WO2016148235A1

WO2016148235A1 - Container, kit for purifying nucleic acid, and method for manufacturing container

Info

Publication number: WO2016148235A1
Application number: PCT/JP2016/058492
Authority: WO
Inventors: 秀一明石
Original assignee: 凸版印刷株式会社
Priority date: 2015-03-18
Filing date: 2016-03-17
Publication date: 2016-09-22
Also published as: JPWO2016148235A1; JP6645493B2

Abstract

A container provided with a fluid removal part having: a recessed portion into which a dispensing device for dispensing a fluid can be inserted; a wiping member held inside the recessed portion, the wiping member being configured such that when the dispensing device is inserted into the recessed portion, the fluid that adheres to the outer surface of the dispensing device is removed; and a film deposited on both the open end of the recessed portion and the wiping member, the film covering the recessed portion.

Description

Container, nucleic acid purification kit, and container manufacturing method

The present invention relates to a container, a nucleic acid purification kit, and a method for producing the container.
This application claims priority based on Japanese Patent Application No. 2015-054972 filed in Japan on March 18, 2015, the contents of which are incorporated herein by reference.

With recent developments in genetic testing technology, for example, nucleic acid can be extracted from a sample such as a biological sample collected from a patient, and genetic differences such as single nucleotide polymorphisms can be detected to predict drug sensitivity in advance. Sex has been suggested.

When extracting nucleic acids from a specimen, reagent cartridges are used to extract the nucleic acids from the specimen in order to prevent cross-contamination between the specimens and to prevent the spread of infection sources. These reagents are used up and discarded together with the reagent cartridge every time the nucleic acid is extracted from the subject (see, for example, Patent Document 1).

Also, the liquid contained in the reagent cartridge is sucked and held by a dispensing tip having a cavity inside, and dispensed or stirred according to a predetermined process. At this time, the liquid may adhere to the outer surface of the dispensing tip in the step of sucking the liquid into the dispensing tip or the step of discharging the liquid from the dispensing tip. When the liquid adheres to the outer surface of the dispensing tip, there is a possibility that the accuracy of measuring the liquid using the dispensing tip may be reduced, or the contamination due to the liquid adhering to the outer surface of the dispensing tip may spread. For this reason, an automatic analyzer for analyzing nucleic acids is equipped with an apparatus for cleaning the outer surface of a dispensing tip (see, for example, Patent Document 2).

Furthermore, a sponge for wiping off the excess liquid adhering to the outer surface of the dispensing tip is accommodated in the reagent cartridge, and by inserting the dispensing tip into the cut of the sponge, the excess liquid adhering to the outer side of the dispensing tip is wiped off. A mechanism is provided in the reagent cartridge (see, for example, Patent Document 3).

International Publication No. 2005/118772 Japanese Patent Laid-Open No. 2008-215928 Japanese Unexamined Patent Publication No. 2011-072276

However, since the nucleic acid analyzer described in Patent Document 1 cannot remove the liquid adhering to the outer surface of the dispensing chip, a measurement error may occur due to the liquid adhering to the outer surface of the dispensing chip.
Moreover, in the automatic analyzer described in Patent Document 2, there is a problem that the apparatus configuration becomes complicated because the dispensing tip is vibrated to remove the cleaning liquid adhering to the outer surface of the dispensing tip.
In the method described in Patent Document 3, it is necessary to firmly fix the sponge to the reagent cartridge, and parts necessary for fixing are fitted and fixed, so that high precision management is required.
In addition, since the number of parts is large, assembly is very troublesome.

The present invention has been made in view of the above-described circumstances, and an object thereof is to provide a container, a nucleic acid purification kit, and a container manufacturing method that can remove liquid adhering to the outer surface of a dispensing chip with a simple configuration. That is.

The container according to the first aspect of the present invention includes a recess into which a dispensing device for dispensing a liquid can be inserted, and is held inside the recess, and the dispensing device is inserted into the recess when the dispensing device is inserted into the recess. A liquid removing unit comprising: a wiping member configured to remove the liquid adhering to the outer surface of the pouring device; and a film that is welded to both the opening end of the concave portion and the wiping member and covers the concave portion. Prepare.

In the first aspect described above, the melting point of the material of the wiping member may be higher than the melting point of the film.

In the first aspect, the wiping member may have lipophilicity.

In the first aspect, the wiping member may have hydrophilicity.

In the first aspect, the wiping member may be formed of a porous material having elasticity.

In the first aspect, the wiping member may be formed with a notch configured to be spread by the dispensing device.

In the first aspect, the wiping member may have a cut having an inner diameter that is smaller than a maximum outer shape of a region of the dispensing device that contacts the liquid.

In the first aspect, the liquid may be a nucleic acid separation and purification reagent.

