JP2007322291A - Dispensing tip and reaction kit using same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To prevent an outside and a dispensing solution from being contaminated via aerosol in a dispensing tip. <P>SOLUTION: This dispensing tip 20 of the present invention includes a dispensing nozzle 19 provided in a tip part, a syringe 21 connected to an upper part of the dispensing nozzle 19 and having a hollow inside, and a plunger 22 sliding vertically inside the syringe 21 to suck and deliver a liquid in the dispensing nozzle 19. The dispensing tip 20 includes a separation member having air tightness for separating an inside of the nozzle 19 from the outside and flexibility capable of sliding the plunger 22, between an upper part of the plunger 22 and an upper part of the syringe 21. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a dispensing chip suitable for performing various types of analysis and analysis in the field of biological analysis, biochemical analysis, or chemical analysis in general, and a reaction kit using the same. is there.

  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 addition, a hollow conical tip having a sharp tip is used as a dispensing tip that is attached to the tip of a suction / discharge nozzle for dispensing a solution such as a sample or a reagent in these reactors. .

However, when dispensing a solution with a dispensing tip, solution components and reaction products leak from the dispensing tip to the suction / discharge nozzle side via aerosol, causing contamination and contamination to the outside. Sometimes. Conversely, the solution to be dispensed may be contaminated via the aerosol inside the dispensing tip.
Therefore, an object of the present invention is to prevent contamination of the outside or the solution to be dispensed through the aerosol in the dispensing tip.

  The dispensing tip of the present invention is connected to the dispensing nozzle provided at the tip, the upper part of the dispensing nozzle, the inside of the syringe is hollow, and the dispensing nozzle slides up and down in the syringe. A plunger for sucking and discharging the liquid is provided. Between the upper part of the plunger and the upper part of the syringe, there is provided an isolation member having airtightness separating the inside of the nozzle from the outside and flexibility for sliding the plunger. .

  When the plunger is pulled up, the loop space formed by the plunger, the syringe, and the isolation member between the upper part of the plunger and the upper part of the syringe is decompressed, and the plunger and the isolation member may not move smoothly. Therefore, you may make it provide the air hole which leads to the loop-shaped space formed with a syringe, a plunger, and a separating member.

  When using a conventional microwell reaction plate, the top surface of the reaction plate is open to the atmosphere, and foreign substances may enter the sample from the outside. Conversely, reaction products may contaminate the external environment. It is possible. Therefore, the reaction kit of the present invention includes a reaction plate provided with a reaction vessel for causing a sample to react on the surface side, and a surface of the reaction plate in order to prevent entry of foreign substances from the outside of the reaction plate and environmental contamination to the outside. The upper part of the dispensing tip of the present invention disposed on the side of the reaction plate and the upper plate space on the surface side of the reaction plate are covered, and the tip of the dispensing tip is located inside the space above the plate and the base end is located outside. And a cover that is movably supported.

  In order to prevent decompression of the loop-shaped space generated when the plunger is pulled up, it is convenient and preferable to provide an air hole. However, when the air hole is provided outside the cover, the loop-shaped space is separated from the outside by an isolation member. Since the effect of sealing is diminished, an air hole is provided in a portion of the base end portion of the syringe that is disposed inside the space above the plate, and the space above the plate covered with air in the loop space is covered with a cover It is preferable to exchange it with the inside.

  In using this reaction kit, the sample must be introduced into the space on the plate covered with the cover by some method. The introduction method is not particularly limited. For example, a sample introduction unit for injecting a sample into the space on the plate from the outside through an opening provided in a sealable part of the cover is further provided. May be.

  The reagent used for the reaction of the sample must also be introduced into the space on the plate covered with the cover by any method, and the method is not particularly limited. For example, the reagent is 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 which 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 upper surface of the plate on the surface of the reaction plate is covered with a cover and shielded from the outside, and the reaction for the sample is performed in the upper space of the plate. 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.

  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 by a flexible material having airtightness, 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.

  Since the dispensing tip of the present invention is provided with an isolation member having an airtightness that separates the inside of the dispensing nozzle from the outside between the upper part of the plunger and the upper part of the syringe and a flexibility that allows the plunger to slide, the plunger is slid. Even if it is moved, it is possible to suppress leakage of the components and reaction products of the solution to the outside from the isolation member portion of the plunger and the syringe through the aerosol, and as a result, contamination and contamination can be prevented.

