Container, culture kit, nucleic acid analysis system, and cell culture method

Abstract

To provide a container, a culture kit, and a nucleic acid analysis system which can suppress contamination risk or operation mistake without being influenced by an operator's skill.SOLUTION: A container 10 comprises: a container body 20 having an opening 20a; a lid 30 freely detachably attached to the container body 20, and sealing the opening 20a; a shaft body 40 provided on the lid 30, and extending into the container body 20; and a filter fixing part 50 provided on the shaft body 40, and hooking a collecting filter to be fixed to the shaft body 40.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a container, a culture kit, and a nucleic acid analysis system. and cell culture method This is regarding. [Background technology]

[0002] The following Patent Document 1 discloses a nucleic acid extraction method in which a membrane filter is used to collect a sample containing nucleic acid and extract the nucleic acid from the collected sample. The membrane filter is set in a collection section including a funnel and a filtration bottle, and collects the sample by filtering a liquid containing the sample poured into the collection section. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2019 / 043779 Summary of the Invention [Problem to be solved by the invention]

[0004] After collecting the sample, the membrane filter is retrieved with tweezers and transferred to a Petri dish for cell culture. However, when picking up the membrane filter with tweezers, depending on the operator's skill, the membrane filter may be accidentally dropped onto the floor. In addition, if the tweezers are used multiple times, the sample attached to the tweezers may adhere to other membrane filters, causing contamination.

[0005] The present invention has been made in consideration of the above circumstances, and provides a container, a culture kit, and a nucleic acid analysis system that are not dependent on the skill of an operator and can reduce the risk of contamination and operational errors. and cell culture method The purpose is to provide. [Means for solving the problem]

[0006] (1) A container according to one embodiment of the present invention comprises a container body having an opening, a lid body that is removably attached to the container body and seals the opening, a shaft body that is attached to the lid body and extends toward the inside of the container body, and a filter fixing portion that is attached to the shaft body and hooks a filter and fixes it to the shaft body.

[0007] (2) In the container described in (1) above, the filter fixing portion may have a tip extending in a direction other than the axial direction along which the shaft body extends.

[0008] (3) In the container described in (1) above, the filter fixing portion may have a slit piece having a slit formed therein.

[0009] (4) The container according to any one of (1) to (3) above may further include a second filter fixing part provided at a position different from the filter fixing part in the circumferential direction of the shaft.

[0010] (5) In the container described in (4) above, the second filter fixing portion may have a tip extending in a direction other than the axial direction along which the shaft body extends.

[0011] (6) In the container described in (1) above, the pointed portion may have a protrusion extending in a direction other than the direction in which the pointed portion extends.

[0012] (7) A culture kit according to one aspect of the present invention includes a filter for collecting cells, and a container according to any one of (1) to (6) above in which the filter and a culture medium are enclosed.

[0013] (8) A nucleic acid analysis system according to one embodiment of the present invention collects cells using a container described in any one of (1) to (6) above, encloses a culture medium in the container, extracts nucleic acid from the cultured cells, and analyzes the extracted nucleic acid.

[0014] (9) A nucleic acid analysis system according to one embodiment of the present invention extracts nucleic acid from cells collected and cultured using the culture kit described in (7) above, and analyzes the extracted nucleic acid. Effect of the Invention

[0015] According to one aspect of the present invention, it is possible to obtain a container, a culture kit, and a nucleic acid analysis system that are not dependent on the skill of an operator and that can reduce the risk of contamination and operational errors. [Brief description of the drawings]

[0016] [Figure 1] 1 is a schematic diagram of a nucleic acid analysis system according to a first embodiment. [Diagram 2] FIG. 2 is a vertical cross-sectional view of a container used in the bacteria collection system according to the first embodiment. [Diagram 3] 3 is a cross-sectional view taken along line III-III in FIG. 2. [Figure 4] FIG. 2 is a process diagram from sample collection to culture according to the first embodiment. [Diagram 5] FIG. 1 is a vertical sectional view of a bacteria collection system according to a first embodiment. [Figure 6] FIG. 4 is an explanatory diagram showing how a collection filter is collected from the bacteria collection system according to the first embodiment. [Figure 7] 3 is a vertical cross-sectional view of a container in which a collection filter according to the first embodiment has been collected. FIG. [Figure 8] 8 is a cross-sectional view taken along line VIII-VIII in FIG. 7. [Figure 9] FIG. 11 is a vertical cross-sectional view of a container according to a second embodiment. [Figure 10] FIG. 11 is a vertical cross-sectional view of a shaft body according to a second embodiment. [Figure 11] FIG. 11 is an explanatory diagram showing how the collection filter is collected from the bacteria collection system according to the second embodiment. [Figure 12] FIG. 11(b) is a view taken along line A in FIG. [Figure 13] FIG. 11 is a vertical cross-sectional view of a container according to a third embodiment. [Figure 14]14 is a cross-sectional view taken along line XIV-XIV in FIG. 13. [Figure 15] FIG. 4 is a cross-sectional view of a container according to a modified example of the first embodiment. [Figure 16] FIG. 4 is a cross-sectional view of a container according to a modified example of the first embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Hereinafter, a container, a culture kit, and a nucleic acid analysis system according to embodiments of the present invention will be described in detail with reference to the drawings. First, an overview of the embodiments of the present invention will be described, and then the details of the embodiments of the present invention will be described.

