JP2018174908A - Reagent cassette - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a container for PCR or the like that can improve reliability and rapidity of PCR simply and stably.SOLUTION: A container for PCR 1 is mainly constituted of: a container body 11 which has a reaction chamber 12 for housing a specimen and reagent to cause rection in performing PCR on its bottom part 14; a lid body 6 which can be freely fitted to the container body 11; and a reagent cassette 16 housed in an area between an underside 7 of the lid body 6 and the reaction chamber 12 inside the container body 11. The reagent cassette 16 has at least one reagent housing part that can seal reagent by sealing means such as paraffin which is solid before PCR and melts by heating of PCR. With this configuration, when a specimen required for PCR is housed in the container body and PCR is performed in the state where reagent is previously housed in the reagent housing part and sealed, sealing means melts by heating of PCR, and the reagent housing part is opened. Then, the specimen and reagent move to the reaction chamber below.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a container for PCR, a container for PCR containing a reagent, and a reagent cassette, and more particularly to a container for PCR, a container for PCR containing a reagent, and a reagent cassette used when performing PCR with a DNA detection device.

  Conventionally, when amplifying DNA by polymerase chain reaction (herein referred to as "PCR"), a container for PCR as shown in FIG. 19 and FIG. 20 is used.

  FIG. 19 is a schematic front view showing the appearance of a conventional PCR container, in which (1) is in a state in which the lid is open, and (2) is in a state in which the lid is closed, FIG. 20 is a cross-sectional view showing a central cross-sectional structure of the PCR container shown in (2) of FIG. 19 with the lid closed.

  Referring to these drawings, PCR container 71 is made of, for example, polypropylene, and is mainly composed of container body 81, lid 76, and connection portion 77.

  The container body 81 has a cylindrical shape in which the upper side (the direction in which the lid is present with respect to the container body when the container for PCR is used) is opened, and the bottom is closed.

  The lid 76 is in the shape of a disk, and its size can completely cover the upper opening of the container body 81. The lid 76 has a circular ring-shaped locking portion 78 formed on its lower surface, which protrudes downward. As shown in FIG. 20, when the lid 76 is in a closed state, the lid 76 is open at the top of the container body 81 by the side wall outer surface of the locking portion 78 being locked with the inner wall surface of the container body 81. It is fixed in the state which covers completely. This prevents contamination of the container body 81 with foreign matter, and evaporation of the stored contents of the container body 81 during PCR reaching a maximum temperature of 90 ° C. or higher.

  The connection portion 77 is in the form of a flexible band within a predetermined angle range, and one end thereof is connected to the end of the lid 76 and the other end is connected to the end of the container main body 81. The lid 76 is fitted to the container main body 81 by bending the connection portion 77 and closing it.

  When using the PCR container 71, first, in a state in which the lid 76 as shown in (1) of FIG. In addition, necessary amounts of reagents (not shown) (for example, water, buffer, primers, dNTP, DNA polymerase and fluorescent reagent) necessary for the progress of PCR are introduced using a laboratory instrument such as a micropipette. Next, by pressing the upper surface of the lid 76 from above, the outer surface of the locking portion 78 of the lid 76 is locked with the inner surface of the container main body 81, and the lid 76 is shown in FIG. It is fixed in the closed state as shown. The PCR container 71 in this state is placed in a DNA amplification apparatus such as a thermal cycler (not shown), and the cycle of heating and cooling is repeated a predetermined number of times, whereby the PCR proceeds and the target DNA is amplified. After the PCR is completed, if necessary, the contents of the PCR container 71 containing the amplified target DNA can be collected in another container to detect the presence of the target DNA by light detection.

  Further, in Patent Document 1, the reagent used for the reaction is provided in the reaction container separately from the reaction region by the partition wall, and the partition wall is installed independently of the reaction solution so as to be temperature-controllable. Disclosed is a reaction container configured to be capable of releasing a reagent into a reaction region by melting the part by heating. The partition is made of wax, oil and the like and is disclosed as being horizontally installed in the reaction vessel.

JP, 2009-106221, A

  In the conventional container for PCR as described above, it is necessary to use an experimental instrument such as a micropipette in order to measure the amount of reagents necessary for the progress of PCR, which is complicated and requires the skill of the experimenter. There is a problem that the yield of PCR is greatly affected. In addition, there is also the possibility of contamination on the site or to the experimenter by reagents and PCR products. Therefore, a technique for obtaining stable measurement results easily and safely by PCR without a skilled person has been highly desired.

  Patent Document 1 suggested one of the means for solving the problem, but it was practically difficult to stably form the partition wall in the reaction vessel. That is, it is difficult to first form the partition in the reaction vessel using wax or the like in the molten state because the form is easily broken by the impact of dropping the wax in the molten state in the reaction vessel, and wax or the like in the solid state Even if it is intended to prepare a partition consisting of in advance and store it in the reaction container, it does not create any gap between the partition and the inner surface of the reaction container so that the reagent does not fall into the reaction area due to tilting etc. It was difficult to install the bulkhead.

  This invention was made in order to solve the above subjects, and it aims at providing the container etc. for PCR which can improve the certainty and rapidity of PCR simply and stably. .

  In order to achieve the above object, the invention according to claim 1 is a container for PCR, which has a reaction chamber for storing and reacting a sample and a reagent at the bottom of the container, and a container body And a reagent cassette housed at a position between the lower surface of the lid inside the container body and the reaction chamber, wherein the reagent cassette is solid prior to PCR and the heating of the PCR And at least one reagent storage portion capable of sealing the reagent by sealing means that melts.

  With this configuration, when a sample necessary for PCR is stored in the container body and PCR is performed in a state where the reagent is stored and sealed in advance in the reagent storage portion, the sealing means is melted by the heating of the PCR, and the reagent The storage unit is opened. Then, the sample and the reagent move to the lower reaction chamber.

  According to a second aspect of the present invention, in the configuration of the first aspect, the lower surface of the lid is formed with an upwardly recessed recess capable of storing at least a portion thereof by contact with the sample. The reagent storage portion is shaped to penetrate in the vertical direction.

  With this configuration, when the sample is stored using the concave portion on the lower surface of the lid and PCR is performed in a state where the reagent is stored and sealed in advance in the reagent storage portion, the sealing means is melted by the heating of the PCR. The reagent storage portion penetrates in the vertical direction. Then, the sample moves from the recess on the lower surface of the lid to the reagent storage unit, and the sample and the reagent move to the lower reaction chamber.

  According to a third aspect of the present invention, in the configuration of the first or second aspect of the present invention, in the reagent cassette, a plurality of reagent storage portions are provided, and the plurality of reagent storage portions are separated by a partition wall. .

  According to this structure, the plurality of reagent storage units can be sealed in a state in which the reagents stored in each of the plurality of reagent storage units are not mixed.

  According to the invention of claim 4, in the configuration of the invention according to any one of claims 1 to 3, the reagent cassette has a cylindrical shape opened in the vertical direction, and extends in the vertical direction to the side wall thereof. A slit communicating with the storage portion is formed.

  With this configuration, the surface area of the portion from which the reagent in the reagent cassette flows out is increased.

  According to the invention of claim 5, in the constitution of the invention according to any one of claims 1 to 4, the reaction chamber has a quadrangular frustum shape and is made of a transparent material.

  With this configuration, a surface facing the reaction chamber is generated.

  In the invention according to claim 6, in the configuration of the invention according to any one of claims 1 to 5, the container body, the lid and the reagent cassette are a lid when the lid is fitted to the container body. The dimensional relationship is set such that a gap is generated between the lower surface of the reagent cassette and the lower surface of the reagent cassette.

  With such a configuration, it is possible to seal the gap by the sealing means at the time of use.

  In the invention according to claim 7, in the constitution of the invention according to any one of claims 1 to 6, at least a part of the inner surface of the container main body protrudes inwardly toward the reagent cassette. .

  With this configuration, positioning of the reagent cassette inside the container body is facilitated.

  According to an eighth aspect of the present invention, in the configuration of the invention according to any one of the first to seventh aspects, the container main body has a thickness of 0.05 mm or more and 0.5 mm or less.

  With this configuration, the thickness of the portion of the container body where the reagent cassette is stored and the thickness of the reaction chamber are 0.05 mm or more and 0.5 mm or less.

  The invention according to claim 9 is the configuration of the invention according to any of claims 1 to 8, wherein the sealing means is wax, grease, paraffin or wax.

  When configured in this manner, the sealing means is solid at room temperature, and becomes a sealing means that becomes liquid at the first heating of PCR.

  According to the invention of claim 10, in the constitution of the invention according to any one of claims 1 to 9, the reagent cassette is formed with a concave portion concaved in at least one of its upper surface and lower surface. It is

  With this configuration, an inwardly concave portion is formed on the upper or lower surface of the reagent cassette.