In the first aspect, the dispensing device may be a dispensing device capable of quantitative dispensing.

The container according to the first aspect includes a sample storage unit that stores a sample containing nucleic acid, a liquid storage unit that stores the liquid, and a waste liquid storage unit that stores a waste liquid generated in the separation and purification of the sample. And an extraction filter cartridge for purifying the nucleic acid of the subject.

The nucleic acid purification kit according to the second aspect of the present invention includes the container according to the first aspect and a dispensing tip container for accommodating a plurality of dispensing tips that can be attached to a dispensing instrument.

In the container manufacturing method according to the third aspect of the present invention, a substantially bottomed cylindrical recess is formed, a wiping member into which a dispensing device can be inserted is inserted into the recess, and a heat-welding film is inserted into the recess. Of the opening end of the wiping member and a part of the upper surface of the wiping member, and after the film placement step, the entire circumference of the opening end and the portion of the upper surface of the wiping member that is in contact with the film. The opening end and the film are heat-welded by heating through the, and the wiping member and the film are heat-welded.

In the third aspect, when the opening end and the film are heat-welded and the wiping member and the film are heat-welded, the annular heating region following the contour shape of the opening end is used. The film may be heated to leave an unheated region surrounded by the heated region and including the center of the open end in an unwelded state.

In the third aspect, the wiping member may be a porous material having a through-hole-shaped cut along the center line of the recess.

According to the container and the nucleic acid purification kit according to the above aspect of the present invention, since the excess liquid is absorbed by the wiping portion in the liquid removal portion, the liquid adhering to the outer surface of the dispensing device can be removed with a simple configuration. Moreover, since the wiping member of the liquid removal part is welded by the film welded to the recessed part, a wiping member is hold | maintained at a recessed part through a film. For this reason, a wiping member can be hold | maintained in a recessed part with a simple structure.

It is a perspective view which shows the reagent cartridge which concerns on one Embodiment of this invention. It is a perspective view which shows the structure of the reagent cartridge and dispensing tip rack which concern on one Embodiment of this invention. It is sectional drawing which shows the structure of the extraction filter cartridge in the reagent cartridge which concerns on one Embodiment of this invention. It is sectional drawing which shows the structure of the liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. It is a disassembled perspective view which shows the structure of a part of liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. FIG. 4 is an enlarged view showing a part of a liquid removing unit in a reagent cartridge according to an embodiment of the present invention. (FIG. 4 is a plan view showing a configuration of a wiping filter in a liquid removing unit in a reagent cartridge according to an embodiment of the present invention. It is a top view which shows the structure of the wiping filter in the liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. It is a top view which shows the structure of the wiping filter in the liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. It is explanatory drawing for demonstrating the effect | action of the liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. It is explanatory drawing for demonstrating the effect | action of the liquid removal part in the reagent cartridge which concerns on one Embodiment of this invention. It is a figure for demonstrating the manufacturing method of the reagent cartridge which concerns on one Embodiment of this invention, and the process of wiping a sealing film and welding to a filter and a recessed part is shown. It is a figure for demonstrating the manufacturing method of the reagent cartridge which concerns on one Embodiment of this invention, and the process of wiping a sealing film and welding to a filter and a recessed part is shown. It is a figure for demonstrating the manufacturing method of the reagent cartridge which concerns on one Embodiment of this invention, and the process of wiping a sealing film and welding to a filter and a recessed part is shown. It is a figure for demonstrating the manufacturing method of the reagent cartridge which concerns on one Embodiment of this invention, and the process of wiping a sealing film and welding to a filter and a recessed part is shown. It is a top view of the reagent cartridge which concerns on one Embodiment of this invention, and has shown the welding area | region of the sealing film with respect to a wiping filter and a recessed part. It is sectional drawing in the AA of FIG. 8A.

Hereinafter, a container and a nucleic acid purification kit according to an embodiment of the present invention will be described with reference to FIGS.
FIG. 1 is a perspective view showing a reagent cartridge 100 which is an embodiment of the container of the present invention. FIG. 2 is a perspective view showing the configuration of the reagent cartridge 100 and the dispensing tip rack 200. FIG. 3 is a cross-sectional view showing the configuration of the extraction filter cartridge 150 in the reagent cartridge 100. 4A is a cross-sectional view showing the configuration of the liquid removing unit 128 in the reagent cartridge 100, and FIG. 4B is an exploded perspective view showing the configuration of a part of the liquid removing unit 128. FIG. 4C is an enlarged view of the liquid removing unit 128. 5A to 5C are plan views showing the configuration of the wiping filter 129A in the liquid removing unit 128. FIG. 6A and 6B are explanatory diagrams for explaining the operation of the liquid removing unit 128 in the reagent cartridge 100. FIG.