[0018] 〔overview〕 The nucleic acid extraction method described in the above-mentioned Patent Document 1 collects a sample containing nucleic acid by a membrane filter method. This membrane filter method uses a filter funnel and a suction system, passes a liquid through a membrane filter set in the filter funnel, and collects a sample such as microorganisms contained in the liquid on the surface of the membrane filter.

[0019] Next, the membrane filter with the sample collected is transferred to a Petri dish using tweezers, and placed on an absorbent pad soaked in liquid medium in the Petri dish.The sample is then grown on the surface of the membrane filter by the nutrients absorbed from the absorbent pad through the membrane filter.

[0020] However, when collecting a membrane filter with tweezers, there are the following problems. For example, when picking up the membrane filter with the tweezers, the membrane filter may be dropped by mistake. In addition, when the tweezers are used multiple times, the sample attached to the tweezers may adhere to other membrane filters, causing contamination. In addition, the task of picking up the membrane filter with the tweezers is itself cumbersome.

[0021] Furthermore, culturing samples in Petri dishes poses the following problems. For example, when culturing liquid in a Petri dish, an absorbent pad is required to prevent leakage of the culture medium. In addition, when culturing on an absorbent pad, the membrane filter absorbs the liquid culture medium, so nutrients are only supplied to the sample little by little, and it takes time for the sample to grow. As described below, when a liquid sample is analyzed for the presence of fungi (bacteria or fungi) using a liquid analysis system, it is desirable to analyze a sample collected by a membrane filter or a liquid containing a culture obtained by culturing and growing the sample as a sample. However, when an absorbent pad or the like is used to prevent leakage of the culture medium as described above, it is difficult to recover the sample contained in the culture medium absorbed on the membrane filter surface or the absorbent pad, or the culture in a liquid state. Therefore, a container that holds a membrane filter in a culture medium so that the culture of the collected sample can be cultured in the liquid without leakage of the culture medium, or a culture kit using the same, has been desired.

[0022] In the embodiment of the present invention, in a container, a culture kit, and a nucleic acid analysis system, a filter fixing part is provided on a shaft body, and a collection filter with cells attached thereto is hooked onto the shaft body and wound around the shaft body for recovery. Then, the shaft body with the collection filter wound around it is placed in a container containing a culture medium for culture. This makes it possible to reduce the risk of contamination and operational errors without being dependent on the skill of the operator. Also, efficient culture is possible in a liquid culture medium using the collection filter. Furthermore, efficient culture is possible by shaking the liquid culture medium containing the collection filter.

[0023] [First embodiment] FIG. 1 is a schematic diagram of a nucleic acid analysis system 1 according to a first embodiment. As shown in FIG. 1, a nucleic acid analysis system 1 includes a bacteria recovery system 2, a nucleic acid extraction system 3, a hybridization reaction system 4, and a detection system 5.

[0024] The microorganism collection system 2 is a system that collects microorganisms (bacteria, fungi, etc.) contained in the sample 100 from the sample 100. For example, in the case of testing a beverage, the sample 100 is the produced beverage, the water for producing the beverage, or a liquid in the process of producing the beverage. Alternatively, the sample 100 may be a liquid from which microorganisms have been collected from a cotton swab or the like used to wipe the test environment in order to test the presence or absence of bacterial contamination in the production environment and the level of contamination.

[0025] The bacteria can be collected, for example, by applying pressure or vacuum to the collected liquid and filtering it with a collection filter, which will be described later. When collecting bacteria or fungi, the collection filter may be a membrane filter with a pore size of 0.22 μm to 0.45 μm. After collecting the bacteria with the collection filter, the collection filter may be immersed in a culture solution in which the bacteria are cultured, and the culture solution in which the bacteria are cultured may be used as the sample 100 for the next step (nucleic acid extraction system 3). Alternatively, a liquid in which the bacteria are collected by centrifugation or the like, or a liquid in which aggregates collected by centrifugation or the like are dissolved, may be used as the sample 100 for the next step. Alternatively, the liquid containing the collection filter may be vibrated, and the liquid in which the bacteria are suspended may be used as the sample 100 for the next step.

