JP2015195784A - Porous filter column, reagent cartridge, and nucleic acid extraction kit - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a porous filter column capable of maintaining solution permeability and a moving amount of fluid at high levels, and can be easily and securely assembled regardless of an operator while securely holding a porous filter in a column without a gap and without creating a crack.SOLUTION: A filter column comprises: an upper container 11; a lower container 12 bonded to a lower part of the upper container 11; and a porous filter 21 and a support member 22 which are arranged inside the lower container 12, in which a filter press/bonding part 1112 of the upper container 11 is fitted into a cylindrical part 1211 of the lower container 12 to bond the upper container 11 and the lower container 12. The porous filter 21 is held inside the column by pressing force from the filter press/bonding part 1112 which is obtained by engaging a locking part of the filter press/bonding part 1112 with a locking part 1214 provided for the cylindrical part 1211 of the lower container 12 when the upper container 11 and the lower container 12 are bonded.

Description

The present invention, filtration or liquids, porous filter column simple prefabricated used such as extraction of a specific substance in a liquid, more particularly, a porous filter column characterized by a holding mechanism of the porous filter and, The present invention relates to a reagent cartridge and a nucleic acid extraction kit using the porous filter column.

  Porous filters are widely used in research and industrial applications as tools for filtering liquids and extracting specific substances contained in liquids. In such cases, the porous filter must be held in a column through which the liquid passes. Usually, a method in which a porous filter is sandwiched between two members in a column is used. In recent years, such a porous filter column is sometimes used for nucleic acid extraction of a biological sample in the fields of genetic engineering and genetic medicine.

  The BOOM method is known as a nucleic acid extraction / recovery method. The BOOM method is a method for separating and purifying nucleic acid using a combination of a chaotropic reagent and solid phase silica utilizing the fact that nucleic acid is adsorbed on the silica surface in the presence of chaotropic ions. This is a technique for separating and purifying nucleic acids by adsorbing them on a high-quality silica, washing away impurities with a washing solution, and then eluting and collecting the nucleic acids adsorbed on the silica with an eluent. In order to carry out this technique, a bottomed cylindrical column having a porous filter held at the bottom is used. These have a waste liquid outlet at the bottom of a bottomed cylindrical column, and pass through a porous filter by a pressurizing means such as a pump or a centrifuge to collect the finally purified nucleic acid. Is.

  The process of purifying nucleic acid from human blood using a pressurizing means such as a pump or centrifuge will be explained more specifically. (1) Dissolving blood → (2) Moving the dissolved blood to the upper part of the porous filter Then, the nucleic acid is adsorbed → (3) Cleaning of porous filter → (4) Drying of porous filter → (5) Recovery of nucleic acid from porous filter.

  In the steps (2) to (5), it is necessary to apply pressure from the air layer to the porous filter. Here, for example, in the step (2), when there is a problem in holding the porous filter in the column through which the liquid passes and there is a gap between the end of the porous filter and the column, the dissolved blood Part of the liquid flows out of the gap or the like without passing through the porous filter.

  In addition, when a gap is generated between the end of the porous filter and the column in the step (4), the pressure due to the air layer is not uniformly applied to the front surface of the porous filter, and the air flows out from the gap or the like. It will be insufficiently dried. As a result, ethanol or the like contained in the cleaning liquid used in the step (3) remains, and when nucleic acid is collected from the porous filter, ethanol or the like is mixed into the collected nucleic acid. If such contamination is present, the occurrence rate of defects related to the reaction using the collected nucleic acid increases.

  In the porous filter column for nucleic acid extraction, most of the members for holding the cylindrical outer container and the porous filter are made of resin and used for precise experiments and measurements. In order to prevent, once used, it is often thrown away without being used again. In addition, the cylindrical outer container and the porous filter are handled as a unit, and a porous filter holding member for eliminating a gap between the end of the porous filter and the column is required.

An O-ring is generally used as a member for holding the porous filter. In a bottomed cylindrical column that supports the porous filter, the O-ring is installed at the outer peripheral edge of the porous filter. There is something that is.
However, since the filter portion covered with the O-ring cannot be used for filtration, the filter area available for filtration becomes narrow, and the filtration efficiency decreases. As a result, the amount of fluid movement is unlikely to increase even when pressurized. In addition, since the solution is likely to remain in the gap formed between the O-ring and the column, both the adsorption efficiency and the washing efficiency to the porous filter are poor in nucleic acid extraction applications, and the purity and yield of the sample are reduced. It was.