As shown in FIG. 2, the nucleic acid purification kit 10 includes a reagent cartridge 100 containing a reagent for extracting nucleic acid from a subject and a plurality of dispensing chips 201 for dispensing a liquid. An injection chip rack (dispensing chip container) 200 is provided. In the present embodiment, the dispensing tip rack 200 includes a plurality of dispensing tips 201 having the same shape and the same size. The liquid stored in the reagent cartridge 100 is dispensed or stirred by any one of the plurality of dispensing tips 201 so that no cross contamination occurs between the liquids by the dispensing tips 201. The dispensing tip rack 200 is also a container for collecting the dispensing tips 201 after use. After the use of the dispensing tip 201 in the nucleic acid analyzer, the dispensing tip 201 can be discarded as the infectious waste together with the dispensing tip rack 200.

The reagent cartridge 100 includes a main body 101 formed in a substantially box shape and a claw portion 102 formed so as to protrude from the outer surface of the main body 101. For example, when the reagent cartridge 100 is set in an automatic analyzer or the like, the claw portion 102 is configured to be able to engage with a part of the automatic analyzer so that the reagent cartridge 100 does not fall down.
A part of the outer surface of the main body 101 is wound with a thin-film sealing film 103 that is removed when the nucleic acid purification kit 10 is used. The opening of the main body 101 is sealed by the sealing film 103, so that an extraction filter cartridge 150, which will be described later, disposed inside the main body 101 does not fall from the main body 101 and foreign substances such as dust are present inside the main body 101. It can be prevented from mixing.

Inside the main body 101, a sample well (subject storage part) 110, a reagent well part 120, a waste liquid well (waste liquid storage part) 130, and a recovery well 140 are integrally formed. A sample such as a biological sample is placed in the sample well 110. The reagent well section 120 contains a reagent for extracting nucleic acid from the subject. The unnecessary solution separated in the step of extracting nucleic acid from the subject is discarded in the waste well 130. The recovery well 140 recovers the nucleic acid extracted from the subject.

The reagent cartridge 100 has an extraction filter cartridge 150 containing a carrier that adsorbs nucleic acids. The reagent cartridge 100 is integrally formed with a holding portion 160 in which the extraction filter cartridge 150 is accommodated.

The reagent well section 120 includes a plurality of reagent wells (reagent storage sections) 121, 122, 123, 124, 125, 126, an oil well (oil storage section) 127, and an oil removal section (liquid removal section) 128. is doing. In the reagent well portion 120, the openings of the plurality of

reagent wells

121, 122, 123, 124, 125, 126, and the oil well 127, and the opening of the oil removal portion 128 are shown in FIGS. 1, 4A, and 4C. It is sealed with a sealing film 104.

The sealing film 104 is preferably configured such that gas permeation is suppressed and the film can be broken by piercing the dispensing tip 201. For example, a metal thin film or a plastic film is used. Can do. As the sealing film 104, a film in which metal and plastic are combined can also be used.
As shown in FIG. 4C, the sealing film 104 of this embodiment is a laminate having a sealing layer (an aluminum layer 104A formed of aluminum in this embodiment), an adhesive layer 104B, and a welding layer 104C. is there. The sealing layer prevents liquid and gas permeation. The adhesive layer 104B is provided on the aluminum layer 104A. The weld layer 104 C can be thermally welded to the plurality of

reagent wells

121, 122, 123, 124, 125, 126 and the opening ends of the oil well 127 and the opening end of the oil removing unit 128.

The aluminum layer 104A is provided for improved tearability and a water vapor barrier. Examples of the material of the aluminum layer 104A include, but are not limited to, thin film metals and films.

The melting temperature of the welding layer 104 C of the sealing film 104 is preferably set to the same level as the melting temperature of the main body 101. The melting temperature of the wiping filter 129 A is preferably set to be higher than both the melting temperature of the welding layer 104 C of the sealing film 104 and the melting temperature of the main body 101. By setting such a melting temperature, the wiping filter 129 A is hardly melted at the time of thermal welding using the sealing film 104. In addition, since the wiping filter 129A is difficult to melt, the wiping filter 129A maintains a porosity capable of absorbing excess liquid adhering to the dispensing tip 201 described later.

The welded layer 104C and the aluminum layer 104A are bonded by an adhesive layer 104B. The adhesive layer 104B is provided to closely bond the weld layer 104C and the aluminum layer 104A, and any type can be used as long as it is an adhesive that can bond both.

The cut film 105 is formed in the sealing film 104. In order to smoothly insert the dispensing tip 201 into the wiping filter 129 A described later, a cut is formed in advance when the sealing film 104 is bonded.