[0026] The nucleic acid extraction system 3 is a system that breaks (dissolves) the membrane structure of cells in the sample 100 and extracts the nucleic acid of the cells. The sample 100 from which the nucleic acid has been extracted may be mixed with a liquid containing other nucleic acids that react with the extracted nucleic acid. The other nucleic acids may be nucleic acids to which a site that exhibits fluorescence, luminescence, or quenching action under specific conditions has been added in order to detect them in the detection step (detection system 5) described below. These may be mixed with the sample 100 before processing with the nucleic acid extraction system 3, or may be mixed with the sample 100 after processing with the nucleic acid extraction system 3.

[0027] The hybridization reaction system 4 is a system that causes a hybridization reaction of the nucleic acid in the sample 100. In this step, the sample 100 is heated to, for example, 60° C. and stirred to cause a hybridization reaction that matches with the other nucleic acid described above. In this reaction, for example, a site that has a fluorescent, luminescent, or quenching effect under specific conditions, which is given to the other nucleic acid described above, reacts with the nucleic acid in the sample 100, thereby causing the fluorescence, luminescence, or quenching effect to be expressed.

[0028] In addition, by designing the structure of the other nucleic acid described above to react with a specific nucleic acid, it can be made to react only with nucleic acid contained in, for example, a specific bacterium, fungus, or the like in the sample 100. In other words, in the processing of the hybridization reaction system 4, by using another nucleic acid that reacts with the specific nucleic acid, it is possible to make the fluorescence, luminescence, or quenching effect imparted to the other nucleic acid manifest only when the sample 100 contains a specific bacterium, fungus, or the like.

[0029] The detection system 5 detects the presence or absence, and the degree of, fluorescence, luminescence, or quenching action exhibited in the sample 100 treated by the hybridization reaction system 4. The detection system 5, for example, excites the fluorescence action exhibited in the nucleic acid of the sample 100 with an excitation laser light, and detects the excited fluorescence with a high-sensitivity camera.

[0030] Alternatively, the detection system 5 detects the luminescence action expressed by the nucleic acid of the sample 100 with a high-sensitivity camera. Alternatively, the detection system 5 detects the degree to which the fluorescence or luminescence imparted to the vicinity of the site to which the quenching action is imparted is quenched with a high-sensitivity camera, which is the quenching action expressed by the nucleic acid of the sample 100. Regarding this detection method, for example, a method such as that described in JP 2020-74726 A may be adopted.

[0031] The nucleic acid analysis system 1 uses a series of systems as described above to determine whether a particular microorganism (bacteria, fungi, etc.) is present in the sample 100 or to analyze its concentration.

[0032] Fig. 2 is a vertical cross-sectional view of the container 10 used in the bacteria collection system 2 according to the first embodiment. Fig. 3 is a cross-sectional view taken along line III-III shown in Fig. 2. As shown in these figures, the container 10 comprises a container body 20, a lid body 30, a shaft body 40, and a filter fixing part 50.

[0033] As shown in FIG. 2, the container body 20 is formed into a cylindrical shape with a bottom. The lid body 30 is formed into a cylindrical shape with a top. The central axes O of the container body 20 and the lid body 30 coincide with each other. The shaft body 40 extends along the central axis O. In the following description, the direction in which the central axis O extends is referred to as the axial direction. In a plan view seen from the axial direction, a direction intersecting the central axis O is referred to as the radial direction. In addition, a direction rotating around the central axis O is referred to as the circumferential direction.

[0034] The container body 20 has an opening 20a at the upper end. The container body 20 includes a body 21 and a bottom 22. The body 21 extends in the axial direction with a constant inner diameter and outer diameter. The bottom 22 is formed in a hemispherical shape and is connected to the lower end of the body 21. The bottom 22 may be formed in an inverted cone shape, for example. The tip of the inverted cone may be curved, such as a hemispherical shape. The body 21 may have an inclined shape in which the inner and outer diameters become slightly smaller toward the lower end, rather than a constant outer diameter.

[0035] The lid 30 is detachably attached to the container body 20 and seals the opening 20a. The lid 30 includes a top wall 31 and a peripheral wall 32. The peripheral wall 32 is formed in a cylindrical shape and detachably engages with the outer peripheral surface of the upper end of the container body 20. In addition, when a male screw (not shown) is formed on the outer peripheral surface of the upper end of the container body 20, for example, a female screw (not shown) that screws into the male screw may be formed on the inner wall surface of the peripheral wall 32. Alternatively, the peripheral wall 32 of the lid 30 may be configured to enter the inside of the opening 20a of the container body 20, as opposed to the illustrated configuration. In this case, a female screw (not shown) may be formed on the inner peripheral surface of the upper end of the container body 20, and a male screw (not shown) that screws into the female screw may be formed on the outer wall surface of the peripheral wall 32.

[0036] The top wall 31 is formed in a disk shape and is connected to the upper end of the peripheral wall 32 to close the upper end opening of the peripheral wall 32. A fitting hole 33 into which the shaft body 40 fits is formed in the lower surface 31a of the top wall 31 (the inner surface of the lid body 30). The fitting hole 33 is formed in the center position in the radial direction of the top wall 31. Note that the lid body 30 and the shaft body 40 do not have to be separate parts combined by fitting them together, and may be one piece.