  According to an eleventh aspect of the present invention, in the configuration of the invention according to any one of the first to tenth aspects, at least a portion of the lid can be accommodated on the lower surface of the lid by contact with the specimen. A concave recess is formed, the reagent storage portion is shaped to penetrate in the vertical direction, a convex convex portion is formed on the upper surface of the reagent cassette, and the convex portion is When the lid is fitted to the container body, it enters at least a part of the recess.

  With this configuration, the sample stored in the recess on the lower surface of the lid is easily scraped out during use.

  According to a twelfth aspect of the present invention, in the configuration of the invention according to any one of the first to eleventh aspects, a downwardly convex portion is formed on the lower surface of the reagent cassette.

  With this configuration, the lower surface of the reagent cassette is not horizontal but convex toward the reaction chamber.

  The invention according to claim 13 is a container for PCR containing a reagent using the PCR container according to any one of claims 1 to 12, wherein the reagent cassette contains the reagent in the reagent storage unit, It is sealed by the sealing means.

  With this configuration, the reagent cassette is stored and sealed in advance.

  The invention described in claim 14 is the configuration of the invention described in claim 13, wherein the reaction chamber is filled by a sealing means.

  With this configuration, the reaction chamber is in a state where air is discharged before PCR, and the sealing means melts in PCR and the sample and the reagent are stored via the reagent cassette.

  The invention according to claim 15 is a reagent cassette housed inside the container for PCR, comprising a reagent storage unit, wherein the reagent storage unit stores reagents necessary for PCR and is solid before PCR It is sealed by the sealing means which melts by the heating of PCR.

  With this configuration, the reagent cassette is stored and sealed in advance.

  The invention according to claim 16 is a container for PCR, which comprises a container body having a reaction chamber for storing and reacting a sample and a reagent at the bottom of the container, and a lid which can be fitted to the container body. The lid has at least one reagent storage formed at the tip through the exposed part of the lower surface of the tip, and the other part of the bottom of the tip is a specimen and The upper part is formed with the concave part which can store at least one part by the contact of the reagent, and the reagent storage part is a solid state before PCR, and the exposed part is a sealing means which is melted by the heating of PCR. Are sealable.

  With this configuration, when the sample is stored using the concave portion on the lower surface of the lid and PCR is performed in a state where the reagent is stored and sealed in advance in the reagent storage portion, the sealing means is melted by the heating of the PCR. , The reagent storage unit and the recess are opened. Then, the sample and the reagent move to the lower reaction chamber.

  According to a seventeenth aspect of the present invention, in the configuration of the sixteenth aspect of the present invention, an opening passing through the reagent storage portion is formed on the side wall of the tip portion.

  With this configuration, it is possible to inject the reagent into the reagent storage unit from the opening formed in the side wall of the tip end.

  As described above, according to the first aspect of the present invention, when the sample necessary for PCR is stored in the container body and PCR is performed in a state where the reagent is stored and sealed in advance in the reagent storage portion, heating of the PCR is performed. Thus, the sealing means is melted and the reagent storage portion is opened. And, since the sample and the reagent move to the lower reaction chamber, it becomes a container for PCR which can improve the certainty and rapidity of PCR simply and stably.

  According to the second aspect of the present invention, in addition to the effects of the first aspect, in a state where the reagent is stored and sealed in advance in the reagent storage portion, the sample is stored using the concave portion on the lower surface of the lid. When PCR is performed, the sealing means is melted by the heating of the PCR, and the reagent storage portion penetrates in the vertical direction. Then, since the sample moves from the recess on the lower surface of the lid to the reagent storage unit, and the sample and the reagent move to the lower reaction chamber, the convenience, reliability and quickness of PCR can be further improved. It becomes a container.

  In the invention according to claim 3, in addition to the effects of the invention according to claim 1 or claim 2, the plurality of reagent containing parts can be sealed in a state in which the reagents contained in each are not mixed. , It becomes easy to use a reagent which is not suitable for mixing in advance.

  According to the fourth aspect of the invention, in addition to the effect of the invention according to any one of the first to third aspects, the surface area of the portion from which the reagent flows out of the reagent cassette is increased, so from the reagent cassette to the reaction chamber It is easy for the reagent to drop.

  According to the fifth aspect of the invention, in addition to the effect of the invention according to any one of the first to fourth aspects, a surface opposite to the reaction chamber is generated, so in the horizontal direction relative to the reaction chamber after PCR The certainty of light detection when light detection is performed is improved.

  According to the sixth aspect of the invention, in addition to the effect of the invention according to any one of the first to fifth aspects, since the gap can be sealed by the sealing means during use, the lid is fitted. When the reagent cassette is compressed, the risk of leakage of the stored reagent is reduced as compared with the case where the reagent cassette is set to a size relationship in which no gap is generated between the lower surface of the lid and the reagent cassette.

  According to the seventh aspect of the invention, in addition to the effect of the invention according to any one of the first to sixth aspects, positioning of the reagent cassette inside the container body is facilitated, so that the reagent cassette can be moved to the reaction chamber It becomes possible to guide the reagent to a position where it is easy to drop, and the reliability of the PCR is improved.

  According to the eighth aspect of the invention, in addition to the effect of the invention according to any one of the first to seventh aspects, the thickness of the portion where the reagent cassette is accommodated in the container body and the thickness of the reaction chamber is 0.05 mm or more Since the size is less than .5 mm, the sealing means appropriately melts using PCR heating and PCR is properly performed.

  In addition to the effect of the invention according to any one of claims 1 to 8, the invention according to claim 9 is a sealing means which is solid at normal temperature and which becomes liquid upon initial heating of PCR. Therefore, the certainty of PCR is improved.

  According to the tenth aspect of the invention, in addition to the effects of the invention according to any one of the first to ninth aspects, since the concave portion is formed inward on the upper surface or the lower surface of the reagent cassette, the sealing is performed. The reliability of sealing by the stopping means is improved.

  According to the invention of claim 11, in addition to the effect of the invention according to any one of claims 1 to 10, the sample stored in the recess of the lower surface of the lid is easily scraped out during use. It becomes easy for the sample to reach the reaction chamber, and the reliability of the PCR is improved.

  In the invention according to claim 12, in addition to the effect of the invention according to any one of claims 1 to 11, since the lower surface of the reagent cassette is not horizontal but convex toward the reaction chamber, the reagent is It becomes easy to drop into the reaction chamber, and the reliability of the PCR is improved.

  In addition to the effect of the invention according to any one of claims 1 to 12, the invention according to claim 13 is a reagent cassette in which a reagent is stored and sealed in advance, so a desired amount and type of the reagent cassette can be obtained. These reagents can be stored in advance, and the reliability and speed of PCR can be improved simply and stably.

  According to the invention of claim 14, in addition to the effect of the invention of claim 13, the reaction chamber is in a state in which air is discharged before PCR, the sealing means is melted at the time of PCR, and the sample is inserted via the reagent cassette. And, since the reagent is stored, it becomes easy to introduce the sample and the reagent into the reaction chamber, and the reliability of the PCR is improved.

  Since the invention according to claim 15 is a reagent cassette in which reagents are stored and sealed in advance, a desired amount and type of reagent can be stored in advance, and it is simple and stable by using for a container for PCR In addition, the certainty and rapidity of PCR can be improved.

  The invention according to claim 16 is that sealing is performed by heating the PCR when the sample is stored using the concave portion on the lower surface of the lid while the reagent is stored in the reagent storage portion in advance and sealed. Is dissolved, and the reagent storage unit and the recess are opened. And, since the sample and the reagent move to the lower reaction chamber, it becomes a container for PCR which can improve the certainty and rapidity of PCR simply and stably.

  In addition to the effects of the invention of claim 16, the invention of claim 17 makes it possible to inject the reagent into the reagent storage unit from the opening formed in the side wall of the tip portion, so that the usability is improved. improves.