  In addition, if an air hole is provided to the loop-shaped space formed by the syringe, the plunger, and the separating member, it is possible to prevent the loop-shaped space from being depressurized when the plunger is pulled up. The isolation member can move smoothly, and an accurate analysis can be performed.

  Since the reaction kit of the present invention is used in a state where the space on the plate on the surface side of the reaction plate is covered with a cover, foreign substances can be prevented from entering the sample from the outside, and the reaction product can be prevented. Contamination of the external environment can be prevented, and the dispensing tip of the present invention can completely close the inside of the dispensing nozzle with the external space with a simple mechanism, so that the reaction vessel can be made compact. it can.

  If an air hole is provided in a portion of the base end portion of the syringe that is disposed inside the space above the plate, the air existing in the looped space of the syringe in a state of being completely closed with the external space is allowed to be It becomes possible to exchange with the inside, and it becomes possible to prevent external contamination, downsize the apparatus, and improve the accuracy of analysis.

  When the sample introduction unit is further provided, the sample introduction operation into the space on the plate covered with the cover is facilitated.

  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 carried out in a closed space on the plate. After the analysis is completed, the sample is disposed in the closed space on the plate, so that contamination from the outside can be prevented and other samples can be contaminated. There is no risk of doing so.

  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, and 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, aluminum foil, a laminated film of aluminum and a resin film such as PET (polyethylene terephthalate) film, and is attached by fusion or adhesion so that it does not easily peel 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. Similar 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, or the like, and can be attached by fusion or adhesion so as not to be 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 21, and the dispensing operation is performed by driving the syringe 21 from the outside of the cover 24.

Next, the dispensing tip 20 and the catch mechanism 90 for mounting it will be described.
3A and 3B show the dispensing tip 20 and a catch mechanism for mounting the dispensing tip 20A. FIG. 3A shows a state before the catch mechanism is attached to the dispensing tip, and FIG. 3B shows that the catch mechanism is attached to the dispensing tip. (C) shows a cross-sectional view of a state in which the plunger 22 in the dispensing tip is pulled up.
The dispensing tip 20 includes a dispensing nozzle 19 that sucks and discharges liquid from the tip, a syringe 21 that is connected to the upper part of the dispensing nozzle 19 and has a hollow inside, and slides up and down in the syringe 21. And a plunger 22 that sucks and discharges liquid by moving.

  The catch mechanism 90 is capable of separately holding the plunger 22 and the dispensing tip 20, and is provided on the outer periphery of the plunger holder 36b and the plunger holder 36b that are positioned coaxially with the plunger 22, A sleeve 93 provided so as to be relatively movable in the vertical direction (direction parallel to the axis) with respect to 36b, and relative to the sleeve 93 outside the sleeve 93 in the vertical direction (direction parallel to the axis). The chip holder 36a is provided so as to be movable.

Coil springs 92a and 94a are provided in a compressed state between the plunger holder 36b and the sleeve 93 and between the sleeve 93 and the tip holder 36a.
The distal ends of the plunger holder 36b and the tip holder 36a grip the plunger 22 and the dispensing tip 20 by friction or a stopper. For example, the concave and convex portions may be provided on the inner surface of the plunger holder 36b, and the outer surface of the plunger 22 may be shaped to fit the concave and convex portions.

  An isolation member 96 is formed so as to connect the packing 98 a provided on the upper portion of the plunger 22 and the packing 98 b provided on the upper portion of the syringe 21. The separating member 96 is preferably provided with airtightness and flexibility, and for example, a diaphragm or a thin film can be used. As a flexible material, silicone rubber, ethylene propylene rubber (EPDM), butyl rubber, or the like can be used.

A loop-shaped space 95 is formed by the separating member 96, the syringe 21 and the plunger 22.
The volume of the loop space 95 changes as the plunger 22 moves. An air hole is provided in a portion of the proximal end portion of the syringe 21 disposed inside the plate space so that air can be exchanged between the loop space 95 and the plate space covered by the bellows film 28. 99 is provided.

FIG. 4 is a vertical sectional view showing mounting operations (A1) to (A6) in which the catch mechanism 90 mounts the dispensing tip 20, and releasing operations (B1) to (B2) in which the catch mechanism 90 releases the dispensing tip 20. It is.
(A 1) is an initial state before the catch mechanism 90 mounts the dispensing tip 20, and the plunger holder 36 b is accommodated in the sleeve 93. The tip of the plunger holder 36 b is provided with a notch like the tip of a mechanical pencil, and the outer diameter of the tip is widened when it comes out of the sleeve 93.