[0037] The shaft body 40 is provided on the lid body 30 and extends linearly toward the inside of the container body 20. The upper end 41 of the shaft body 40 fits into the fitting hole 33 of the lid body 30. On the other hand, the lower end 42 of the shaft body 40 does not contact the bottom 22 of the container body 20, but extends to the vicinity of the bottom 22. The shaft body 40 is, for example, a solid shaft. The shaft body 40 is formed in a cylindrical shape with a constant outer diameter. Alternatively, the shaft body 40 may be formed in a rod shape other than a cylindrical shape.

[0038] The filter fixing part 50 is provided on the shaft body 40. The filter fixing part 50 has a tip 51 that hooks and fixes a collection filter 101 (see Figs. 6 and 7) described below around the periphery of the shaft body 40. The tip 51 is provided to protrude from the peripheral surface 40a of the shaft body 40, as shown in Fig. 3.

[0039] The tip 51 extends in a radial direction intersecting the central axis O at a right angle. The tip 51 is formed in a conical shape that tapers toward the radially outward direction. The tip 51 may be formed in a needle shape with only the tip being sharp. The tip 51 may extend in a direction other than the axial direction in which the shaft body 40 extends, for example, in an oblique direction intersecting the central axis O at an acute angle or an obtuse angle.

[0040] The apex 51 further has a protrusion 52 extending in a direction other than the direction in which the apex 51 extends. The protrusion 52 is formed in a plate shape extending from approximately the middle of the inclined surface of the conical apex 51 in a direction perpendicular to the direction in which the apex 51 extends. The protrusion 52 may be an elbow shape bent in an L-shape with respect to the apex 51, or a hook shape like a fish hook, as long as it is in a direction other than the direction in which the apex 51 extends (radial direction). The protrusion 52 shown in FIG. 3 is a plate shape having a constant thickness in the radial direction. As shown in FIG. 2, the protrusion 52 is formed in a disk shape when viewed from the direction in which the apex 51 extends.

[0041] In the first embodiment, the shaft body 40, the apex 51, and the protrusion 52 are integrally molded products, but they may be assembled by fitting separate parts. For example, the protrusion 52 may be a ring member that fits skewer-like into the apex 51. The apex 51 may be a cone member implanted on the peripheral surface 40a of the shaft body 40, or a plate member such as a triangular plate, or a pyramidal member such as a triangular pyramid or a tetrahedron.

[0042] 3, a second filter fixing part 60 is provided on the peripheral surface 40a of the shaft body 40 at a position different from the filter fixing part 50 in the circumferential direction. The second filter fixing part 60 has a peak 61 and a protrusion 62, and has a configuration similar to that of the above-mentioned filter fixing part 50. In a plan view seen from the axial direction, the second filter fixing part 60 has a positional relationship that is point-symmetrical with the filter fixing part 50 about the central axis O. Note that the second filter fixing part 60 is not limited to this positional relationship as long as it is disposed in a different position from the filter fixing part 50 in the circumferential direction.

[0043] Next, a method for recovering the collection filter 101 and a method for culturing cells using the above-mentioned container 10 will be described. Fig. 4 is a process diagram from collection to culture of a sample 100 according to the first embodiment. Fig. 5 is a vertical cross-sectional view of the bacteria collection system 2 according to the first embodiment. Fig. 6 is an explanatory diagram showing a state in which a collection filter 101 is collected from the bacteria collection system 2 according to the first embodiment.

[0044] First, the configuration of the bacteria collection system 2 will be described with reference to Fig. 5. As shown in Fig. 5, the bacteria collection system 2 includes a lower filter part 70 and an upper filter part 80 that sandwich a collection filter 101 from above and below. The collection filter 101 is formed in a circular shape in a plan view. The lower filter part 70 and the upper filter part 80 are connected via a connecting part 90 in the axial direction along a central axis O1 that passes through the center of the collection filter 101 as a common axis.

[0045] The filter lower part 70 includes a cylindrical filtrate collection container 71 with a bottom, and a base plate 72 attached to the upper opening of the filtrate collection container 71. The filtrate collection container 71 and the base plate 72 may be integrated. A suction port 71a is provided on the side wall of the filtrate collection container 71. A reduced pressure suction tube 73 is connected to the suction port 71a. A suction device main body such as a vacuum pump (not shown) is connected to the reduced pressure suction tube 73.

[0046] The base plate 72 is formed with a plurality of through holes 72a that communicate with the inside of the filtrate recovery container 71. An expanded diameter portion 72b is formed in some of the plurality of through holes 72a. The expanded diameter portion 72b is a groove with a larger diameter than the through hole 72a. The expanded diameter portion 72b has a diameter and depth that allows the above-mentioned peaks 51, 61 and protrusions 52, 62 of the filter fixing part 50 and the second filter fixing part 60 to be inserted therein. The expanded diameter portion 72b may be formed on the upper surface of the base plate 72 as an independent groove that does not communicate with the through hole 72a.

