) is an external appearance perspective view. It is a figure showing the further another Example of a reaction kit, (A) is a vertical sectional view, (B) is a top view which shows a reaction plate and a dispensing tip, (C) is an external appearance perspective view. It is an internal schematic perspective view which shows an example of a reaction kit processing apparatus. It is a block diagram which shows the control system in the reaction kit processing apparatus.

Explanation of symbols

2, 2a, 2b, 2c Reaction plate 3 Substrate 4 Reaction container 12 Reagent container 14, 14a, 14b Film 20 Dispensing tip 20b Dispensing instrument dispensing tip 22 Syringe plunger 23 Filter 24 Cover 26 Cover body 28 Bellows film 33b Sample introduction port 33c Elastic member 32 Sample container 32b Sample introduction part 64, 64a, 71 Cover plate 66, 68, 72 Sealing material 100, 110, 120 DNA chip 106 Electrode 102 Flow path for electrophoresis separation

Claims (14)

A reaction plate provided with a reaction vessel for causing the sample to react on the surface side;
A dispensing tip disposed above the surface side of the reaction plate;
A cover that covers the upper space on the surface side of the reaction plate, and supports the dispensing tip so that the tip is inside the space and the base is outside.
A sample introduction part for injecting a sample into the space from the outside via a sample introduction port provided in a sealable manner in a part of the cover;
The reaction is characterized in that the sample introduction port is constituted by an elastic member that can be penetrated by a sharp dispensing device with a sharp tip, and that the through hole can be closed by elasticity when the dispensing device after the penetration is pulled out. kit.   The sample introduction part forms a container, the sample introduction port is a side surface of the container, and has an opening at the top of the container, and the sample pretreatment liquid or reagent is sealed in advance in the container The reaction kit according to claim 1.   The reaction kit according to claim 1, wherein the opening is sealed with a cover film attached thereto.   The reaction kit according to any one of claims 1 to 3, wherein the sample introduction port is hermetically sealed by attaching a seal film to the sample introduction port.   The reaction kit according to any one of claims 1 to 4, wherein the reaction plate further includes a reagent container which contains a reagent used for sample reaction and is sealed with a film.   The reaction kit according to any one of claims 1 to 5, wherein the dispensing tip includes a syringe operated from the outside of the cover, and a dispensing operation is performed by operating the syringe.   The reaction kit according to any one of claims 1 to 6, wherein the dispensing tip includes a filter in a tip portion.   The reaction kit according to any one of claims 1 to 7, wherein the reaction plate includes a gene amplification unit that performs a gene amplification reaction on a surface side thereof.   The reaction kit according to any one of claims 1 to 8, wherein the reaction container is made of a light-transmitting material so that optical measurement is possible from the bottom.   The reaction kit according to any one of claims 1 to 9, wherein the reaction plate further includes an analysis unit for analyzing a reaction product in the reaction vessel on a surface side thereof.   The reaction kit according to claim 10, wherein the analysis unit is an electrophoresis unit that performs electrophoretic separation of reaction products.   The reaction kit according to claim 10, wherein the analysis unit is a region where a probe that reacts with a gene when the reaction product contains a gene is arranged.   The reaction kit according to any one of claims 1 to 12, wherein the cover holds the dispensing tip and is movably supported by an airtight and flexible material. The cover is disposed on a cover main body integrated with the reaction plate and on the upper surface side of the reaction plate, and is kept slidable in a horizontal plane while being airtight with a sealant with respect to the cover main body. A cover plate,
The reaction kit according to any one of claims 1 to 12, wherein the dispensing tip is held on the cover plate so as to be slidable in a vertical direction while being airtight with another sealing material.

Images