(A2) is a state when the plunger holder 36b is lowered, and the tip of the plunger holder 36b that has come out from the inside of the sleeve 93 expands to a size sufficient to hold the plunger 22. The coil spring 92a is contracted by the force caused by the lowering of the plunger holder 36b.
(A3) is a state when the plunger holder 36b whose tip is widened is temporarily attached to the plunger 22. Since both the coil springs 92a and 94a are contracted, the tip holder 36a is also lowered at the same time and is in contact with the syringe 21 to prepare for mounting.

(A4) is a state in which the plunger holder 36b temporarily attached with the plunger 22 is pulled up, and the coil spring 92a is extended thereby.
(A5) is a state when the plunger holder 36b and the tip holder 36a are lowered simultaneously, and the plunger holder 36b is fully attached to the plunger 22 and the tip holder 36a is fully attached to the syringe 21.
(A6) shows a state in which the plunger 22 is slid in the vertical direction after the main mounting.

(B1) is a state in which the catch mechanism 90 is mounted with the dispensing tip 20.
(B2) shows a state in which the dispensing tip 20 is released from the catch mechanism 90, and a downward force is applied to the plunger holder 36b and a lifting force is applied to the tip holder 36a.
The catch mechanism 90 releases the dispensing tip 20 by expanding the tip of the plunger holder 36b to a size that releases the plunger 22.

Next, the cover 24 will be described.
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 located 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 materials, 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.

In order to introduce the sample into the reaction plate 2 from the outside of the cover 24, an opening 31 is provided in a part of the cover body 26, and a sample container 32 is attached to the opening 31 so as to be openable and closable. The sample container 32 is formed with a recess opened upward for injecting a sample. When a sample is injected into the recess and positioned inside the cover 24, an adhesive is placed inside the plate 34 so that the plate 34 holding the sample container 32 is in close contact with the cover body 26 and seals the opening 31. It is applied or is sandwiched between the cover main body 26 via a sealing material. Therefore, the opening 31 is an opening that can be sealed.
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 sample container 32 from the opening 31, and then the sample container 32 is fixed to the cover body 26 by closing the opening 31 by the sample container 32, and the sample is covered with the cover 24 of this reaction kit. It is shut off from the outside when it is introduced into the closed space.

FIG. 5 shows a state in which the drive unit 36 starts engagement between the dispensing tip 20 and the syringe 21 with the sample introduced.
First, as shown in FIG. 6, the plunger holder 36 b that is a syringe drive unit is lowered and engaged with the plunger 22 of the syringe 21.
Subsequently, as shown in FIG. 7, 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. 8, the dispensing tip 20 is removed from the holding unit 30. Thus, the dispensing tip 20 can be freely moved 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, and the sample is injected and dispensed 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. 9 to FIG. 11 show examples of the detection unit used for detecting the reaction product in the reaction container in the reaction kit of the present invention.
FIG. 9 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. 10 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. 11 shows 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.

  FIG. 12 to FIG. 16 show other embodiments having different reaction plate structures. 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. 12 to 16, the reaction plate further includes an analysis unit for analyzing the reaction product. ing.

  The reaction plate 2a in the embodiment of FIG. 12 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 contains a separation buffer solution injected into the flow paths 102 and 104 and is sealed with a film that 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 from the sample container 32, and the reaction kit is attached to the processing apparatus. 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. 13) provided in the processing apparatus, and the sample is introduced 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. 14 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 thin 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 from the sample container 32, and the reaction kit is attached to the processing apparatus. 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 has to move over the position of the probe arranged on the DNA chip 110, it is different from the detection unit 38d shown in FIG. The movement can be realized by movement of the table 82 in the X direction and movement of the detection unit 38e in the Y direction, as shown in FIG.

  The reaction plate 2c in the embodiment of FIG. 16 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. 14 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 thin 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 from the sample container 32, and the reaction kit is attached to the processing apparatus. 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. 14 or FIG. 16, genes can be measured in the same manner even if the DNA chips 110 and 120 are replaced with hybridization regions.

  FIG. 17 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, but is flexible in the embodiment of FIG. The difference is that the film-shaped material 28a is deformed. As the film-like material 28 a, nylon (registered trademark), polyvinyl chloride, silicone rubber, and other rubber materials are preferable as in the case of the bellows film 28.