[0047] A filter positioning portion 72c that surrounds the multiple through holes 72a and the enlarged diameter portion 72b is formed on the upper surface of the base plate 72. The filter positioning portion 72c shown in Fig. 5 is provided in an annular shape centered on the central axis O1. The filter positioning portion 72c has an inner diameter that is slightly larger than the outer diameter of the collection filter 101.

[0048] The filter positioning portion 72c is an annular protrusion provided on the upper surface of the base plate 72 with a height equal to or greater than the thickness of the collection filter 101. The filter positioning portion 72c may be formed as a ring in the circumferential direction about the central axis O1, or may not be formed as a ring but may be a group of multiple protrusions provided at intervals in the circumferential direction. The filter positioning portion 72c may be a groove that is recessed relatively to the upper surface of the base plate 72.

[0049] The filter upper portion 80 includes a support plate 81 having a through hole 81a formed therein penetrating in the axial direction along which the central axis O1 extends, and a cylindrical sample injection portion 82 attached to the support plate 81 and having an interior communicating with the through hole 81a. The through holes 81a of the support plate 81 are disposed opposite each of the multiple through holes 72a of the base plate 72 with the collection filter 101 interposed therebetween.

[0050] A sealing portion is disposed on the lower surface of the support plate 81, which presses the outer periphery of the collection filter 101 against the upper surface of the base plate 72. The sealing portion may be, for example, an annular convex portion disposed on the support plate 81, or a tubular convex portion having a lower elastic modulus than the support plate 81. Alternatively, the sealing portion may be an O-ring 83 disposed in a groove portion on the lower surface of the support plate 81, as in this embodiment. The O-ring 83 is disposed radially inside the filter positioning portion 72c of the base plate 72. The O-ring 83 is sandwiched between the base plate 72 and the support plate 81, and seals the gap so that the sample 100 does not leak from the connecting portion between the filter lower portion 70 and the filter upper portion 80. The O-ring 83 may be disposed on the base plate 72 side, not on the support plate 81 side.

[0051] The sample injection section 82 is formed in a cylindrical shape. The sample injection section 82 may be formed in a funnel shape. The inside of the sample injection section 82 communicates with the through-hole 81a of the support plate 81. The sample injection section 82 may be integrated with the support plate 81.

[0052] The connecting portion 90 includes, for example, a male screw portion (not shown) provided on one of the base plate 72 and the support plate 81, and a female screw portion (not shown) provided on the other of the base plate 72 and the support plate 81. With this connecting portion 90, the filter lower portion 70 and the filter upper portion 80 can be connected by rotating the base plate 72 and the support plate 81 relatively. The connecting portion 90 may have a magnet and connect the base plate 72 and the support plate 81 by magnetic force.

[0053] Filtering of the sample 100 by the bacteria collection system 2 configured as above (step S1 shown in FIG. 4) is performed as follows. First, the sample 100 is placed in the sample injection section 82. Next, the inside of the filtrate collection container 71 is depressurized by the depressurization suction tube 73. Then, the sample 100 placed in the sample injection section 82 passes through the collection filter 101 due to the depressurization of the filtrate collection container 71, and the liquid content is collected in the filtrate collection container 71 as filtrate 102. At that time, turbid matter (solid content) in the sample 100 is trapped (collected) by the collection filter 101 in accordance with the filtration diameter of the collection filter 101.

[0054] Next, a method for recovering the collection filter 101 after filtering will be described. First, as shown in Fig. 6(a), the filter upper part 80 of the bacteria recovery system 2 is removed. At this time, it is advisable to also remove the connection part 90 as necessary.

[0055] Next, the lid 30 is removed from the container 10 shown in Fig. 2 described above, and the lid 30 is grasped, and as shown in Fig. 6(a), the filter fixing part 50 provided on the shaft body 40 is pressed against the collection filter 101 and hooked onto the collection filter 101 (step S2). Specifically, the tip 51 of the shaft body 40 is pierced into the collection filter 101, thereby fixing (hooking) one end of the collection filter 101 to the shaft body 40.

[0056] Next, as shown in Fig. 6(b) and Fig. 6(c), the shaft body 40 is rotated to wind the collection filter 101 around the shaft body 40 (step S3). Specifically, the collection filter 101 is rolled up while rotating the shaft body 40 in a state where it is close to the base plate 72. Thereafter, when the collection filter 101 is rolled up to the other end (the end opposite to the end pierced by the tip 51), the collection filter 101 is pierced by the above-mentioned tip 51 or a separately provided tip 61, thereby fixing the other end of the collection filter 101 to the shaft body 40. Note that the number of turns around the shaft body 40 varies depending on the size of the collection filter 101, so it is sufficient to wind it with a suitable number of turns at any time.