  With respect to this problem, Patent Document 1 describes a filter holding member that holds the outer peripheral edge of a porous filter over the entire circumference. Since this member can crush the outer peripheral portion uniformly, it is difficult to generate a gap, and as a result, pressure leakage or the like hardly occurs. However, since it is a fit, the accuracy of the diameter is required, and in order to avoid the same problems as those of the O-ring, the manufacturing method is considerably limited because the thickness is reduced, resulting in an expensive part.

  Also, in the process of fitting the filter holding member at the time of assembly, it is ideal to fit it by pushing in a certain amount, but since there are variations in the dimensions of the filter holding member and filter column, push it to the point where it can be pushed. It is made to fit by the method. However, when the filter member is flexible, the filter member is crushed by being pushed in and the portion directly below the filter holding member is crushed. As a result, the center portion is pulled and the filter member may be torn.

  With respect to this problem, Patent Document 2 describes a manufacturing method and a shape in which a filter is changed to a hard mold, and the filter is mounted on a primary molded product and then installed on the mold, and integrally molded. . In this method, the filter member and the filter holding member can be easily brought into close contact with each other to obtain good adhesion, and the generation of a gap can be minimized. However, if the filter is installed in the mold and molded integrally, there is a high possibility that unnecessary heat will be applied to the filter and it will be disadvantageous for the nucleic acid extraction ability, and if the filter member is soft like glass fiber, injection The filter cannot be fixed by the pressure at the time of molding, and the shape as designed cannot be obtained.

Japanese Patent Application No. 2012-173417 JP 2010-94136 A

  The present invention solves the above-mentioned problems, and while maintaining the porous filter in the column reliably, the permeability of the solution and the amount of movement of the fluid are maintained high, no gap is generated, and no heat is applied. A porous filter column that can be easily assembled even with a soft filter, does not crack, and can be assembled easily and reliably without selecting an operator, and a reagent cartridge and a nucleic acid extraction kit using the same The purpose is to provide.

  In order to achieve the above object, the invention of claim 1 is a porous filter column, which is disposed in a cylindrical upper container, a lower container coupled to the lower part of the upper container, and the lower container. A support member, and a porous filter that is supported by the support member in the lower container and disposed over the entire surface perpendicular to the axial direction of the upper container, the upper container having an opening at the upper end. A cylindrical portion having a cylindrical portion having a cylindrical shape, and a coupling portion for a filter presser combined with a cylindrical shape provided at a lower portion of the outer peripheral surface of the cylindrical portion, and the lower container is closely fitted to an outer periphery of the combined portion for combined filter presser And a bottom surface part connected to the lower part of the cylindrical part and formed with a discharge port, and a first locking part is formed on the outer peripheral surface of the filter pressing and coupling part, and the inner part of the cylindrical part of the lower container The peripheral surface is engaged with the first locking portion and the front A second locking portion for defining a fitting depth of the coupling part for the filter presser to the cylindrical part of the lower container is formed, and the coupling part for the filter presser combined with the cylindrical part of the lower container is fitted to the first cylindrical part. The lower end of the filter pressing / combining portion is configured to contact the upper surface of the peripheral edge of the porous filter with a constant pressing force in a state where the locking portion and the second locking portion are engaged. It is characterized by.

  According to a second aspect of the present invention, in the porous filter column according to the first aspect, the first locking portion is formed of a convex portion formed in a ring shape over the entire circumference of the outer periphery of the filter pressing / combining portion. The second locking portion is formed of a concave portion that is formed in a ring shape on the inner peripheral surface of the cylindrical portion of the lower container and engages with the convex portion.

  According to a third aspect of the present invention, in the porous filter column according to the second aspect, the width of the convex portion is 0.2 mm or more and 2 mm or less.

  According to a fourth aspect of the present invention, in the porous filter column according to any one of the first to third aspects, the porous filter has a nucleic acid adsorption ability.

  The invention according to claim 5 is a reagent cartridge in which a reagent for separating and purifying nucleic acid from a subject is contained, and a liquid is dispensed using a dispensing chip, and the subject containing portion that contains the subject 5. A reagent storage section that stores the reagent, a waste liquid storage section that stores a waste liquid generated in the separation and purification, and the porous filter column according to claim 1. And

  The invention of claim 6 is a nucleic acid purification kit comprising the reagent cartridge according to claim 5 and a dispensing chip container for housing a plurality of the dispensing chips.