In the reagent wells 121 to 126 shown in FIG. 2, a lysis solution 121A that dissolves a biological material such as a cell membrane, and a lysis that dissolves a biological material such as cytoplasm that cannot be dissolved by the lysis solution 121A and clogs the carrier. Liquid 122A, washing liquids 123A and 124A for washing away unnecessary substances other than the nucleic acid adsorbed on the carrier, elution liquid 125A for eluting nucleic acid from the carrier, and diluting liquid 126A for adjusting the nucleic acid concentration in the elution liquid, respectively. It is housed individually.
In the usage mode described later, the analysis reagent is arranged on the nucleic acid analysis chip. Alternatively, the analysis reagent may be stored in a reagent well. For example, analysis reagent premixes in which a part of reagents for performing PCR and SNP measurement by Invader (registered trademark) method on nucleic acids are mixed in advance can be individually accommodated in each reagent well.

In the oil well 127, for example, a well-known oil 127A used in a PCR reaction in a layered manner is accommodated. As the oil 127A, for example, mineral oil or silicon oil can be preferably used.

As shown in FIGS. 4A to 4C, the oil removing unit 128 holds the wiping unit 129 inside the wiping unit 129 for removing the oil 127A adhering to the outer surface of the dispensing tip 201 (see FIG. 2). And a recess 128A.

As shown in FIGS. 4A and 4B, the wiping portion 129 has a wiping filter 129A having lipophilicity. The wiping filter 129A having lipophilicity can satisfactorily remove oily liquids such as mineral oil. Note that in the case where a hydrophilic wiping filter is provided instead of the lipophilic wiping filter 129A, the aqueous solution can be satisfactorily removed. The wiping filter 129 A is formed in a substantially cylindrical shape or a substantially disk shape along the inner diameter of the oil removing portion 128. In the center of the wiping filter 129A, a cut portion 129C formed so as to penetrate in the central axis direction of the wiping filter 129A is provided.
The wiping filter 129A is preferably formed of a porous member having elasticity. For example, a wiping filter formed by compressing a sponge filter or the like can be used. The main thing to be used includes rubber sponge, urethane sponge, polyethylene foam, EVA (Ethylence Vinyl Acetate) sponge, and the like.

As shown in FIG. 5A, the cut portion 129C is located in the center of the wiping filter 129A when the wiping filter 129A is viewed in the central axis direction, and is formed in a cross shape. As shown in FIG. 5B, instead of the cut portion 129C, the wiper filter 129A may have a cut portion 129D formed in a true circle when viewed in the central axis direction. Further, as shown in FIG. 5C, instead of the cut portion 129 C, a circular cut portion 129 E having a bulging portion that bulges radially inward of the wiping filter 129 A may be provided. The inner diameter of the cut portion is formed so as to be smaller than the maximum outer shape of the area of the dispensing instrument that comes into contact with the oil.

Further, in the oil removing portion 128, an annular region 128 B (hereinafter sometimes referred to as a flange portion) surrounding the center of the opening of the recess 128 A protrudes from the surface of the main body 101 where the reagent well portion 120 is formed. It is high. The sealing film 104 is fixed to a portion of the oil removing portion 128 that protrudes from the surface where the reagent well portion 120 is formed.

As shown in FIG. 2, the waste liquid well 130 is a recess formed along the outer diameter shape of the extraction filter cartridge 150 and has a shape capable of supporting the extraction filter cartridge 150. In the state where the extraction filter cartridge 150 is attached to the waste well 130, the extraction filter cartridge 150 does not fall within the reagent cartridge 100.

The recovery well 140 can support the extraction filter cartridge 150 similarly to the waste liquid well 130. The bottom of the recovery well 140 has a container shape that can store the nucleic acid solution eluted from the carrier of the extraction filter cartridge 150 by the eluent 125A.

The waste liquid well 130 and the recovery well 140 are provided adjacent to each other in the reagent cartridge 100. This is to shorten the flow line of the extraction filter cartridge 150 when the extraction filter cartridge 150 is moved to the recovery well 140 after the extraction filter cartridge 150 is washed in the waste liquid well 130. Thereby, the possibility that the extraction filter cartridge 150 passing over the reagent cartridge 100 contaminates the reagent cartridge 100 or the like can be reduced.

As shown in FIG. 3, the extraction filter cartridge 150 has a substantially cylindrical main body 151 as an outer frame of the extraction filter cartridge, and an extraction filter unit 152 provided inside the main body 151.

The main body 151 has an upper opening 151A and a lower end 151B that are both open. Further, the main body 151 is formed in a funnel shape having an opening diameter smaller than the opening of the upper opening portion 151A on the lower end 151B side with respect to the extraction filter unit 152. The lower end 151B is provided with a nozzle-like discharge port 151C protruding downward.
To the opening of the upper opening 151A, the

lysing solutions

121A and 122A, the cleaning solutions 123A and 124A, the eluent 125A, and the like in a state where the specimen is dissolved are supplied. These liquids pass through the filter unit 152 and are discharged from the outlet 151C.