[0057] Here, since the peaks 51, 61 are provided with the convex portions 52, 62, it is possible to prevent the pierced collection filter 101 from falling or rewinding. Furthermore, since the base plate 72 is provided with the expanded diameter portion 72b, it becomes easier for the filter fixing portion 50 or the second filter fixing portion 60 including the peaks 51, 61 and the convex portions 52, 62 to penetrate the collection filter 101. Furthermore, while FIG. 6 shows an example in which the collection filter 101 is pierced into the peak 61, if there is no expanded diameter portion 72b when the collection filter 101 is pierced into the peak 61, it is not necessary to pierce the collection filter 101 into the peak 61.

[0058] Fig. 7 is a vertical cross-sectional view of the container 10 in which the collection filter 101 according to the first embodiment has been collected. Fig. 8 is a cross-sectional view taken along the line VIII-VIII shown in Fig. 7. 7 contains culture solution 103. Culture solution 103 is a liquid medium and serves as a nutrient source for culturing the bacteria or fungi captured by collection filter 101. Culture solution 103 may be included in culture kit 11 including collection filter 101 and container 10, or may be prepared separately from culture kit 11.

[0059] After the collection filter 101 is collected as described above, the culture solution 103 (liquid medium) is added to the container body 20 (step S4). Next, the shaft body 40 around which the collection filter 101 is wound is inserted into the container body 20 containing the culture solution 103, and the opening 20a is sealed with the lid 30 (step S5). Then, the container 10 shown in FIG. 7 containing the collection filter 101 is placed in a shaking incubator, and the collected bacteria or fungi are shake-cultured (step S6). Alternatively, the bacteria or fungi may be placed in an incubator kept at a constant temperature and subjected to static culture. With the above, the filtering of the sample 100 and the collection of the collection filter 101 to cell culture in the bacteria collection system 2 are completed.

[0060] As described above, according to the container 10 of the first embodiment, since the filter fixing part 50 is provided on the shaft body 40, the collection filter 101 to which the cells are attached can be hooked on the shaft body 40 and wound around the shaft body 40 for collection. In this way, the operator can collect the collection filter 101 with a simple operation. Therefore, when the collection filter 101 is placed in the container 10 for culture, the operator other than the container 10 for culture may accidentally touch the collection filter 101 to an unintended part, and the collection filter 101 may be contaminated by bacteria attached to the unintended part. In addition, the presence of the filter fixing part 50 makes it possible to prevent the collection filter 101 from falling off, regardless of the skill of the operator. In addition, it is possible to perform shaking culture with the shaft body 40 inserted in the container body 20, and the risk of the culture medium spilling from the container body 20 during shaking culture and the risk of contamination due to the introduction of unintended bacteria from the outside are reduced.

[0061] In this manner, the container 10 of the first embodiment described above comprises a container body 20 having an opening 20a, a lid 30 that is detachably attached to the container body 20 and seals the opening 20a, a shaft 40 that is provided on the lid 30 and extends toward the inside of the container body 20, and a filter fixing part 50 that is provided on the shaft 40 and hooks and fixes the collection filter 101 to the shaft 40. This configuration makes it possible to reduce the risk of contamination and operational errors regardless of the skill of the operator.

[0062] Furthermore, in this embodiment, the filter fixing part 50 has a tip 51 that extends in a direction other than the axial direction along which the shaft body 40 extends. With this configuration, the collection filter 101 can be easily fixed (hooked) by piercing the tip 51 of the shaft body 40 into the collection filter 101.

[0063] Furthermore, this embodiment includes a second filter fixing part 60 provided at a different position from the filter fixing part 50 in the circumferential direction of the shaft body 40. With this configuration, even if the filter fixing part 50 is not located at the winding start or winding end position of the collection filter 101, the second filter fixing part 60 can pierce the collection filter 101, thereby preventing the collection filter 101 from unwinding.

[0064] Furthermore, in this embodiment, the second filter fixing part 60 has a tip 61 that extends in a direction other than the axial direction along which the shaft body 40 extends. With this configuration, by piercing the tip 61 of the shaft body 40 into the collection filter 101, unwinding of the collection filter 101 can be more reliably suppressed.

[0065] Furthermore, in this embodiment, the apex 51, 61 has convex parts 52, 62 that extend in a direction other than the direction in which the apex 51, 61 extends. With this configuration, the collection filter 101 pierced by the apex 51, 61 is caught by the convex parts 52, 62, thereby reducing the risk of the collection filter 101 falling off. Also, as shown in Fig. 8, for example, the convex parts 62 act as spacers that form gaps in the overlapping parts caused by the rolled up collection filter 101, so that a flow of the culture solution 103 can be formed in the gaps in the rolled up collection filter 101, making it possible to effectively carry out cell culture. In consideration of this purpose, in the above example, one protrusion 52, 62 is formed on each of the peaks 51, 61, but it is not necessary that there is only one protrusion 52, 62 on each of the peaks 51, 61, and it is also possible to form multiple protrusions 52, 62 on each of the peaks 51, 61.