  According to the porous filter column used in the reagent cartridge and the nucleic acid purification kit of the present invention, the porous filter is securely held in the column by providing the upper container and the lower container with the engaging portions that engage with each other. However, the permeability of the solution and the amount of movement of the fluid are maintained high, and no gaps are generated. Even a soft filter can be assembled without applying heat or using a separate part. Since the fitting depth to the lower container can be stopped at a predetermined position, the porous filter does not crack and can be easily assembled without selecting an operator.

It is a perspective view which shows one Embodiment of the reagent cartridge using the porous filter column concerning this invention. It is a perspective view which shows the reagent cartridge and dispensing tip rack in the nucleic acid purification kit concerning this invention. It is a longitudinal cross-sectional view which decomposes | disassembles and shows the component of the porous filter column concerning this invention. It is a longitudinal cross-sectional view which decomposes | disassembles and shows some components of the porous filter column concerning this invention. FIG. 4 is a longitudinal sectional view showing an intermediate stage of assembly of the porous filter column shown in FIG. 3. FIG. 4 is a longitudinal sectional view showing an end stage of assembly of the porous filter column shown in FIG. 3. It is the longitudinal cross-sectional view which expanded and showed the engagement part of the convex cross-section of the porous filter column of completion | finish of assembly, and the convex latching | locking part of a filter holding | maintenance combined part, and the concave latching part of a cylindrical part. It is a partially cutaway perspective view showing another example of the lower container constituting the porous filter column according to the present invention.

  Hereinafter, an embodiment of the present invention will be described in detail with reference to the drawings. Hereinafter, a nucleic acid purification kit including a reagent cartridge used for separating and purifying nucleic acid and a dispensing chip container, in which the porous filter column according to the present invention is housed, will be described.

  As shown in FIGS. 1 and 2, the nucleic acid purification kit 51 contains a plurality of reagent cartridges 100 containing reagents for extracting nucleic acids from a specimen and a plurality of dispensing chips 201 for dispensing liquids. The dispensed tip rack (dispensed tip container) 200 is provided. In the present embodiment, the dispensing tip rack 200 includes a plurality of dispensing tips 201 having the same diameter and the same size, and the liquid stored in the reagent cartridge 100 is dispensed or stirred by any of the plurality of dispensing tips 201. In operation, the dispensing tip 201 prevents cross contamination between liquids. The dispensing tip rack 200 is a container for collecting the dispensing tips 201 after use. After the dispensing tips 201 are used, the dispensing tips 201 are used as infectious wastes. Can be discarded.

  FIG. 1 shows a perspective view of a reagent cartridge 100. The reagent cartridge 100 has a rectangular shape and a main body 101 formed in a substantially box shape having an open upper surface, and a longitudinal direction of the main body 101. And a claw portion 102 formed to protrude outward on the outer surface of the intermediate portion. The claw portion 102 is for engaging with a part of the nucleic acid analyzer so that the reagent cartridge 100 does not fall over when the reagent cartridge 100 is set in the nucleic acid analyzer, for example.

  A thin film-like sealing film 103 that is removed during use is wound around the upper surface and side surfaces of the main body 101. The upper surface opening of the main body 101 is sealed with a sealing film 103, thereby holding the porous filter column 10 and the like housed in the main body 101 so as not to drop off from the main body 101. The sealing film 103 also has a function of preventing foreign matter such as dust or dirt from entering the main body 101 until use.

  FIG. 2 shows a perspective view of the reagent cartridge 100 and the dispensing tip rack 200 with the sealing film 103 removed, and a sample well (in which a specimen such as a biological sample is put in the main body 101). (Analyte accommodating part) 110, a reagent well part 120 accommodating a reagent for extracting nucleic acid from the object, and a waste well for discarding an unnecessary solution separated in the step of extracting nucleic acid from the object A (waste liquid storage unit) 130 and a recovery well 140 for recovering a nucleic acid extracted from the subject are integrally formed in a state of being arranged in the longitudinal direction of the main body 101. Further, inside the main body 101, a holding unit 160 that accommodates the porous filter column 10 is integrally formed so as to be positioned between the sample well (analyte accommodating unit) 110 and the recovery well 140.

  The holding unit 160 is in an initial position where the porous filter column 10 is accommodated in the reagent cartridge 100. An absorber (not shown) that absorbs liquid can be provided at the bottom of the holding unit 160. The absorbent body comes into contact with the outer surface of the porous filter column 10 on the discharge port (described later) side when the porous filter column 10 is accommodated in the holding unit 160. For this reason, for example, when the cleaning liquid adheres to the outer surface of the discharge port when the cleaning liquid is supplied into the porous filter column 10, the cleaning liquid is absorbed by removing the cleaning liquid by disposing an absorber on the bottom of the storage unit 160. can do.