It is the front view which shows the external appearance shape and center cross-section of a container for PCR by the 1st embodiment of this invention, and a sectional view which fractured partially. It is the front view which shows the external shape and center cross-section of the container main body of the container for PCR shown in FIG. It is a side view which shows the external appearance shape of the container main body of the container for PCR shown in FIG. It is the front view which shows the external appearance shape and center cross-section of a lid of the container for PCR shown in FIG. 1, and a partially broken sectional view. It is a front view which shows the external appearance shape of the reagent cassette of the container for PCR shown in FIG. It is an end elevation of a VI-VI line shown in Drawing 5, and (1) is a state before storing a reagent, and (2) is a state after storing and sealing a reagent. It is a schematic front view which shows the state before operation | movement of the DNA detection apparatus using the container for PCR shown in FIG. It is a figure corresponding to FIG. 7, Comprising: It is a schematic front view which shows the operation state of a DNA detection apparatus. It is a schematic diagram of A part shown in FIG. It is a figure corresponding to FIG. 7, Comprising: It is a schematic front view which shows the state which PCR completed and the container for PCR moved to the detection part. It is an arrow view of the XI-XI line shown in FIG. It is the front view and the partially broken sectional view which show the external shape and center cross-section of the lower half part of the container for PCR by 2nd Embodiment of this invention. It is an end elevation of the XIII-XIII line shown in FIG. It is the front view which shows the external appearance shape and center cross-section of a lid of a container for PCR by a 3rd embodiment of this invention, and a partially cutaway sectional view. It is sectional drawing of the XV-XV line shown in FIG. 14, Comprising: (1) is a state before accommodating a reagent, (2) is a state after accommodating and sealing a reagent. FIG. 15 is an end view of the XVI-XVI line shown in FIG. It is a schematic plan view of a reagent cassette of a container for PCR according to another embodiment of the present invention. FIG. 16 is a schematic front view of a reagent cassette of a container for PCR according to still another embodiment of the present invention. It is a schematic front view which shows the external appearance shape of the conventional container for PCR, Comprising: (1) is a state of a lid open state, (2) is a state of a lid closed. FIG. 20 is a cross-sectional view showing a central cross-sectional structure of the PCR container shown in (2) of FIG. 19 with the lid closed.

  FIG. 1 is a front view and a partially broken cross sectional view showing the external shape and the central cross sectional structure of a PCR container according to a first embodiment of the present invention.

  Referring to the drawing, PCR container 1 is made of, for example, transparent polypropylene and has a circular ring-shaped horizontal cross section, and reaction chamber 12 in which a sample and a reagent are stored and reacted at the bottom portion 14 thereof is formed. Mainly from the container body 11 having the lid, the lid 6 which can be fitted to the container body 11, and the reagent cassette 16 housed at a position between the lower surface 7 of the lid 6 and the reaction chamber 12 inside the container body 11. It is configured. Further, in the figure, the male screw portion 13 formed on the upper outer side surface of the container main body 11 and the female screw portion 9 formed on the upper inner side surface of the lid 6 are in a sealed state in which they are screwed. It prevents the entry of foreign matter from the outside. By holding and rotating the knurled portion 8 at the upper part of the lid 6, the lid 6 can be shifted to the opened state in which the fitting between the lid 6 and the container main body 11 is released. It is possible to inject a sample into the inside of the container body 11, take out the reagent cassette 16 out of the container body 11, or store it in the container body 11.

  When the PCR container 1 is used, the reaction chamber 12 is positioned at the lowermost position in the vertical direction as shown in FIG. 1 to store and react the sample and the reagent. In the present specification, the upward direction refers to the direction in which the reagent cassette or lid is positioned relative to the reaction chamber when the PCR container is used, and the downward direction refers to the reagent cassette or lid when the PCR container is used. It means the direction in which the reaction chamber is located.

  And, in the container 1 for PCR, in the sealed state shown in FIG. 1, the reagent cassette 16 is placed on the flat portion 15 extending outward from the peripheral edge of the upper end of the reaction chamber 12 at the bottom portion 14 of the container body 11 At the same time, a part of the upper surface 17 of the reagent cassette 16 and a part of the lower surface 7 of the lid 6 abut without a gap. Thus, the reagent cassette 16 is fixed in position in the PCR container 1.

  Next, each component of the container 1 for PCR is demonstrated.

  FIG. 2 is a front view and a partially broken sectional view showing the external shape and central sectional structure of the container body of the PCR container shown in FIG. 1, and FIG. 3 is a container body of the PCR container shown in FIG. It is a side view which shows an external appearance shape.

  With reference to these figures, the container body 11 is in the form of a cylinder having an open top, and the reaction chamber 12 at the bottom 14 of the container body 11 is in the form of a transparent material which is in the form of a tapered square pyramid. It is done. By configuring in this manner, opposing surfaces are generated in the reaction chamber 12. The effect of this will be described later.

  FIG. 4 is a front view and a partially broken cross-sectional view showing an outer appearance shape and a central cross-sectional structure of a lid of the PCR container shown in FIG.

  With reference to the drawing, lid 6 is made of synthetic resin such as polypropylene, for example, and on the lower surface 7 thereof, an upwardly recessed concave portion 10 capable of storing at least a part thereof by contact with specimen 31 is formed. It is done. At the time of use, as shown by an arrow drawn by a dashed dotted line in the figure, a part (for example, a target) of the sample 31 is made in the recess 10 by bringing the recess 10 of the lid 6 into contact with the sample 31 (for example, intraoral mucous membrane) Saliva containing DNA) can be conveniently collected without using a laboratory instrument or the like. The effect of this will be described later.

  FIG. 5 is a front view showing the external shape of the reagent cassette of the PCR container shown in FIG. 1, and FIG. 6 is an end view of the VI-VI line shown in FIG. (2) is the state after storing and sealing the reagent.

  With reference to these figures, reagent cassette 16 is made of synthetic resin such as polypropylene, for example, and has a cylindrical shape penetrating in the vertical direction as a basic skeleton, and extends in the vertical direction on side wall 24 of reagent storage portions 21a to 21c. Each of the slits 23a-23c in communication with each of the is formed. The effect of this will be described later.

  The reagent cassette 16 also has reagent storage portions 21a to 21c which can seal the reagents 32a to 32c with paraffins 25a to 25c which are solid before PCR and melt by heating of the PCR. The melting point of paraffin is set at about 50.degree. That is, in the state where each of the reagent storage parts 21a to 21c stores each of the reagents 32a to 32c, the opening and the slit 23a to 23c (not shown) are sealed by sealing means such as paraffin 25a to 25c. Can. With this configuration, as shown in (2) of FIG. 6, the reagents 32a to 32c are stored in advance in the reagent storage portions 21a to 21c and sealed, and as shown in FIG. When PCR is performed by storing a sample necessary for PCR in the container body 11 in a sealed state in which the reagent cassette 16 is housed inside, the reagent cassette 16 itself made of synthetic resin is melted by the heating of PCR reaching up to over 90 ° C. The paraffins 25a to 25c, etc. are melted without any reaction, and the reagent storage portions 21a to 21c are opened downward. Then, the sample and the reagents 32 a to 32 c move to the lower reaction chamber 12. Therefore, the container for PCR of the present invention is simple because it does not use an experimental device such as a micropipette, and is stable because the reagent cassette has a basic skeleton (side wall) made of synthetic resin, which is an experiment. It is possible to eliminate the trouble of reagent injection by a person, prevent measurement error and contamination, and improve the reliability and quickness of PCR.

  When the reagent is stored, for example, the lower opening (not shown) of the reagent cassette 16 and the slits 23a to 23c are pressed against the melted paraffin and then cooled to lay down the paraffin, and a desired amount of the reagent 32a to 32c After the 32c is accommodated from above by an automatic dispenser not shown, the upper opening of the reagent cassette 16 is also sealed by filling with paraffin.

  Furthermore, as described above with reference to FIG. 4, the recess 10 is formed on the lower surface 7 of the lid 6, and the reagent storage portions 21 a to 21 c are vertically penetrated. When 25c melts, the reagent storage portions 21a to 21c penetrate in the vertical direction. Then, as described above with reference to FIG. 1, a part of the upper surface 17 of the reagent cassette 16 and a part of the lower surface 7 of the lid 6 abut each other without a gap. The reagent and reagents 32a to 32c move to the lower reaction chamber 12 via the reagent storage units 21a to 21c. Therefore, the simplicity, certainty and rapidity of PCR can be further improved.

  Further, in the reagent cassette 16, a plurality of reagent storage sections 21a to 21c are provided, and these are partitioned by the respective partitions 22a to 22c. The partition walls 22a to 22c are plate-like members extending in the vertical direction from the upper surface to the lower surface (not shown) of the reagent cassette 16, and are arranged to form an angle of 120 ° with each other in plan view. It is divided into volumes. As a result, as shown by (2) in the figure, sealing can be performed in a state in which the reagents 32a to 32c stored in each of the reagent storage portions 21a to 21c are not mixed. Therefore, it becomes easy to use a reagent which is not suitable for mixing in advance.

  The reagent is a reagent necessary for PCR, and may be selected from a desired type depending on the use, and examples thereof include water, buffer, primer, dNTP, magnesium compound, DNA polymerase, fluorescent reagent and the like. Although the fluorescent reagent is not necessary for the PCR itself, for the purpose of simultaneously performing light detection in the DNA detection apparatus after completion of the PCR, the embodiment of the present invention will be described including the fluorescent reagent.