  Further, in the embodiment of FIG. 1, one side of the sample container is rotatably supported by the cover body 26, whereas the sample container 32 a in the embodiment of FIG. 17 is slidably attached to the cover body 26. Is different. Also in such a sample container 32a, the sample container 32a can be dispensed to the sample container 32a by being pulled out from the cover body 26 to the outside. Further, an adhesive is applied to the inside of the plate 34 of the sample container 32a, and the opening 31 is sealed inside the plate 34 by pushing the sample container 32a into the inside of the cover main body 26, or the opening 31 is formed by a sealing material. The point which can be sealed 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. 18 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. 19 shows still another embodiment. Compared with the embodiment of FIG. 18, the cover plate 64 can be moved 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. is there.

  FIG. 20 shows still another embodiment. This embodiment is different from the embodiment of FIG. 18 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. 21 shows still another embodiment. Compared with the embodiment of FIG. 20, 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. 22 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. 3, the drive unit 36 is engaged with the base end portion of the dispensing tip 20 to hold the dispensing tip 20 and a syringe provided on the dispensing tip 20. The syringe drive part 36b which engages with the plunger 22 of 21 and drives a syringe is provided on the same axis, and both the movement of the dispensing tip 20 and the drive of the syringe 21 can be performed.

  FIG. 23 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 the reaction kit, (A) is a vertical sectional view, and (B) is a plan view showing a reaction plate and a dispensing tip. It is an external appearance perspective view of the same Example. 2 shows a dispensing tip 20 and a catch mechanism for mounting it, (A) is a state before attaching the catch mechanism to the dispensing tip, (B) is a state where the catch mechanism is attached to the dispensing tip, (C) has shown sectional drawing of the state which pulls up the plunger in a dispensing tip. In this embodiment, the catch mechanism 90 has a mounting operation (A1) to (A6) in which the dispensing tip 20 is mounted, and a vertical section showing the release operation (B1) to (B2) in which the catch mechanism 90 releases the dispensing tip 20. FIG. 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 Film 19 Dispensing nozzle 20 Dispensing tip 21 Syringe 22 Plunger 23 Filter 24 Cover 26 Cover body 28 Bellows film 32, 32a Sample container 36a Tip holder 36b Plunger holder 64, 64a, 71 Cover plate 66, 68, 72 Sealing material 90 Catch mechanism 92a, 94a Coil screw 93 Sleeve 95 Loop space 96 Isolation member 98a, 98b Packing 99 Air hole 100, 110, 120 DNA chip 106 Electrode 102 Electricity Flow path for electrophoresis separation

Claims (14)

A dispensing nozzle provided at the tip, connected to the upper part of the dispensing nozzle, and a syringe having a hollow inside, and a liquid suction / dispensing by the dispensing nozzle by sliding up and down in the syringe Equipped with a plunger for discharging,
A dispensing tip having an airtight property separating the inside of the nozzle from the outside and a flexibility allowing the plunger to slide between the upper portion of the plunger and the upper portion of the syringe.   The dispensing tip according to claim 1, wherein an air hole leading to a loop-shaped space formed by the syringe, the plunger, and the isolation member is provided. A reaction plate provided with a reaction vessel for causing the sample to react on the surface side;
The dispensing tip according to claim 1, which is disposed above the surface side of the reaction plate;
A cover that covers the plate upper space on the surface side of the reaction plate, and supports the dispensing tip so that the tip is located inside the plate upper space and the base end is located outside. ,
Reaction kit with   The dispensing tip is provided with an air hole that leads to a loop-shaped space formed by the syringe, the plunger, and the separating member, and the air hole is disposed inside the space above the plate covered by the cover. The reaction kit according to claim 3.   The reaction kit according to claim 3 or 4, further comprising a sample introduction part for injecting a sample into the space on the plate from the outside through an opening provided in a part of the cover so as to be hermetically sealed.   The reaction kit according to any one of claims 3 to 5, 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 3 to 6, wherein the dispensing tip includes a filter inside a tip portion.   The reaction kit according to any one of claims 3 to 7, wherein the reaction plate is provided with a gene amplification section for performing a gene amplification reaction on the surface side thereof.   The reaction kit according to any one of claims 3 to 8, wherein the reaction container is made of a light-transmitting material so that optical measurement can be performed from the bottom.   The reaction kit according to any one of claims 3 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 3 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 3 to 12, wherein the dispensing tip is held on the cover plate so as to be slidable in the vertical direction while being airtight with another sealing material.

Images