The filter unit 152 includes an adsorption filter 152A, a support member 152B, and a washer 152C. The adsorption filter 152A contains a carrier having the property of adsorbing nucleic acids. The support member 152B is disposed closer to the lower end 151B than the adsorption filter 152A and prevents the adsorption filter 152A from being deformed. The washer 152C fixes the adsorption filter 152A.

The adsorption filter 152A is formed in a film shape by a porous material that can adsorb nucleic acid. The adsorption filter 152A has a disk shape. The shape of the adsorption filter 152 is not limited to the disk shape, and a suitable shape may be selected in accordance with the shape of the main body 151 of the extraction filter cartridge 150. The material of the adsorption filter 152A is preferably a material having a property that the nucleic acid is adsorbed in the cleaning liquids 123A and 124A, and the adsorbed state of the nucleic acid is weakened in the eluate 125A. The adsorption filter 152A is preferably a porous material in which a hydroxyl group is introduced as a hydrophilic group. Specifically, the adsorption filter 152A is formed of silica or by combining silica on another substance. The material of the adsorption filter 152A is not particularly limited as long as it is a material that can adsorb a biological substance in the presence of an organic substance. Further, the adsorption filter 152A may be formed to have porosity by overlapping and forming fiber materials such as glass wool.

The support member 152B is preferably formed of a material that has at least low adsorptivity to nucleic acids and does not inhibit the reaction of extracting nucleic acids from the specimen. For example, the support member 152B can be formed by baking and hardening resin particles. The rigidity of the support member 152B is higher than that of the adsorption filter 152A, and the support member 152B prevents the adsorption filter 152A from being deformed in the main body 151.

The washer 152C is preferably formed of a material that has at least low adsorptivity to nucleic acid and does not inhibit the reaction of extracting nucleic acid from the specimen. The washer 152C is installed so as to be in contact with the upper surface of the adsorption filter 152A, and is engaged with and fixed to the inside of the main body 151. The washer 152 C suppresses the generation of a gap by contacting the suction filter 152 A in a circumferential shape.

As shown in FIG. 2, the holding unit 160 is in the initial position in the reagent cartridge 100 where the extraction filter cartridge 150 is accommodated. In addition, an absorber (not shown) that absorbs liquid can be provided on the bottom of the holding unit 160. This absorber contacts the outer surface of the extraction filter cartridge 150 on the side of the outlet 151 C when the extraction filter cartridge 150 is accommodated in the holding unit 160. For this reason, for example, when the cleaning liquid 123A adheres to the outer surface of the outlet 151C when the cleaning liquid 123A is supplied into the extraction filter cartridge 150, the cleaning liquid 123A can be removed by absorbing the cleaning liquid 123A into the absorber.

The operation of the reagent cartridge 100 and the nucleic acid purification kit 10 of the present embodiment having the above-described configuration will be described focusing on the operation of the oil removal unit (liquid removal unit) 128. Hereinafter, as an automatic analyzer to which the nucleic acid purification kit 10 is applied, an automatic analyzer that has a dispensing conveyance mechanism that conveys the dispensing chip 201 and performs PCR analysis after purifying nucleic acid will be described as an example. explain.

First, the sealing film 103 of the reagent cartridge 100 shown in FIG. 1 is removed manually by the user. Subsequently, for example, a whole blood sample is injected into the sample well 110 of the reagent cartridge 100 manually by the user.
The reagent cartridge 100 containing the sample and the dispensing tip rack 200 are respectively installed at predetermined locations of the automatic analyzer.

Subsequently, various reagents stored in the reagent wells 121 to 126 are dispensed and mixed by the dispensing transport mechanism of the automatic analyzer according to a predetermined procedure. Thereby, the cells in the whole blood sample supplied to the sample well 110 are lysed to obtain a cell lysate. When the liquid is sucked into the dispensing tip 201 from the reagent wells 121 to 126, the tip of the dispensing tip 201 is inserted into the sealing film 104 that seals the reagent wells 121 to 126. Then, a through-hole is formed in the sealing film 104, and various reagents in the reagent wells 121 to 126 can be sucked by the dispensing tip 201.
The dispensing tip 201 may be automatically replaced depending on the reagent used and the work process.