  Furthermore, as described above, the reagent cassette 16 has a cylindrical shape opened in the vertical direction, and each of the slits 23a to 23c extending in the vertical direction on the side wall 24 and communicating with each of the reagent storage portions 21a to 21c is formed It is done. As a result, the surface area of the portion of the reagent cassette 16 from which the reagents 32a to 32c flow out is increased, and the reagents 32a to 32c easily drop from the reagent cassette 16 toward the reaction chamber 12.

  Next, a process of causing PCR to proceed with a DNA detection device using the PCR container 1 containing a sample and a reagent will be described.

  FIG. 7 is a schematic front view showing a state before operation of the DNA detection device using the PCR container shown in FIG.

  Referring to the figure, DNA detection apparatus 91 performs detection by light detection of DNA amplification structure 92 for advancing PCR by heating and cooling, and PCR products stored in the PCR container in which PCR is completed. It mainly comprises a unit 112 and a control unit (not shown).

  The DNA amplification structure 92 includes a container holder 97 for holding the PCR container 1, a support portion 98 for supporting the container holder 97, a plate-shaped first heater 101 on the high temperature side, and an upper portion of the first heater 101. Mainly composed of a ring plate-shaped heating plate 102 fixed to the plate, a plate-shaped second heater 106 on the low temperature side, and a pedestal 110 for fixing the first heater 101 and the second heater 106 on the upper surface thereof. It is done.

  The PCR container 1 houses the reagent cassette 16 containing the reagent as described above, and is sealed by the lid 6 in a state where the sample is housed by the concave portion 10 of the lid 6 or manually.

  The container holder 97 holds the PCR container 1 so that the bottom surface of the PCR container 1 (reaction chamber 12) is horizontal to the surfaces of the first heater 101 and the second heater 106.

  The support 98 is configured to support the container holder 97. Further, in accordance with the instruction from the control unit, the support portion 98 moves in the left-right direction (the direction from the first heater 101 to the second heater 106 or its direction as shown by the arrow drawn by the dashed line in the upper part of the figure). It is possible to slide the PCR container 1 above the first heater 101, above the second heater 106, or in the opposite direction, from these, toward the detection unit 112 or in the opposite direction). It can be moved freely to the part 112.

  The first heater 101 is made of, for example, a heating body covered with silicone rubber, and its temperature can be adjusted by the control unit, and a recess 104 in which the reaction chamber 12 of the PCR container 1 can be fitted is formed on the top surface thereof. Have. The surface of the recess 104 is set to a high temperature of, for example, 120 ° C. by the control unit.

  Further, the heating plate 102 is made of, for example, aluminum having high thermal conductivity, and has a through hole 103 in the center of which the PCR container 1 can be inserted. The thickness of the heating plate 102 is substantially the same as the height of the reagent cassette 16 in the vertical direction. The inner surface of the through hole 103 is warmed by heat conduction from the first heater 101 and reaches, for example, 80.degree.

  The second heater 106 is the same as the first heater 101 described above except for the temperature, and comprises, for example, a heating body covered with silicone rubber, and the temperature thereof is adjustable by the control unit. The upper surface has a recess 109 into which the reaction chamber 12 of the PCR container 1 can be inserted. The surface of the recess 109 is set to a low temperature of, for example, 50 ° C. by the control unit.

  The pedestal 110 has a first heater 101 and a second heater 106 fixed on the top surface thereof. In addition, the pedestal 110 slides in the vertical direction (the direction from the reaction chamber 12 to the recess 104 in the state before the operation in FIG. 7 or the opposite direction thereof) as indicated by the arrow drawn by the alternate long and short dash line in the lower part of the figure. It is freely controlled. Thereby, the first heater 101 and the second heater 106 can be raised or lowered.

  FIG. 8 is a view corresponding to FIG. 7 and is a schematic front view showing the operating state of the DNA detection apparatus.

  Referring to the drawing, from the state before the operation shown in FIG. 7, the pedestal 110 ascends in the direction of the arrow drawn by the solid line in the drawing according to the instruction from the control unit, whereby the PCR container 1 is heated plate While passing through the through hole 103 of 102, the reaction chamber 12 of the PCR container 1 is fitted into the recess 104 of the first heater 101.

  Then, the portion of the PCR container 1 in which the reagent cassette 16 is housed is warmed by the heating of the PCR through the through hole 103 to melt the paraffin above and below the reagent cassette 16, and the reaction in the lower part of the reagent or sample therein. It is discharged into the chamber 12 (see also FIG. 9 described later). Further, the specimen and the reagent stored in the reaction chamber 12 of the PCR container 1 by the contact (first contact) of the surface of the reaction chamber 12 of the PCR container 1 with the surface of the recess 104 of the first heater 101. Is heated.

  The state of heating of the sample and the reagent by the first contact will be described in detail.

  FIG. 9 is a schematic view of a portion A shown in FIG.

  Referring to the drawing, first, as described above, the reagent cassette 16 is heated by the through holes 103 and paraffin in the upper and lower surfaces (not shown) melts and penetrates in the vertical direction. Therefore, as shown by the arrow drawn by the solid line in the figure, the sample stored in the recess 10 moves to the reagent storage portion 21. Furthermore, the sample and the reagent stored in the reagent storage unit 21 move to the reaction chamber 12 and become the PCR solution 33 which is a sample and reagent necessary for PCR.

  And since the surface of the first heater 101 is made of silicone rubber, it is excellent in elasticity (shape recovery from compressive deformation) and thermal conductivity. Therefore, in the first contact, the reaction chamber 12 is inserted into the recess 104 with high adhesion, and the PCR solution 33 is heated by direct heat conduction from the first heater 101 to the reaction chamber 12 Be done.

  In addition to this, in the PCR solution 33 during heating, the temperature is relatively low because the upper part of the PCR solution 33 shown by L is outward, and the lower part of the PCR solution 33 shown by H is from the first heater 101 The temperature is relatively high because the heat of the above is strongly transmitted, and the middle portion of the PCR solution 33 shown by M has a temperature about halfway between L and H. As a result, as shown by the arrows in the figure, the heated lower solution starts to rise and causes convection that is folded back on the surface of the ceiling. The heat convection efficiently heats the PCR solution 33. By the first contact, the PCR solution 33 is heated, for example, to 92 ° C. to denature the DNA.

  When heating by the first contact for a predetermined time is completed, the pedestal 110 descends according to an instruction from the control unit, the first contact is released, and the state shown in FIG. 7 is returned to. Hereinafter, although illustration is omitted, the support unit 98 horizontally moves the container holder 97 so that the PCR container 1 is above the second heater 106 according to an instruction from the control unit.

  Then, as in the case of the first contact described above with reference to FIG. 8, the reaction chamber 12 of the PCR container 1 is fitted into the recess 109 of the second heater 106 by raising the pedestal 110, and the PCR container 1 is obtained. A contact (second contact) with the surface of the reaction chamber 12 and the surface of the recess 109 of the second heater 106 occurs. By this second contact, the PCR solution 33 is cooled to, for example, 65 ° C., and annealing is performed.

  When the cooling by the second contact for a predetermined time is completed, the pedestal 110 descends according to the instruction from the control unit, and the container holder 97 is horizontally moved by the support unit 98, and the PCR container 1 is above the first heater 101. It returns to the state shown in FIG. 7 located.

  By alternately repeating the first contact and the second contact in this manner, PCR proceeds. Then, when the first contact and the second contact are repeated a predetermined number of times and PCR is completed, the process proceeds to the step of detecting amplified DNA.

  FIG. 10 is a diagram corresponding to FIG. 7 and is a schematic front view showing a state in which the PCR is completed and the PCR container is moved to the detection unit.

  Referring to the figure, after PCR is completed, support vessel 98 moves container holder 97 in the direction shown by the arrow in the figure according to the instruction from the control unit, and container 1 for PCR is moved to detection unit 112. .

  FIG. 11 is an arrow view of the XI-XI line shown in FIG.

  Referring to the drawing, detection unit 112 mainly includes light emitting element 113 and light receiving element 114. The light emitting element 113 and the light receiving element 114 are arranged to face each other on the same straight line in the horizontal direction. The PCR container 1 is positioned so that the straight line penetrates the reaction chamber 12 in which the amplified DNA is stored. The light emitting element 113 is an LED lamp having an effective wavelength of 450 nm to 570 nm. The light receiving element 114 is a photodiode having an effective wavelength of 300 nm to 820 nm.