  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. Yes. 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 are sealed with the well sealing film 104 until use. The well sealing film 104 is preferably configured to suppress gas permeation and be easily broken by piercing the dispensing tip 201. Examples of the material include a metal thin film and a plastic film. Can be used.

  The reagent well 121 accommodates a lysing solution 121A that dissolves a biological material such as a cell membrane. The reagent well 122 accommodates a lysate 122A that dissolves biological substances such as cytoplasm that cannot be completely lysed by the lysate 121A and clog the carrier. In the reagent wells 123 and 124, washing liquids 123A and 124A for washing away unnecessary substances other than the nucleic acid adsorbed on the carrier are accommodated, respectively. The reagent well 125 accommodates an eluent 125A for eluting nucleic acid from the carrier. The reagent well 126 contains a diluent 126A for adjusting the nucleic acid concentration in the eluate.

  The oil well 127 contains, for example, a well-known silicon oil 127A used in a layered manner on a reaction solution in a PCR reaction. As the oil 127A, for example, mineral oil or silicon oil can be preferably used.

  As shown in FIG. 2, the waste liquid well 130 has a recess corresponding to the outer shape of the porous filter column 10. Although the details will be described later, it is possible to support the porous filter column 10 so that it does not fall over even when the porous filter column 10 is inserted into the waste liquid well 130 and the discharge port does not contact the waste liquid in the waste liquid well.

  Similar to the waste well 130, the recovery well 140 has a recess corresponding to the outer shape of the porous filter column 10, and can support the porous filter column 10 inserted into the recovery well 140. The bottom of the recovery well 140 has a storage unit (not shown) for storing the nucleic acid solution eluted from the carrier of the porous filter column 10 by the eluent 125A.

  The waste well 130 and the recovery well 140 are arranged adjacent to each other in the reagent cartridge 100. This is for shortening the moving distance of the porous filter column 10 when the porous filter column 10 is washed in the waste liquid well 130 and then moved to the recovery well 140. As a result, the possibility of contaminating other parts of the reagent cartridge 100 can be reduced.

  Next, the porous filter column 10 of the present invention housed in the reagent cartridge 100 will be described.

  As shown in FIG. 3, the porous filter column 10 is coupled to a cylindrical upper container 11 in which a solution such as a sample liquid or a washing solution is accommodated and a lower part of the upper container 11 so as to be assembled. A funnel-shaped lower container 12 constituting the bottom, a disk-shaped support member 22 disposed in the lower container 12 and positioned on a plane orthogonal to the axial direction of the upper container 11, and a support member 22 in the lower container 12 And a porous filter 21 that is placed on the entire surface on the surface that is overlaid and orthogonal to the axial direction of the upper container 11.

  As shown in FIGS. 3 and 4, the upper container 11 has a length (for example, 5 mm to 50 mm) and an outer diameter (for example, 5 mm to 20 mm) necessary for filtering the liquid and extracting a specific substance in the liquid, and 0. A cylindrical part 1111 having a wall thickness of 5 mm to 3 mm, and a cylindrical filter presser combined part 1112 integrally formed at the lower end of the cylindrical part 1111 extending from the lower end in the axial direction of the cylindrical part 1111. And a gripped member 1113 provided on the outer peripheral surface of the cylindrical portion 1111 so as to cover the entire circumference. The cylindrical portion 1111 has an opening 1114 for dispensing reagents such as a use solution and a cleaning solution and introducing pressurized air to the upper end thereof, and an outer periphery of the upper end outer periphery of the cylindrical portion 1111 over the entire circumference. It has the collar part 1115 which protruded and was formed integrally.

  The gripped member 1113 becomes a gripping part of the porous filter column 10 by an apparatus carrier moving means (not shown) when the porous filter column 10 is moved from the holding part 160 of the reagent cartridge 100 to the recovery well 140, for example. The porous filter column 10 is used to support the porous filter column 10 so that the porous filter column 10 does not fall over when the porous filter column 10 is inserted into the waste liquid well (waste liquid storage unit) 130 of the reagent cartridge 100 or the like. Further, the grasped member 1113 also serves as a reinforcement when the porous filter column 10 is grasped by the apparatus carrier moving means.