[0066] Moreover, the culture kit 11 of the present embodiment includes a collection filter 101 for collecting cells, and a container 10 in which the collection filter 101 and a culture solution 103 are sealed. According to this configuration, the collection filter 101 is housed in the container 10, and cell culture is possible.

[0067] Furthermore, the nucleic acid analysis system 1 of this embodiment collects cells using the container 10, encloses a culture medium in the container 10 and cultures the cells, or extracts nucleic acids from the cells collected and cultured using the culture kit 11, and analyzes the extracted nucleic acids. With this configuration, the risk of contamination is reduced, improving the accuracy of nucleic acid analysis.

[0068] Furthermore, in the collection filter recovery method of the present embodiment, the collection filter 101 to which cells are attached is hooked onto the shaft body 40 and wound around the shaft body 40 for recovery. With this configuration, it is not dependent on the skill of the operator, and it is possible to reduce the risk of contamination and operational errors.

[0069] Furthermore, in the cell culture method of this embodiment, the collection filter 101 with cells attached thereto is hooked onto the shaft body 40, wound around the shaft body 40, and collected, and the shaft body 40 with the collection filter 101 wound around it is immersed in the culture solution 103, and the cells are cultured by shaking. This is not dependent on the skill of the operator, and it is possible to reduce the risk of contamination and operational errors. Furthermore, it is possible to efficiently culture the cells attached to the collection filter 101 by shaking.

[0070] Second Embodiment Next, a second embodiment of the present invention will be described. In the following description, the same or equivalent configurations as those in the above-described embodiment are given the same reference numerals, and the description thereof will be simplified or omitted.

[0071] Fig. 9 is a vertical cross-sectional view of the container 10 according to the second embodiment. Fig. 10 is a vertical cross-sectional view of the shaft body 40 according to the second embodiment. As shown in FIG. 9, a filter fixing portion 50 of the second embodiment has a slit piece 53 in which a slit 53a is formed.

[0072] The slit piece 53 is, for example, an elastically deformable film member, and is attached to the peripheral surface 40a of the shaft body 40. As shown in Fig. 10, a mounting groove 40b is formed in the peripheral surface 40a of the shaft body 40, the mounting groove 40b having a depth corresponding to the radial thickness of the slit piece 53. A plurality of press-fit pins 43 are formed in the mounting groove 40b, rising radially from the bottom surface, at intervals in the axial direction. As shown in Fig. 9, the press-fit pins 43 are press-fitted into mounting holes 53b provided in the slit piece 53, and fix the slit piece 53 to the shaft body 40.

[0073] The slit piece 53 extends from the shaft 40 along the tangential direction of the circumferential surface 40a. A plurality of slits 53a are formed at intervals in the axial direction at the tip of the slit piece 53 extending in the tangential direction. The plurality of slits 53a extend in parallel from the tip of the slit piece 53 toward the fixed position of the slit piece 53. In the second embodiment, two slits 53a are formed in parallel, and the tip of the slit piece 53 is separated into three elastically deformable tongue portions. Hereinafter, a pair of tongue portions arranged on the axially outer sides of the two slits 53a is referred to as a first tongue portion 53A. Also, the tongue portion sandwiched between the two slits 53a is referred to as a second tongue portion 53B.

[0074] Fig. 11 is an explanatory diagram showing how the collection filter 101 is collected from the bacteria collection system 2 according to the second embodiment. Fig. 12 is a view taken along the arrow A shown in Fig. 11(a). As shown in FIGS. 11(a) and 12, in the second embodiment, the end of the collection filter 101 is hooked on the slit 53a of the slit piece 53, and the collection filter 101 is collected.

[0075] Specifically, as shown in Fig. 12, an end of the collection filter 101 is inserted into two slits 53a of the slit piece 53. Then, the end of the collection filter 101 is sandwiched between a first tongue portion 53A and a second tongue portion 53B separated by the two slits 53a. Note that in order to make it easier to insert the second tongue portion 53B under the collection filter 101, the length of the second tongue portion 53B may be longer than the first tongue portion 53A. Also, in order to make it easier to insert the second tongue portion 53B under the collection filter 101, the first tongue portion 53A may be bent upward (e.g., curled) in advance.

[0076] 11(b), the collection filter 101 fixed (hooked) to the shaft body 40 is lifted, and as in the first embodiment described above, the shaft body 40 is rotated and the collection filter 101 is wound around the shaft body 40. Thereafter, the collection filter 101 is rolled up to the other end (the end opposite the end inserted into the slit 53a), and then the collection filter 101 is pierced by the pointed portion 61 described above, thereby fixing the other end of the collection filter 101 to the shaft body 40.