  At the time of detection, the light emitting element 113 emits excitation light toward the reaction chamber 12 of the PCR container 1 according to an instruction from the control unit (not shown). The amplified DNA in the reaction chamber 12 is bound to a fluorescent reagent contained in the reagent, and reacts with excitation light to fluoresce. Since the reaction chamber 12 is made of a transparent material, it can transmit light and receive the light emission intensity of the DNA from which the light receiving element 114 is fluorescent. This enables light detection of DNA without transferring the PCR product from the PCR container to another detection container. Then, based on the received light emission intensity, quantitative measurement of amplified DNA can be performed.

  Here, as described above, the reaction chamber 12 of the PCR container 1 has a quadrangular frustum shape and is made of a transparent material, so that a surface facing the reaction chamber is generated. The reliability of light detection when light detection is performed in the horizontal direction with respect to the room is improved.

  As described above, by providing the reagent cassette 16, the PCR container 1 can easily and stably improve the reliability and the rapidity of the PCR, and the concave portion of the lower surface 7 of the lid 6. The synergetic effect with 10 makes it possible to obtain stable measurement results easily and safely, even by non-experts.

  Further, such a reagent cassette 16 can be stored in the inside of the PCR container 1 in a state where the reagent is stored in advance in the reagent storage portion 21 and sealed by sealing means such as paraffin. With such a configuration, since the reagent cassette is stored in advance and the reagent is sealed, a desired amount and kind of reagent can be stored in advance, and it is simple and stable by using the container for PCR. In addition, the certainty and rapidity of PCR can be improved.

  Next, FIG. 12 is a front view and a partially broken sectional view showing the external shape and the central sectional structure of the lower half of the PCR container according to the second embodiment of the present invention, and FIG. FIG. 7 is an end view of the indicated XIII-XIII line.

  The basic configuration of the PCR container 41 is the same as that of the PCR container 1 according to the first embodiment, so differences will be mainly described.

  With reference to these figures, container body 42, lid 43 and reagent cassette 44, when lid 43 is fitted to container body 42 as shown in FIG. It is set in such a dimensional relationship that a gap is generated between it and 44. With this configuration, the gap can be sealed by the sealing means at the time of use, so that when the lid 43 is fitted and the reagent cassette 44 is compressed in the vertical direction, Compared to the case where the gap is not generated between the lower surface 45 and the reagent cassette 44 (in the first embodiment described above), the risk of leakage of the stored reagent is reduced.

  Also, in the container body 42, a rib 46 which is a part of the inner surface thereof protrudes inward toward the reagent cassette 44. With this configuration, the positioning of the reagent cassette 44 in the container main body 42 becomes easy, and it becomes possible to guide the reagent from the reagent cassette 44 to the central position where the reagent tends to fall relative to the reaction chamber 47. Certainty is improved.

  Further, the inner surface of the bottom portion 48 of the container main body 42 is inclined downward toward the reaction chamber 47 from the portion on the inner side of the rib 46. This makes it easy for the reagent to drop from the reagent cassette 44 positioned by the rib 46 to the reaction chamber 47, thereby further improving the reliability of PCR.

  Next, FIG. 14 is a front view and a partially broken sectional view showing an appearance shape and a central sectional structure of a lid of a container for PCR according to a third embodiment of the present invention, and FIG. (1) is a state before storing a reagent, (2) is a state after storing and sealing a reagent, and FIG. 16 is shown in FIG. FIG. 7 is an end view of the XVI-XVI line.

  The basic configuration of the PCR container according to the third embodiment is the same as that of the PCR container 1 according to the first embodiment, so differences will be mainly described.

  With reference to these drawings, lid 53 has a shape in which a reagent cassette is integrated at its tip 54.

  That is, the container for PCR according to the third embodiment (not shown) can be fitted to the container main body (not shown) having a reaction chamber for storing and reacting the sample and the reagent at the bottom of PCR. The lid 53 has reagent storage parts 57a to 57c formed in the tip 54 through the exposed part 56 of a part of the lower surface 55 of the tip 54. On the other part of the lower surface 55, an upwardly recessed recess 58 is formed which can receive at least a part by contact with the sample, and the reagent storage parts 57a to 57c are solid before PCR The exposed portion 56 can be sealed by a sealing means that is shaped like a mold and melted by the heating of PCR.

  With this configuration, when the reagent is stored and sealed in advance in the reagent storage portions 57a to 57c, PCR is performed by storing the sample using the concave portion 58 of the lower surface 55 of the lid 53. The sealing means is melted by heating, and the reagent storage portions 57a to 57c and the recess 58 are opened downward. Then, the sample and the reagent move to the lower reaction chamber (not shown). Therefore, it has become a container for PCR that can simply and stably improve the certainty and rapidity of PCR.

  Further, an opening 60 penetrating the reagent storage portion 57 is formed in the side wall 59 of the tip portion 54. This makes it possible to inject the reagent into the reagent storage portion 57 from the opening 60 formed in the side wall 59 of the distal end portion 54, thereby improving the usability.

  That is, when using the container for PCR according to the third embodiment, the lower surface 55 of the tip portion 54 of the lid 53 is first sealed with paraffin 61 in a molten state, and the exposed portion 56 is sealed. At this time, even if paraffin 61 enters into the recess 58, there is no problem as long as a portion in a state of being recessed upward remains. Then, after the reagent is injected from the opening 60 into the reagent storage unit 57, the opening 60 is sealed with paraffin 61, whereby the reagent is sealed as shown in (2) of FIG.

  Next, FIG. 17 is a schematic plan view of a reagent cassette of a container for PCR according to another embodiment of the present invention.

  Referring to the drawing, reagent cassette 63 shown in (1) has the same shape as the tip portion according to the third embodiment. The reagent storage part is divided into three parts with a volume ratio of 1: 1: 2 by the partition wall. At the time of use, it is possible to appropriately allocate the storage amount of the reagent as compared with the first embodiment.

  In the reagent cassette 64 shown in (2), the reagent storage part is divided into four parts at a volume ratio of 1: 1: 1: 1 by the partition wall.

  In the reagent cassette 65 shown in (3), the reagent storage part is divided into five parts by four partitions which do not pass through the center and form a planar view rhombus.

  In the reagent cassette 66 shown in (4), the reagent storage portion is divided into four by three partitions forming three parallel equidistant lines.

  In the reagent cassette 67 shown in (5), the reagent storage portion is divided into a plurality of nets by a plurality of partition walls.

  In the reagent cassette 68 shown in (6), a plurality of reagent storage portions penetrate in the vertical direction in the form of a vascular bundle with respect to the reagent cassette 68 in which the portion excluding the reagent storage portion is solid. In this case, the solid part of the reagent cassette 68 is a partition.

  The aspects of (1) to (6) described above are applicable to any of the reagent cassettes of the first to third embodiments (the tip of the lid in the case of the third embodiment). It is also possible to change the angle between the partitions, change the vertical angle of each partition, or combine the divisions of these partitions with each other. Furthermore, a slit may be formed on the side wall of each reagent cassette.

  Next, FIG. 18 is a schematic front view of a reagent cassette of a container for PCR according to still another embodiment of the present invention.

  Referring to the drawing, the reagent cassette 123 of (1) is formed with a concave portion 127 which is recessed inward (downward) on the upper surface 129 thereof. Similarly, a concave portion may be formed in the lower surface inward (upper), or a concave portion may be formed on the upper surface 130 and the lower surface 131 as in the reagent cassette 124 shown in (2). Also good. With this configuration, a concave portion is formed inwardly on the upper surface or the lower surface of the reagent cassette, so that sealing means such as paraffin can easily be retained in the concave portion when sealing, and sealing is performed. The reliability of sealing by means is improved.

  The reagent cassette 124 of (2) is an aspect in which all the circumferential ribs 132a and 132b which protrude outward are formed on the periphery of each of the upper surface 130 and the lower surface 131.

  On the lower surface 133 of the reagent cassette 125 of (3), a convex portion 134 having a convex shape is formed downward. With this configuration, the lower surface of the reagent cassette is not horizontal but convex toward the reaction chamber, so that the reagent is more likely to fall into the reaction chamber than in the case where the lower surface is horizontal, which ensures the PCR Improves the quality.

  A convex convex portion 136 is formed on the upper surface 135 of the reagent cassette 126 of (4). Here, for example, in the PCR container 1 according to the first embodiment, the recess 10 is formed on the lower surface 7 of the lid 6 as described above, and the reagent storage portion 21 penetrates in the vertical direction. It is a shape. The convex portion 136 is set so as to enter at least a part of the recess 10 when the lid 6 is fitted to the container body 11. With this configuration, the sample stored in the recess on the lower surface of the lid is easily scraped out during use, so that the sample can easily reach the reaction chamber, and the reliability of the PCR is improved.