The filter holding / combining portion 1112 has a function of holding the porous filter 21 including the support member 22 in the porous filter column 10 with a constant pressure by pressing the peripheral edge of the porous filter 21 from above, and the upper part. The container 11 and the lower container 12 have a function of being airtight and liquid tightly coupled.
For this purpose, the filter holding / combining portion 1112 has a length (for example, 5 mm to 20 mm) necessary for pressing and holding the porous filter 21 including the support member 22 and the lower container 12, and an inner diameter of the cylindrical portion 1111. The outer diameter is smaller than the outer diameter of the cylindrical portion 1111 and has the same outer diameter as that of the disk-shaped porous filter 21 and the support member 22 (for example, 5 mm to 20 mm). Further, on the outer peripheral surface of the filter holding / combining portion 1112, the fitting depth of the filter holding / combining portion 1112 to a cylindrical portion 1211 of the lower container 12 to be described later is defined, and the pressing to the peripheral portion of the porous filter 21 is performed. A convex locking portion 1116 (corresponding to the first locking portion described in the claims) for holding the pressure constant is formed in a ring shape over the entire circumference.

The width of the convex locking portion 1116 is preferably 0.2 mm or more and 2 mm or less.
Here, when the width of the convex locking portion 1116 is set in the range of 0.2 mm or more and 2 mm or less, stronger sealing is obtained. Moreover, when the width | variety of the convex latching | locking part 1116 is less than 0.2 mm or exceeds 2 mm, the malfunction that the force required for a fitting will not stop more than needed or a regular position will arise.

  The lower container 12 is integrally formed at the lower end of the cylindrical portion 1211 and the cylindrical portion 1211 that is airtightly and liquid-tightly fitted to the outer periphery of the filter holding / combining portion 1112, and from the lower end of the cylindrical portion 1211 to the cylindrical portion 1211. A bottom surface portion 1212 that inclines in a descending gradient from the vicinity of the lower end of the cylindrical portion 1211 in the axial direction toward the axial center of the cylindrical portion 1211, and a central portion that coincides with the axial center of the cylindrical portion 1211 of the bottom surface portion 1212. And a discharge port 1213.

  The cylindrical portion 1211 of the lower container 12 has the same outer diameter as the outer diameter of the cylindrical portion 1111 of the upper container 11 and the same inner diameter as the outer diameter of the filter pressing / coupling portion 1112. Further, on the inner peripheral surface of the cylindrical portion 1211, a concave locking portion 1214 that engages with the convex locking portion 1116 of the filter presser combined portion 1112 (in the second locking portion described in the claims). Equivalent) is formed in a ring shape over the entire circumference. Furthermore, a ring-shaped carrier portion 1215 that engages with the outer peripheral edge portion 22a on the lower surface side of the support member 22 is formed over the entire circumference at the inner corner portion of the joint portion between the cylindrical portion 1211 and the bottom surface portion 1212. .

  The reason for providing the engaging part for engaging the filter presser combined part 1112 and the cylindrical part 1211 of the lower container 12 is that the filter presser combined part 1112 is fitted to the cylindrical part 1211 of the lower container 12. Then, when the convex locking portion 1116 and the concave locking portion 1214 are engaged with each other, the fitting depth of the filter presser combined portion 1112 to the cylindrical portion 1211 is defined, and the filter presser combined connection The lower end of the portion 1112 is held in contact with the peripheral edge of the porous filter 21 from above so that a constant pressing force is applied. This is to enable assembly of the porous filter column 10 by fitting the filter holding / combining portion 1112 and the cylindrical portion 1211 of the lower container 12 without causing deformation or cracking of the porous filter 21. .

  The material for forming the upper container 11 and the lower container 12 is not limited as long as it does not dissolve in the solvent used for the sample solution and does not affect the sample or reagent in the solution, but in particular, polypropylene, polycarbonate, If a resin material containing any of acryl is used, good visible light permeability can be secured, and the state of the solution can be confirmed. As polypropylene, homopolypropylene or a random copolymer of polypropylene and polyethylene can be used. Moreover, as an acryl, the copolymer of monomers, such as polymethyl methacrylate or methyl methacrylate, and other methacrylic acid ester, acrylic acid ester, styrene, can be used. Moreover, when using these resin materials, the heat resistance and intensity | strength of a chip | tip can also be ensured.