First, since it is necessary to collect the waste liquid, the extraction filter cartridge 150 is transported to the waste liquid well 130. Next, a predetermined amount of blood is dispensed from the sample well 110 to the reagent well 121. The reagent well 121 contains a lysing solution 121A, and pipetting (operation of repeating suction and discharge of the solution with respect to the dispensing tip) is performed in the reagent well 121 to lyse cells in the blood. Then, the solution in which the cells are lysed is supplied from the reagent well 121 to the extraction filter cartridge 150 placed in the waste liquid well 130. The extraction filter cartridge 150 can increase the speed at which the liquid passes through the adsorption filter 152A by feeding gas from the upper opening 151A and pressurizing the main body 151. Then, the solution in which the cells are dissolved passes through the adsorption filter 152A, and the nucleic acid is adsorbed on the adsorption filter 152A. Thereafter, the adsorption filter 152A is washed with a dissolving solution 122A that dissolves biological substances such as cytoplasm that cannot be completely dissolved by the dissolving solution 121A and clog the carrier.

Further, the cleaning liquids 123A and 124A are supplied to the adsorption filter 152A, and the adsorption filter 152A is cleaned with the cleaning liquids 123A and 124A.
A porous material that absorbs liquid, such as a sponge, can be installed at the bottom of the waste liquid well 130. By using a material that absorbs the dissolution liquid, the cleaning liquid, and the like, these liquids can be held in the reagent cartridge, and these liquids can be prevented from coming out of the reagent cartridge 100.
Thereafter, the extraction filter cartridge 150 is transported to the recovery well 140, and the eluate 125A is supplied to the adsorption filter 152A. Thereby, the nucleic acid adsorbed by the adsorption filter 152A is eluted in the eluent 125A, and the nucleic acid solution containing the nucleic acid is recovered in the recovery well 140.

Further, the diluent 126A and the eluate 125A from which the recovered nucleic acid is recovered are mixed together, and the sample preparation is completed.
The nucleic acid separation and purification by the nucleic acid purification kit 10 is thus completed.

Subsequently, PCR is performed on the nucleic acid using the nucleic acid separated and purified by the nucleic acid purification kit 10. The automatic analyzer sucks the nucleic acid solution with the dispensing chip 201 and supplies the nucleic acid solution to a predetermined reaction container (for example, a microtube). Subsequently, the dispensing tip 201 is inserted into the oil well 127 of the nucleic acid purification kit 10 shown in FIG. 1, and the oil 127 A accommodated in the oil well 127 is sucked into the dispensing tip 201. At this time, the oil 127A may adhere to the outer surface of the dispensing tip 201 in the form of droplets because the material of the dispensing tip 201 has a high affinity with the oil 127A.

The dispensing transport mechanism of the automatic analyzer moves the dispensing tip 201 to the oil removing unit 128 as shown in FIG. 6A. Further, as shown in FIG. 6B, the dispensing transport mechanism of the automatic analyzer inserts the tip 201A of the dispensing tip 201 through the sealing film 104 into the cut portion 129C of the wiping filter 129A of the oil removing unit 128. To do. The sealing film 104 and the wiping filter 129A are fixed to each other by heat welding. Therefore, even if the wiping filter 129A is pushed toward the bottom of the recess 128A of the oil removing unit 128 by the pushing force of the dispensing tip 201 to the wiping filter 129A, the wiping filter 129A is held without being peeled off from the sealing film 104. Is done.

The oil 127A adhering to the outer peripheral surface of the tip 201A of the dispensing tip 201 is sucked by the wiping filter 129A and removed from the outer peripheral surface of the dispensing tip 201. By pulling out the dispensing tip 201 from the cut portion 129C, the outer peripheral surface of the dispensing tip 201 comes into contact with the wiping filter 129A again, and the oil 127A attached to the outer peripheral surface of the dispensing tip 201 is wiped off.

The oil 127A is superposed on the nucleic acid solution in the reaction vessel described above by the dispensing tip 201 in which the oil 127A is held and the oil 127A on the outer peripheral surface is removed. At this time, since the excess oil on the outer surface of the dispensing tip 201 is removed, the oil does not adhere to the wall surface of the reaction vessel.
By superposing the oil 127A on the nucleic acid solution in the reaction vessel, the nucleic acid solution is prevented from evaporating in the PCR reaction, and the PCR reaction can be suitably performed.

Next, the manufacturing method of the reagent cartridge 100 (container) of the above embodiment will be described focusing on the process of welding the sealing film 104 to the oil removing unit 128 having the wiping filter 129A. FIG. 7A to FIG. 7D are views for explaining the method of manufacturing the reagent cartridge according to this embodiment, and show a process of wiping the sealing film and welding it to the filter and the recess. FIG. 8A is a plan view of the reagent cartridge according to the present embodiment, and shows a welding region of the sealing film to the wiping filter and the recess. FIG. 8B is a cross-sectional view taken along line AA in FIG. 8A.