  Even if a convex convex portion is formed outward (in the upper surface of the reagent cassette but in the lower surface of the reagent cassette) on both the upper surface and the lower surface of the reagent cassette. good. In addition, the case where the reagent cassette has a spherical shape is also included in this.

  In each of the above embodiments, the reagent cassette has a cylindrical shape, but may have another shape. For example, it may be a cylindrical shape having a cross-sectional polygonal ring shape or may have a spherical shape. The reagent cassette preferably has a cylindrical shape from the viewpoint of ensuring the orientation when stored in the container main body, but even in the case of, for example, a spherical shape, a large number of penetrations in multiple directions such as up, down, left, and right By providing a hole, it can implement suitably, making orientation determination unnecessary.

  In each of the above-described embodiments, the container for PCR and the container body, the lid, and the reagent cassette constituting the same have a specific shape, but may have other shapes. For example, the capacity may be changed according to the application.

  Furthermore, in each of the above-described embodiments, the reagent cassette has a plurality of reagent storage units, and the plurality of reagent storage units are separated from one another by the partition walls, but the reagent cassette includes at least one reagent storage unit. You should have it.

  Furthermore, in the above embodiments, the recess is formed on the lower surface of the lid, but the recess may not be formed. As long as the reagent cassette is provided, the present invention is applicable even when manually loading the sample.

  Furthermore, in each of the above-described embodiments, the reagent storage portion penetrates in the vertical direction, but may be exposed in at least one direction.

  Furthermore, in each of the above embodiments, the slits are formed on the side walls of the reagent cassette so as to extend in the vertical direction and divide the side walls, but the slits are formed to communicate at least with the reagent storage unit It should be good. Moreover, the slit may not be formed.

  Furthermore, in each of the above embodiments, the reaction chamber has a quadrangular frustum shape and is made of a transparent material, but the reaction chamber has another shape such as a hemispherical shape convex downward. The present invention can be applied without affecting the PCR itself even if it is not transparent material.

  Furthermore, in the second embodiment described above, the four ribs are formed on the inner surface of the container main body so as to protrude inward toward the reagent cassette, but the number and the positional relationship of the ribs may be appropriately determined. It may be changed, and at least a part of the inner surface of the container body may be protruded inward.

  Furthermore, in each of the above embodiments, the thickness of the container body is preferably 0.05 mm or more and 0.5 mm or less. With this configuration, the thickness of the reaction chamber and the portion where the reagent cassette is stored in the container main body is 0.05 mm or more and 0.5 mm or less, so the sealing means appropriately melts using PCR heating. In addition, PCR is properly performed.

  Furthermore, in each of the above embodiments, paraffin was used as the sealing means, but the sealing means may be wax, grease, paraffin or wax. These are all solid at normal temperature, and serve as a sealing means that becomes liquid at the time of the first heating of PCR, so that the reliability of PCR is improved. In addition, with solid form, semi-solid, glass-like, etc. are included if it can be sealed. Further, the liquid state (melted state) may be a state in which the fluidity is higher than the solid state and the sealing is released.

  Furthermore, in each of the above embodiments, the melting point of the sealing means is about 50 ° C., but the melting point (the temperature for transition to the above-described molten state) is preferably 40 ° C. to 80 ° C. More preferably, the temperature is 45 ° C to 60 ° C. Moreover, it is preferable that specific gravity is a thing smaller than water. By this, it is solid at normal temperature, and the reagent or sample can be isolated, melted by the first heating of PCR, and the function and effect of guiding the reagent or sample to the lower reaction chamber can be exhibited more reliably.

  Furthermore, in each of the above embodiments, although the reagent is not stored in the reagent storage portion of the reagent cassette, for example, the reagent is stored at the time of sale and is sealed by the sealing means. It may be. With this configuration, since the reagent cassette is stored and sealed in advance, the desired amount and type of the reagent can be stored in advance, and the reliability and speed of the PCR can be easily and stably. It is possible to improve the quality.

  Furthermore, in each of the above-described embodiments, the reaction chamber was not stored until starting the PCR, or only the sample was stored. However, degassing was caused by chemical or physical processing. The pressure may be reduced or vacuumed.

  Further, the reaction chamber may be filled by the sealing means. With this configuration, the reaction chamber is in a state in which air is discharged before PCR, and the sealing means melts during PCR and the sample and the reagent are stored via the reagent cassette. This makes it easier to guide the PCR and improve the certainty of the PCR.

  Furthermore, in the third embodiment described above, although the opening is formed in the side wall of the tip portion, the present invention is applicable even if the opening is not formed.

  Furthermore, in each of the above embodiments, although the reagent is stored in all the reagent storage sections of the reagent cassette, there may be a reagent storage section in which no reagent is stored at the time of use.

  Furthermore, in each of the above embodiments, the container for PCR was made of polypropylene, but from polyethylene, polystyrene, polyethylene terephthalate, polyethylene naphthalate, polycarbonate, polyamide, acrylic, cyclic polyolefin and aromatic nylon It is preferable that it consists of at least 1 sort (s) of raw material chosen from the group which consists of. This is because the entire PCR container, including the reagent cassette, is not deformed by heating of the PCR.

  Furthermore, in each of the above embodiments, a specific DNA detection device is used for amplification of DNA and detection of amplified DNA, but other DNA amplification devices or devices for detecting amplified DNA may be used. It is good.

DESCRIPTION OF SYMBOLS 1, 41 ... Container for PCR 6, 43, 53 ... Lid 7, 45, 55 ... Lower surface 10, 58 ... Concave part 11, 42 ... Container main body 12, 47 ... Reaction chamber 14, 48 ... Bottom part 16, 44, 63- 68, 123 to 126 Reagent cassette 17, 129, 130, 135 Upper surface 21, 57 Reagent storage 22 22 Partition 23 Slit 24 Side wall 25 61 Paraffin 31 Sample 32 Reagent 46 Rib 54 End portion 56: Exposed portion 59: Side wall 60: Opening 127, 128: Concave portion 131, 133: Lower surface 132: All circumferential rib 134, 136: Convex portion In the figures, the same reference numerals indicate the same or corresponding portions .

This invention relates to a reagent cassette, in particular, to a reagent cassette Ru use in performing PCR by DNA detection device.

The present invention was made in order to solve the problems as described above, and it is an object of the present invention to provide a reagent cassette that can improve the certainty and rapidity of PCR easily and stably.

In order to achieve the above object, the invention according to claim 1 is a reagent cassette housed inside a container for PCR, which comprises a basic skeleton and a basic skeleton and at least a part of which is exposed to the outside. And a reagent storage unit capable of storing reagents necessary for PCR .

With this configuration, the reagent is stored in the reagent storage unit, and the sample necessary for PCR is stored in the container in a state of being sealed by a sealing unit that is solid before PCR and is melted by heating the PCR. When PCR is performed, the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened. Then, the sample and the reagent move to the reaction chamber below the container.

According to the invention of claim 2, in the configuration of the invention of claim 1, the reagent storage portion is shaped so as to be exposed on the opposite surface .

According to this configuration, when the sample is stored on one side of the exposed portion of the reagent cassette, the sealing means is melted by the heating of the PCR, and the sample moves to the other side of the exposed portion with the opened reagent storage portion. Mix with reagents on the way.

According to a third aspect of the invention, in the structure of the invention of claim 1 or claim 2, wherein reagent accommodating portion is provided multiple pieces, the plurality pieces of the reagent storage section, the basic structure partitioned by partition walls from each other It is formed .

According to this structure, the plurality of reagent storage units can be sealed in a state in which the reagents stored in each of the plurality of reagent storage units are not mixed.
According to a fourth aspect of the present invention, in the configuration of the third aspect, three reagent storage portions are provided, and the partition walls are three plate members extending in the same direction, and adjacent ones of the plate members are provided. Each crosses to form an angle of 120 ° with each other, and the reagent storage parts are shaped to be exposed at both ends in the same direction.
When configured in this manner, three reagent storage sections each having the same volume are formed.

According to the invention of claim 5, in the configuration of the invention according to any one of claims 1 to 4 , the basic skeleton has a tubular shape in which both ends in the axial direction are open, and the side wall thereof has an axis A slit extending in the direction and in communication with the reagent storage portion is formed.

According to the invention of claim 6, in the constitution of the invention according to any one of claims 1 to 5 , at least a part of the lower surface of the basic skeleton is a PCR in the state of being housed inside the container for PCR. While being in contact with the inner surface of the container, at least a portion of the upper surface of the container is set in a dimensional relationship in contact with the lower surface of the lid of the PCR container .

With this configuration, the reagent cassette is restricted from moving up and down carelessly inside the PCR container.