  The support member 22 prevents the porous filter 21 from being deformed in the cylindrical portion 1211 of the lower container 12. As the porous filter 21, a porous material having a large surface area is used because the sample solution penetrates into the inside by using a material having a hydrophilic group on the surface so that the biological sample is chemically adsorbed. Those formed in the film form are preferred. Also, the nucleic acid is adsorbed and held during washing with the washing liquid, and the nucleic acid adsorption force is weakened and separated during the collection with the collecting liquid. As the porous material in the present invention, a material in which fibrous materials such as glass wool are superimposed may be used.

The shape of the porous filter 21 may be any shape that does not cause a gap between the porous filter 21 and the cylindrical portion 1211 when installed horizontally in the cylindrical portion 1211 of the lower container 12. In the present embodiment, the porous filter 21 has a circular disk shape having an outer diameter equal to the inner diameter of the cylindrical portion 1211 in plan view. Further, the film thickness of the porous filter 21 varies depending on the type of hydrophilic group, the surface area of the porous material, the type of sample to be adsorbed, and the like. It is sufficient to set a value that can adsorb as many samples as necessary.
Moreover, the porous filter 21 may be one sheet or a plurality of sheets. Furthermore, when installing two or more sheets, the material may be the same or different.

  The material of the porous filter 21 is not particularly limited as long as it can adsorb a biological substance such as a nucleic acid in the presence of an organic substance. It is preferable to use a material having a hydrophilic group introduced therein. Examples of the inorganic material having a hydrophilic group include silica, a silica derivative obtained by introducing a hydrophilic group into silica, diatomaceous earth, and alumina. Examples of the organic material having a hydrophilic group include polyhydroxyethyl acrylic acid, polyhydroxyethyl methacrylic acid, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, polymethacrylic acid, polyoxyethylene, acetylcellulose, and acetyl having different acetyl values. A mixture of cellulose, an organic material having a polysaccharide structure, or the like can be used.

  Furthermore, the surface of a material having no hydrophilic group such as glass or ceramic may be coated with a material having a hydrophilic group. Examples of the material used for the coating include polyhydroxyethylacrylic acid and polyhydroxyethylmethacrylic. Polymers of organic materials such as acids and salts thereof, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, polymethacrylic acid and salts thereof, polyoxyethylene, acetyl cellulose, and mixtures of acetyl cellulose having different acetyl values are preferred.

  In the present specification, the “hydrophilic group” refers to a polar group capable of interacting with water, and applies to all groups involved in the adsorption of biological substances such as nucleic acids. Such a hydrophilic group may be any group capable of adsorbing a nucleic acid with a polar group having an interaction with water, such as a hydroxyl group, a carboxyl group, a cyano group, an oxyethylene group, an amino group, or a hydrophilic group. Examples include groups in which these hydrophilic groups are modified for the purpose of controlling the above.

  Since the porous filter 21 is pressurized by a pressurizing means at the time of washing / filtering or the like, when the low-strength porous filter 21 is used, the porous filter 21 is bent, and the inner surface of the cylindrical portion 1211 and the porous filter 21 are porous. A gap through which the solution passes may be formed between the filter 21 and the solution may leak from this portion. However, the porous filter 21 may be bent by arranging the support member 22 having high rigidity. Even when the low-strength porous filter 21 is used, the pressure can be suitably applied. Therefore, it is preferable that the support member 22 be 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 material used for the support member 22 is preferably, for example, a filter-like material that is made by sintering resin particles and that allows liquid to pass through, but is not limited thereto, and is not limited to cleaning. It does not dissolve in the solvent used in the step, the rigidity is not lowered by the solvent, a substance that affects the sample, the reagent, or the like does not elute, and the target solution, impurities, or the like can pass therethrough. The support member 22 can be made of the same resin material as that of the upper container 11 and the lower container 12, but is particularly limited as long as it is formed to be porous so that the liquid that has passed through the filter can pass smoothly. Is not to be done.

Next, a method for assembling the porous filter column 10 will be described with reference to FIGS.
First, as shown in FIG. 5, the porous filter 21 and the support member 22 are inserted into the cylindrical portion 1211 of the lower container 12 until the outer peripheral edge portion 22 a of the support member 22 is locked to the support portion 1215. Thereafter, as shown in FIG. 6, the filter retainer combined portion 1112 of the upper container 11 is inserted into the cylindrical portion 1211 of the lower container 12 from the lower end side, and the entire upper container 11 is pressed from above to project The coupling portion 1112 serving as a filter presser is pushed into the cylindrical portion 1211 until the engagement portion 1116 is engaged with the concave locking portion 1214, and the coupling portion 1112 serving as a filter presser is fitted into the cylindrical portion 1211.