As shown in FIG. 7A, in the present embodiment, the wiping filter 129A is inserted into the recess 128A of the oil removing unit 128. The wiping filter 129A has a height that slightly protrudes from the recess 128A.
As shown in FIG. 7B, the sealing film 104 is disposed so as to overlap the upper surface of the wiping filter 129A attached to the recess 128A so as to protrude slightly from the recess 128A. 4C and 7C, while the sealing film 104 is heated, the welding layer 104C of the sealing film 104 is pressed against the opening portion of the recess 128A. The heat bar 300 can be heated to a temperature exceeding the melting temperature of the welding layer 104 C of the sealing film 104.
When the sealing film 104 is bonded to the reagent well portion 120 of the reagent cartridge 100 by heat, the wiping filter 129 A is installed so as to come into contact with the welding layer 104 C of the sealing film 104. As a result, as shown in FIG. 4C, the welding layer 104C of the sealing film 104 melts and soaks into the porous material of the wiping filter 129A to form an adhesive layer 104D after heating. As shown in FIG. 7D, the post-heating adhesive layer 104 D is a layer generated so as to firmly bond the sealing film 104 and the wiping filter 129 A when the sealing film 104 is heated.
In the present embodiment, an annular region 128B (hereinafter sometimes referred to as a flange portion) surrounding the center of the opening of the recess 128A is higher than the surface on which the reagent well portion 120 is formed. For this reason, when the sealing film 104 is adhered, the heating portion is reduced, and generation of wrinkles or damage of the sealing film 104 can be prevented.
In addition, although it is preferable that the oil removal part 128 has the flange part 128B, the oil removal part 128 does not need to have the flange part 128B.

As shown in FIG. 8A, the contact region 301 of the heat bar 300 (see FIG. 7C) with respect to the sealing film 104 is preferably an annular region 128B surrounding the center of the opening of the recess 128A. When the heat bar 300 is in contact with the annular region 128B, as shown in FIG. 8B, the sealing film 104 and the wiping filter 129A are in an unwelded or weakly welded state near the center of the opening of the recess 128A. The vicinity of the center of the opening of the recess 128A is an area where the dispensing tip 201 is assumed to be inserted, and in this embodiment, a cut portion 129C is arranged as shown in FIGS. 8A and 8B. Since the region where the cut portion 129C is arranged is not welded or is weakly welded, a force required to penetrate the sealing film 104 for inserting the dispensing tip 201 into the cut portion 129C can be reduced. Furthermore, since the region near the center of the opening of the recess 128A is not welded or weakly welded, the portion of the wiping filter 129A near the center of the opening of the recess 128A has a weld layer 104C that becomes the adhesive layer 104D after heating. There are few components (refer FIG. 8B). For this reason, the absorptivity with respect to the oil etc. used as an excess liquid does not fall easily. Note that the weld layer 104C may be uniformly formed on the wiping filter 129A.

As described above, according to the reagent cartridge 100 and the nucleic acid purification kit 10 according to the present embodiment, the oil 127A is absorbed by the filter 129A in the oil removing unit 128 provided in the reagent cartridge 100. Therefore, the oil 127A attached to the outer surface of the dispensing tip 201 can be suitably removed with a simple configuration.
Further, the wiping filter 129 A of the oil removing unit 128 is welded to the sealing film 104 welded to the recess 128 A of the oil removing unit 128. For this reason, the wiping filter 129 A is held in the recess 128 A via the sealing film 104. Thus, in the present embodiment, the wiping filter 129A can be held in the recess 128A with a simpler configuration than the conventional one.

In addition, since the nucleic acid purification kit 10 includes the oil removing unit 128, the oil 127A attached to the outer peripheral surface of the tip 201A of the dispensing tip 201 can be removed by the wiping filter 129A of the oil removing unit 128. It is possible to prevent the liquid from unintentionally adhering to the wall of the reaction vessel and the like, and to improve the accuracy and reproducibility when examining nucleic acids.

Moreover, since the wiping filter 129A has elasticity, the oil 127A attached to the outer surface of the dispensing tip 201 can be wiped off by pressing the wiping filter 129A against the outer surface of the dispensing tip 201. Furthermore, since the wiping filter 129A is formed of a porous material, the oil 127A can be suitably absorbed and held in the wiping filter 129A. For this reason, even if the wiping filter 129A is used a plurality of times, the oil 127A can be suitably absorbed.

Also, the reagent and filter unit necessary for extracting the nucleic acid and the oil removing unit 128 are provided inside the reagent cartridge 100 and integrated as a kit. Therefore, since only the operation of adding the specimen to the sample well 110 and setting it in the automatic analyzer is performed manually, the genetic test can be performed easily. Further, since the waste liquid well 130 is integrally provided in the reagent cartridge 100, the reagent cartridge 100 may be simply removed from the automatic analyzer and discarded after the genetic test is completed. For this reason, waste liquid processing is simple and there is no possibility that the surroundings will be contaminated by residual liquid such as a subject.