Invention of claim 7, wherein, in the configuration of the invention according to any one of claims 1 to 6, the basic skeleton, at least one of the at least a portion, a concave portion recessed inwardly of the surface of the opposing Are formed.

With this configuration , the sealing means can easily stay in the concave portion when sealing.

The invention of claim 8, wherein, in the configuration of the invention according to any one of claims 1 to 7, reagent storage section, in a state housed inside PCR vessel, a shape that is exposed in a vertical direction The basic skeleton has a convex convex portion formed on at least a part of the upper surface thereof, and the convex portion is a lid of the container for PCR in a state of being housed inside the container for PCR It enters into at least a part of a recess formed on the lower surface of the body .

With such a configuration, the one stored in the concave portion of the lid of the PCR container is easily scraped out by the reagent cassette.

According to the invention of claim 9, in the constitution of the invention according to any one of claims 1 to 8 , the basic skeleton has an outwardly convex portion formed on at least a part of its one surface. It is

According to this structure, it is possible to project toward the reaction chamber rather than horizontally the lower surface of the reagent cassette.

The invention according to claim 10 is a reagent cassette housed in the inside of a container for PCR, which is formed of a basic skeleton and a basic skeleton, and is exposed to the outside through an exposed portion, and the reagent necessary for PCR is It comprises a reagent storage unit housed, and a sealing means which is solid before exposure to PCR and is sealed by melting by heating of PCR .

According to this structure, advance reagent Ri is Do a sealed reagent cassette while being accommodated, when accommodating a specimen required for PCR in vessel performing PCR, melting sealing means by heating PCR, reagents The storage unit is opened. Then, the sample and the reagent move to the reaction chamber below the container.

The invention according to claim 11 relates to the constitution of the invention according to claim 10, wherein the sealing means is wax, grease, paraffin or wax .

When configured in this manner, the sealing means is solid at room temperature, and becomes a sealing means that becomes liquid at the first heating of PCR.

According to the invention of claim 12, in the constitution of the invention of claim 10 or claim 11 , the basic skeleton is not melted by heating of PCR .

According to this structure, only the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened.

As described above, according to the invention of claim 1 , the PCR is carried out in the container in a state in which the reagent is stored in the reagent storage portion and sealed by a sealing means which is solid before PCR and is melted by heating of PCR. When the PCR necessary to store the necessary samples is performed, the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened. Then, since the sample and the reagent move to the reaction chamber below the container, it is possible to simply and stably improve the reliability and the rapidity of the PCR.

In the invention according to claim 2, in addition to the effect of the invention according to claim 1, when the sample is stored on one side of the exposed part of the reagent cassette, the sealing means is melted and opened by the heating of the PCR. Since the reagent is mixed with the reagent while the sample moves to the other side of the exposed portion in the reagent storage unit, it is possible to simply and stably improve the reliability and quickness of the PCR.

In the invention according to claim 3, in addition to the effects of the invention according to claim 1 or claim 2, the plurality of reagent containing parts can be sealed in a state in which the reagents contained in each are not mixed. , It becomes easy to use a reagent which is not suitable for mixing in advance.
According to the fourth aspect of the invention, in addition to the effects of the third aspect of the invention, three reagent storage portions each having the same volume are formed, so that the reagents are easily dropped.

The invention of claim 5, wherein, in addition to the effect of the invention according to any one of claims 1 to 4, the surface area of the portion where the reagent of the reagent cassette flows out is increased, easily fallen reagents.

In the invention according to claim 6 , in addition to the effect of the invention according to any one of claims 1 to 5 , the reagent cassette is restricted from inadvertently moving in the vertical direction inside the PCR container. , The stability of the reaction is improved.

According to the seventh aspect of the invention, in addition to the effect of the invention according to any one of the first to sixth aspects , the sealing means is easily retained in the concave portion when sealing, so that the reliability of sealing is ensured. Improve.

According to the eighth aspect of the invention, in addition to the effect of the invention according to any one of the first to seventh aspects, the one stored in the concave portion of the lid of the PCR container is easily scraped out by the reagent cassette. Therefore, the certainty of PCR is improved.

In the invention according to claim 9 , in addition to the effect of the invention according to any one of claims 1 to 8 , the lower surface of the reagent cassette can be convex toward the reaction chamber instead of being horizontal. The reagent easily falls into the reaction chamber, and the reliability of the PCR is improved.

The invention of claim 10 wherein the pre-reagent Ri is Do a sealed reagent cassette while being accommodated, when accommodating a specimen required for PCR in vessel performing PCR, melting sealing means by heating the PCR , The reagent storage unit is opened. And, since the sample and the reagent move to the reaction chamber below the container , a desired amount and type of reagent can be stored in advance, and by using the container for PCR, the reliability and quickness of the PCR can be simply and stably. It is possible to improve the quality.

In addition to the effects of the invention according to claim 10 , the invention according to claim 11 is a sealing means which is solid at normal temperature and becomes liquid when the PCR is initially heated, so that the reliability of the PCR is improved. Do.

In the invention according to claim 12 , in addition to the effect of the invention according to claim 10 or 11 , only the sealing means is melted by heating of PCR and the reagent storage portion is opened, so that the stability of the reaction is improved. Do.

According to this structure, only the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened.
The invention as set forth in claim 13 is the configuration of the invention as set forth in any of claims 1 to 9, wherein the basic skeleton has a cylindrical shape in which both ends in the axial direction are open, and the inside is in the axial direction It is partitioned by the extending partition wall, and at least one of both axial end edges of the partition wall is located more inward than at least one of corresponding open end surfaces of the basic skeleton.
According to this structure, the sealing means is likely to stay at a place from the axial edge of the partition wall to the open end face of the basic skeleton when sealing.

In the invention according to claim 12, in addition to the effect of the invention according to claim 10 or 11, only the sealing means is melted by heating of the PCR and the reagent storage portion is opened, so that the stability of the reaction is improved. Do.
According to the invention of claim 13, in addition to the effect of the invention according to any one of claims 1 to 9, at the point from the axial edge of the partition wall to the open end face of the basic skeleton when sealing Since the sealing means is easily retained, the reliability of the sealing is improved.

In order to achieve the above object, the invention according to claim 1 is a reagent cassette housed inside a container for PCR, comprising: a basic skeleton having a cylindrical shape in which both ends in the axial direction are open ; It is provided with a reagent storage unit which is formed and exposed to the outside through the exposed portion and stores reagents necessary for PCR, and the exposed portion is solid prior to PCR and sealed by melting by heating of PCR. means can be sealed by, the reagent storage section, provided with a plurality, the plurality of reagent retract and part includes a partition wall which are partitioned from each other, the basic skeleton, and the exposed portion to be sealed by a sealing means is formed by the basic skeleton and the partition are those that do not melt by heating the PCR.

With this configuration, the reagent is stored in the reagent storage unit, and the sample necessary for PCR is stored in the container in a state of being sealed by a sealing unit that is solid before PCR and is melted by heating the PCR. When PCR is performed, the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened. Then, the sample and the reagent move to the reaction chamber below the container. Further, a plurality of the reagent storage section, thereby enabling the sealing in that state without mixing housed reagents together in each. Further, only the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened.

According to the invention of claim 2, in the constitution of the invention of claim 1, the reagent storage portion has a shape exposed at both open end faces of the basic skeleton .

請 Motomeko 3 the described invention, in the structure of the invention of claim 1 or claim 2, wherein the reagent storage unit, three is provided, the partition wall is a 3 plate-like member extending in the same direction, adjacent The plate-like members cross each other so as to form an angle of 120 ° with each other, and the reagent storage portions are shaped so as to be exposed at both ends in the same direction.
When configured in this manner, three reagent storage sections each having the same volume are formed.

Those invention of claim 4, in the constitution of the invention described in any one of claims 1 to 3, in the side wall of the basic bone price, a slit communicating with the reagent storage section extending in the axial direction is formed It is.

According to the invention of claim 5, in the constitution of the invention according to any one of claims 1 to 4 , at least a part of the lower surface of the basic skeleton is a PCR in a state of being housed inside a container for PCR. While being in contact with the inner surface of the container, at least a portion of the upper surface of the container is set in a dimensional relationship in contact with the lower surface of the lid of the PCR container.

According to the invention of claim 6, in the constitution of the invention according to any one of claims 1 to 5 , the basic skeleton is formed with a concave portion which is inwardly recessed in at least one of its upper surface and lower surface. It is

The invention of claim 7, wherein, in the configuration of the invention according to any one of claims 1 to 6, the reagent storage section, in a state housed inside PCR vessel, be shaped to expose in the vertical direction The basic skeleton has a convex convex portion formed on at least a part of the upper surface thereof, and the convex portion is a lid of the container for PCR in a state of being housed inside the container for PCR And at least a part of the recess formed on the lower surface of

According to an eighth aspect of the invention, in the configuration of the invention according to any one of the first to seventh aspects , a downwardly convex portion is formed on the lower surface of the basic skeleton.