Here, the engaging part 1116 of the filter press / combination connecting part 1112 and the engaging part 1214 of the lower container 12 are engaged with each other as shown in FIG. In addition to being fitted at a prescribed fitting depth position, the amount of fitting and pushing into the cylindrical portion 1211 of the filter pressing / coupling portion 1112 is limited. In other words, the fitting amount between the filter holding / combining portion 1112 and the cylindrical portion 1211 is set at a predetermined position by the convex locking portion 1116 and the concave locking portion 1214. As a result, the pressing force applied to the porous filter 21 caused by the lower end of the filter holding / combining portion 1112 coming into contact with the peripheral edge of the porous filter 21 from above causes the porous filter 21 to be deformed or cracked. It is possible to assemble a porous filter column easily and reliably by anyone, regardless of the operator's technical ability and the material of the porous filter. Become.
Further, the engaging portion 1116 of the filter retainer combined portion 1112 and the engaging portion 1214 of the lower container 12 are engaged with each other, whereby the filter retainer combined portion 1112 and the cylindrical portion 1211 are deformed into the porous filter 21. It is possible to feed back to the operator as a touch that it has been fitted in a state that does not cause cracks.

  When the porous filter column 10 assembled as described above is used, first, reagents such as a use solution and a cleaning solution are dispensed from the opening 1114 of the upper container 11, and then pressurized air is supplied to the opening 1114. Introduce from. Thereby, the reagents are adsorbed on the porous filter 21 or passed through or filtered through the porous filter 21. The solution passed through or filtered through the porous filter 21 is discharged from the discharge port 1213 or collected in a separate container through the bottom surface portion 1212 inclined like a funnel of the lower container 12.

Next, the lower container 12 shown in FIG. 8 will be described.
In FIG. 8, the inner surface of the bottom surface portion 1212 constituting the lower container 12 has a radius along the inclined bottom surface portion 1212 from a ring-shaped carrier portion 1215 formed at a joint corner portion between the cylindrical portion 1211 and the bottom surface portion 1212. A plurality of support pieces 1216 having a triangular flat plate shape extending in the direction are integrally formed with a certain interval in the circumferential direction. Each of the carrier pieces 1216 is for supporting the porous filter 21 at a plurality of locations from the lower surface side via the support member 22 and preventing these deformations.

  Regarding the operation when the nucleic acid purification kit 51 including the reagent cartridge 100 and the dispensing tip rack 200 having the porous filter column 10 having the above-described configuration is used, the operation of the porous filter column 10 will be described by taking the separation and purification of nucleic acid as an example. The explanation will be focused on.

  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.

  Subsequently, the 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 from the reagent wells 121 to 126 into the dispensing tip 201, the sealing film 104 sealing the reagent wells 121 to 126 is pierced at the tip of the dispensing tip 201. Then, a through-hole is formed in the sealing film 104 so that various reagents in the reagent wells 121 to 126 can be sucked by the dispensing tip 201.

Since it is necessary to collect the waste liquid, the porous filter column 10 is conveyed to the waste liquid well 130, and the solution in which the cells are lysed is supplied from the sample well 110 to the porous filter column 10.
Since the inner diameter of the recess formed in the waste liquid well 130 is larger than the outer diameter of the entire cylindrical portion 1111 including the gripped member 1113 constituting the upper container 11, the discharge port 1213 and the gripped member 1113 of the porous filter column 10 are Although it is inserted into the well, the gripped member 1113 cannot enter the waste liquid well 130. Therefore, the portion where the gripped member 1113 is provided does not enter the waste liquid well 130, and the discharge port 1213 is the waste liquid in the waste liquid well. It is supported at a height not touching.

  The porous filter column 10 can increase the speed at which the liquid passes through the porous filter 21 by sending gas from the opening 1114 and pressurizing the inside of the column. Then, the solution in which the cells are dissolved passes through the porous filter 21, and the nucleic acid is adsorbed on the porous filter 21. Thereafter, the porous filter 21 is washed with a dissolving solution 122A that dissolves a biological substance such as cytoplasm that cannot be completely dissolved in the dissolving solution 121A and clogs the carrier.