Next, the present invention will be described with reference to examples.
(Example 1)
The sealing film 104 is welded to the wiping filter 129A, and the reagent cartridge 100 of this embodiment that holds the wiping filter 129A in the recess 128A, and the sealing film 104 is wiped without being welded to the wiping filter 129A. The case where the filter 129A was arranged as disclosed in Patent Document 3 was compared.
Both the reagent cartridge 100 of this example and the technique disclosed in Patent Document 3 were able to remove the excess liquid (oil) using the wiping filter 129A satisfactorily.

(Example 2)
The example which welded with respect to the flange part 128B of 128 A of recessed parts is shown regarding the area | region where the sealing film 104 is wiped and adhered to the filter 129A. In the present embodiment, the region near the center of the opening of the recess 128A is in a welded state weaker than the state in which the heat bar 300 is in contact because the heat bar 300 is not in contact. This difference in the welded state is reflected in the difference in force required for penetrating the sealing film 104 using the dispensing tip 201. In the portion near the center of the opening of the recess 128A, it is possible to penetrate the sealing film 104 with a force of 4N, and in the annular region surrounding the center of the opening of the recess 128A, 12N to penetrate the sealing film 104. The power of was necessary.

As mentioned above, although embodiment of this invention was explained in full detail with reference to drawings, the concrete structure is not restricted to this embodiment, The design change etc. of the range which does not deviate from the summary of this invention are included.
For example, in the above-described embodiment, an example in which the oil removing unit (liquid removing unit) 128 removes the oil attached to the outer surface of the dispensing tip 201 has been described. Moreover, the liquid adhering to the outer surface of the dispensing tip 201 can be suitably removed. For example, the liquid removal unit of the present invention can be suitably applied when operating glycerol, a surfactant, or the like with high viscosity.

10 Nucleic acid purification kit 100 Reagent cartridge (container)
104 Sealing film (film)

104A Aluminum layer

104B Adhesive layer 104C Welded layer 104D Adhesive layer after heating 128 Oil removal part (liquid removal part)
128A Concave part 129 Wiping part 129A Wiping filter (wiping member)
129C Notch 200 Dispensing tip rack (dispensing tip container)
201 Dispensing tip (dispensing instrument)

Claims

A recess into which a dispensing device for dispensing liquid can be inserted;
A wiping member held inside the recess and configured to remove the liquid adhering to the outer surface of the dispensing instrument when the dispensing instrument is inserted into the recess;
A container provided with a liquid removal unit having a film that is welded to both the opening end of the recess and the wiping member and covers the recess.
The container according to claim 1, wherein a melting point of the material of the wiping member is higher than a melting point of the film.
The container according to claim 1, wherein the wiping member has lipophilicity.
The container according to claim 1, wherein the wiping member has hydrophilicity.
The container according to claim 1, wherein the wiping member is formed of a porous material having elasticity.
The container according to claim 1, wherein the wiping member is formed with a notch configured to be spread by the dispensing device.
The container according to claim 1, wherein the wiping member has an incision having an inner diameter smaller than a maximum outer shape of a region of the dispensing device that contacts the liquid.
The container according to claim 1, wherein the liquid is a reagent for nucleic acid separation and purification.
The container according to claim 1, wherein the dispensing device is a dispensing device capable of quantitative dispensing.
A specimen container that contains a specimen containing nucleic acid;
A liquid container for containing the liquid;
A waste liquid storage unit for storing a waste liquid generated in the separation and purification of the analyte;
An extraction filter cartridge for purifying the nucleic acid of the analyte;
The container according to any one of claims 1 to 9, further comprising:
A container according to any one of claims 1 to 9;
A dispensing tip container for housing a plurality of dispensing tips attachable to the dispensing instrument;
A nucleic acid purification kit comprising:
Forming a substantially bottomed cylindrical recess,
Insert a wiping member into which the dispensing device can be inserted into the recess,
A heat-welding film is placed on the opening end of the recess and part of the upper surface of the wiping member,
By heating the entire circumference of the open end and the portion of the upper surface of the wiping member in contact with the film via the film, the open end and the film are thermally welded, and the wiping member A method for producing a container for thermally welding the film.
When the opening end and the film are heat-welded and the wiping member and the film are heat-welded, the film is heated against an annular heating region that follows the contour shape of the opening end, The manufacturing method of the container of Claim 12 which leaves the non-heating area | region which is surrounded by a heating area | region and contains the center of the said opening end in an unwelded state.
The method for producing a container according to claim 13, wherein the wiping member is a porous material having a through-hole-like cut along the center line of the recess.