The invention according to claim 9 is a reagent cassette housed inside the PCR container, which is formed of a basic skeleton having a cylindrical shape whose both ends in the axial direction are open, a basic skeleton, and an exposed portion. The reagent storage unit in which the reagents necessary for PCR are stored, and a sealing means that is solid before exposure to PCR and is sealed by melting by heating of PCR ; housing section, provided with a plurality, the plurality of reagent storage section, a partition wall which are partitioned from each other, the basic skeleton is formed by the exposed portion sealed by the sealing means, the basic skeleton and septal wall, It does not melt by heating of PCR .

With this configuration, the reagent cassette is stored and sealed in advance, and when the sample necessary for PCR is stored in the container and PCR is performed, the sealing means is melted by the heating of the PCR, and the reagent storage unit Is released. Then, the sample and the reagent move to the reaction chamber below the container. Further, a plurality of the reagent storage section, the sealing available-with no mixing is housed reagents together in each. Further, only the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened.

The invention according to claim 10 is the configuration of the invention according to any of claims 1 to 9, wherein the sealing means is wax, grease, paraffin or wax.

PetitionRequest11The described invention claims claims 1 to10In the configuration of the invention described in any of the above, the basic skeleton isUp and downHas an open cylindrical shape with both endsUp and downPartitioned by a partition extending in theUpdirectionThe edge of the, Of the basic skeletonUpward openingFree end FaceRimoDownwardIt is located in
  When configured in this manner, the barrierUpOf the basic skeleton from the edge of the directionUpwardThe sealing means can easily stay at the place up to the open end face.

As described above, according to the invention of claim 1, the PCR is carried out in the container in a state in which the reagent is stored in the reagent storage portion and sealed by a sealing means which is solid before PCR and is melted by heating of PCR. When the PCR necessary to store the necessary samples is performed, the sealing means is melted by the heating of the PCR, and the reagent storage portion is opened. Then, since the sample and the reagent move to the reaction chamber below the container, it is possible to simply and stably improve the reliability and the rapidity of the PCR. Further, a plurality of the reagent accommodating portion, since the reagent with each other housed in each of which are to be sealable in a state not mixed, it is easy to use reagents unsuitable for that you mixing Me beforehand. Furthermore, since only Rifutome means by the heating of the PCR is opened reagent accommodating portion melts, thereby improving the stability of the reaction.

Invention 請 Motomeko 3 wherein, in addition to claim 1 or claim 2, according to the present description, each for three reagent storage portion of the same volume are formed, easily fall reagent.

According to the fourth aspect of the invention, in addition to the effect of the invention according to any one of the first to third aspects, the surface area of the portion from which the reagent flows out of the reagent cassette is increased, so that the reagent tends to fall off.

According to the fifth aspect of the invention, in addition to the effect of the invention according to any one of the first to fourth aspects, the reagent cassette is restricted from inadvertently moving in the vertical direction inside the PCR container. , The stability of the reaction is improved.

According to the sixth aspect of the invention, in addition to the effect of the invention according to any one of the first to fifth aspects, since the sealing means is easily retained in the concave portion when sealing, the reliability of sealing is ensured. Improve.

According to the seventh aspect of the invention, in addition to the effect of the invention according to any one of the first to sixth aspects, the substance accommodated in the concave portion of the lid of the container for PCR is easily scraped out by the reagent cassette. Therefore, the certainty of PCR is improved.

In the invention according to claim 8 , in addition to the effect of the invention according to any one of claims 1 to 7 , the lower surface of the reagent cassette can be convex toward the reaction chamber instead of being horizontal. The reagent easily falls into the reaction chamber, and the reliability of the PCR is improved.

The invention according to claim 9 is a reagent cassette in which a reagent is stored and sealed in advance, and when a sample necessary for PCR is stored in a container and PCR is performed, heating of the PCR causes the sealing means to melt and the reagent The storage unit is opened. And, since the sample and the reagent move to the reaction chamber below the container, a desired amount and type of reagent can be stored in advance, and by using the container for PCR, the reliability and quickness of the PCR can be simply and stably. It is possible to improve the quality. In addition, since it becomes possible to seal the plurality of reagent storage units in a state in which the reagents stored in each are not mixed, it becomes easy to use a reagent that is not suitable for being mixed in advance. Furthermore, since only the sealing means is opened the reagent accommodating portion melted by heating P CR, stability of the reaction is above improvement.

The invention according to claim 10 , in addition to the effect of the invention according to any one of claims 1 to 9 , is a sealing means which is solid at normal temperature and which becomes liquid upon initial heating of PCR. Therefore, the certainty of PCR is improved.

According to an eleventh aspect of the invention, in addition to the effect of the invention according to any one of the first to tenth aspects, when sealing, from the upper edge of the partition wall to the upper open end surface of the basic skeleton Since the sealing means tends to stay at the location of the point, the reliability of the sealing is improved.

Claims (17)

A container for PCR,
A container body having a reaction chamber in the bottom of which a sample and a reagent are stored and reacted at the time of PCR;
A lid which can be fitted to the container body;
And a reagent cassette stored at a location between the lower surface of the lid and the reaction chamber inside the container body,
The reagent cassette has at least one reagent storage unit that can be sealed by a sealing unit that is solid before PCR and is melted by heating with PCR. The lower surface of the lid is formed with an upwardly recessed recess capable of storing at least a portion thereof by contact with a sample,
The PCR container according to claim 1, wherein the reagent storage portion has a shape penetrating in the vertical direction. In the reagent cassette, a plurality of the reagent storage units are provided,
The PCR container according to claim 1 or 2, wherein the plurality of reagent storage parts are separated from one another by a partition wall.   The reagent cassette according to any one of claims 1 to 3, wherein the reagent cassette has a cylindrical shape opened in the vertical direction, and a slit extending in the vertical direction and communicating with the reagent storage portion is formed on a side wall thereof. PCR container.   The PCR container according to any one of claims 1 to 4, wherein the reaction chamber is in the shape of a truncated pyramid and is made of a transparent material.   The container body, the lid, and the reagent cassette are set to have a dimensional relationship such that a gap is generated between the lower surface of the lid and the reagent cassette when the lid is fitted to the container body. The container for PCR according to any one of claims 1 to 5.   The container for PCR according to any one of claims 1 to 6, wherein at least a part of the inner surface of the container body protrudes inward toward the reagent cassette.   The PCR container according to any one of claims 1 to 7, wherein the container body has a thickness of 0.05 mm or more and 0.5 mm or less.   The container for PCR according to any one of claims 1 to 8, wherein the sealing means is wax, grease, paraffin or wax.   The container for PCR according to any one of claims 1 to 9, wherein the reagent cassette has an inwardly recessed concave portion formed on at least one of the upper surface and the lower surface. The lower surface of the lid is formed with an upwardly recessed recess capable of storing at least a portion thereof by contact with a sample,
The reagent storage portion is shaped to penetrate in the vertical direction,
An upwardly convex portion is formed on the upper surface of the reagent cassette,
The PCR container according to any one of claims 1 to 10, wherein the convex portion enters at least a part of the recess when the lid is fitted to the container body.   The PCR container according to any one of claims 1 to 11, wherein a downwardly convex portion is formed on the lower surface of the reagent cassette. A reagent-containing PCR container comprising the PCR container according to any one of claims 1 to 12,
The reagent-containing PCR container, wherein the reagent cassette contains the reagent in the reagent storage unit and is sealed by the sealing unit.   The container for PCR containing a reagent according to claim 13, wherein the reaction chamber is filled by the sealing means. A reagent cassette housed inside a PCR container,
Equipped with a reagent storage unit,
The reagent storage unit stores a reagent necessary for PCR and is sealed by a sealing unit which is solid before PCR and is melted by heating of PCR. A container for PCR,
The container includes a container body having a reaction chamber for storing and reacting a sample and a reagent at the bottom of PCR, and a lid which can be fitted to the container body.
The lid has at least one reagent storage formed at the tip through a portion of the exposed portion of the lower surface of the tip,
The other part of the lower surface of the tip part is formed with an upwardly recessed recess that can accommodate at least a part by contact with the sample,
The container for PCR, wherein the reagent storage portion is solid before PCR and the exposed portion can be sealed by a sealing unit that melts by heating of PCR.   The PCR container according to claim 16, wherein an opening penetrating the reagent storage portion is formed on a side wall of the tip portion.

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