  Next, the cleaning liquids 123A and 124A are supplied to the porous filter 21, and the porous filter 21 is cleaned with the cleaning liquids 123A and 124A. Thereafter, the porous filter column 10 is transported to the recovery well 140, and the eluent 125 A is supplied to the porous filter 21. Thereby, the nucleic acid adsorbed by the porous filter 21 is eluted in the eluent 125A, and the nucleic acid solution containing the nucleic acid is recovered in the recovery well 140.

  Thereafter, when the diluent 126A and the eluate 125A from which the recovered nucleic acid is recovered are mixed, the sample preparation is completed. Thus, the separation and purification of the nucleic acid by the nucleic acid purification kit 51 including the porous filter column 10 is completed.

  In the above-described embodiment, the first locking portion 1116 is constituted by a convex portion formed in a ring shape over the entire circumference of the outer periphery of the filter press / joining portion 1112, and the second locking portion 1214 is formed in the lower container 12. Although the case where it comprised from the recessed part formed in the ring shape over the perimeter on the inner peripheral surface of the cylindrical part 1211 of this invention was demonstrated, this invention is not limited to this, For example, a 1st latching | locking part is a ring contrary to the above. The second locking portion may be composed of a ring-shaped convex portion, and various modifications may be made without departing from the requirements described in the claims. Is configured such that the lower end of the filter pressing and combining portion is in contact with the upper surface of the peripheral edge of the porous filter with a constant pressing force when the first locking portion and the second locking portion are engaged. Anything is acceptable.

DESCRIPTION OF SYMBOLS 10 ... Porous filter column 11 ... Upper container 1111 ... Cylindrical body 1112 ... Combined part 1113 ... Filter holding member 1114 ... Opening part 1115 ... Eaves part 1116 ... Convex-shaped locking part 12 ... Lower container 1211 ... Cylindrical part DESCRIPTION OF SYMBOLS 1212 ... Bottom part 1213 ... Ejection port 1214 ... Recessed latching part 1215 ... Supporting part 1216 ... Rib-like support piece 21 ... Porous filter 22 ... Supporting member 51 ... Nucleic acid purification kit 100 ... Reagent cartridge 110 ... Sample well (covered) Specimen storage unit)
121, 122, 123, 124, 125, 126 ... Reagent well (reagent storage unit)
130 ... Waste liquid well (waste liquid container)
200: Dispensing tip rack 201: Dispensing tip

Claims (6)

A cylindrical upper container;
A lower container coupled to the lower part of the upper container;
A support member disposed in the lower container;
A porous filter that is supported by the support member in the lower container and is disposed in the entire region on a surface orthogonal to the axial direction of the upper container;
The upper container has a cylindrical portion having an opening at the upper end, and a filter press combined joint portion having a cylindrical shape provided at the lower outer peripheral surface of the cylindrical portion,
The lower container has a cylindrical portion that is tightly fitted to the outer periphery of the filter press combined joint portion, and a bottom portion that connects the lower portion of the cylindrical portion and has a discharge port formed.
A first locking portion is formed on the outer peripheral surface of the filter pressing and combining portion,
On the inner peripheral surface of the cylindrical portion of the lower container, there is a second locking portion that engages with the first locking portion and defines the fitting depth of the filter pressing and coupling portion to the cylindrical portion of the lower container. Formed,
In a state in which the filter press combined part is fitted to the cylindrical part of the lower container and the first locking part and the second locking part are engaged, the lower end of the filter press combined part is the porous It is configured to abut on the upper surface of the peripheral edge of the quality filter with a constant pressing force,
A porous filter column characterized by that.   The first locking portion comprises a convex portion formed in a ring shape over the entire circumference of the outer periphery of the filter-holding combined portion, and the second locking portion is formed on the inner circumferential surface of the cylindrical portion of the lower container. 2. The porous filter column according to claim 1, wherein the porous filter column is formed in a ring shape over the entire circumference and is formed of a concave portion that engages with the convex portion.   The porous filter column according to claim 2, wherein a width of the convex portion is 0.2 mm or more and 2 mm or less.   The porous filter column according to claim 1, wherein the porous filter has a nucleic acid adsorption ability. A reagent cartridge that contains a reagent for separating and purifying nucleic acid from a specimen and dispenses a liquid using a dispensing chip,
A subject storage section for storing the subject;
A reagent storage section for storing the reagent;
A waste liquid storage unit for storing a waste liquid generated in the separation and purification;
The porous filter column according to any one of claims 1 to 4,
A reagent cartridge comprising: A reagent cartridge according to claim 5;
A dispensing tip container for housing a plurality of the dispensing tips;
A nucleic acid purification kit comprising:

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