[0077] Thus, the filter fixing part 50 of the second embodiment has a slit piece 53 in which a slit 53a is formed. With this configuration, by clamping and fixing an end of the collection filter 101 in the slit 53a, it becomes possible to collect the collection filter 101 while rolling it up, as in the first embodiment described above. Therefore, it is possible to obtain the same effects as those described in the first embodiment described above.

[0078] Third Embodiment Next, a third embodiment of the present invention will be described. In the following description, the same or equivalent configurations as those in the above-described embodiment are given the same reference numerals, and the description thereof will be simplified or omitted.

[0079] Fig. 13 is a vertical cross-sectional view of the container 10 according to the third embodiment. Fig. 14 is a cross-sectional view taken along line XIV-XIV shown in Fig. 13. As shown in Fig. 13, in the third embodiment, a plurality of filter fixing portions 50 and second filter fixing portions 60 are formed at intervals in the axial direction on the peripheral surface 40a of the shaft body 40. Although the apexes 51, 61 of the third embodiment shown in Figs. 13 and 14 do not have the above-mentioned protrusions 52, 62, they may have the above-mentioned protrusions 52, 62.

[0080] Moreover, the shaft body 40 of the third embodiment is a hollow shaft. A hollow space 44 is formed in the shaft body 40, penetrating from the upper end 41 to the lower end 42. A plurality of through holes 45 are formed in the peripheral surface 40a of the shaft body 40. The through holes 45 communicate with the hollow space 44 of the shaft body 40. The plurality of through holes 45 are formed at intervals in the axial direction of the shaft body 40. Moreover, the through holes 45 are formed at two different positions in the circumferential direction of the shaft body 40, as shown in FIG.

[0081] According to the third embodiment having the above configuration, the culture solution 103 that has entered the hollow space 44 from the lower end 42 of the shaft body 40 can be made to flow out from the through-hole 45 of the shaft body 40. This allows the culture solution 103 to be supplied to the collection filter 101 wrapped around the shaft body 40 from the radially inner side as well, enabling efficient shaking culture of the cells attached to the collection filter 101.

[0082] Although the preferred embodiment of the present invention has been described above with reference to the drawings, the present invention is not limited to the above embodiment. The shapes and combinations of the components shown in the above embodiment are merely examples, and various modifications can be made based on design requirements, etc., without departing from the spirit of the present invention.

[0083] For example, modifications as shown in FIGS. 15 and 16 may be adopted.

[0084] FIG. 15 is a cross-sectional view of a container 10 according to a modified example of the first embodiment. As shown in FIG. 15, the points 51, 61 may extend in an oblique direction (for example, a tangential direction of the shaft 40) other than the radial direction perpendicular to the central axis O of the shaft 40.

[0085] FIG. 16 is a cross-sectional view of a container 10 according to a modified example of the first embodiment. As shown in FIG. 16, the tips 51, 61 do not necessarily extend linearly in the radial direction perpendicular to the central axis O of the shaft body 40, but may extend curvedly in the circumferential direction of the shaft body 40.

[0086] 1. Nucleic Acid Analysis System 10 containers 11 Culture Kit 20 Container body 20a opening 30 Lid 40 Axial body 50 Filter fixing part 51 Apex 52 Convex 61 Apex 62 Convex 100 samples 101 Collection filter (filter) 103 Culture solution

Claims

1. A container body having an opening; a cover body that is detachably attached to the container body and seals the opening; A shaft body provided on the lid body and extending toward the inside of the container body; a filter fixing portion provided on the peripheral surface of the shaft body and having a hook portion for hooking a filter that is wound around and fixed to the shaft body.

2. The container according to claim 1 , wherein the hook portion is a pointed portion extending in a direction other than the axial direction along which the shaft body extends.

3. The container according to claim 1 , wherein the hook portion is a slit piece having a slit formed therein.

4. The container according to any one of claims 1 to 3, further comprising a second filter fixing part provided at a different position from the filter fixing part in the circumferential direction of the shaft.

5. The container according to claim 4 , wherein the second filter fixing portion has a tip extending in a direction other than the axial direction along which the shaft body extends.

6. The container of claim 2 or 5, wherein the pointed portion has a protrusion extending in a direction other than the direction in which the pointed portion extends.

7. A filter for collecting cells; A container according to any one of claims 1 to 6 in which the filter and the culture solution are enclosed; A culture kit comprising:

8. A nucleic acid analysis system comprising: collecting cells using the container according to any one of claims 1 to 6; enclosing a culture medium in the container; extracting nucleic acid from the cultured cells; and analyzing the extracted nucleic acid.

9. A nucleic acid analysis system, which extracts nucleic acid from cells collected and cultured using the culture kit according to claim 7, and analyzes the extracted nucleic acid.

10. A method of attaching the filter to the hook portion of the shaft provided on the lid removed from the container according to any one of claims 1 to 6, and rotating the shaft to wrap the filter around the shaft and secure it thereto; the shaft body around which the filter is wound and fixed is inserted into the container body to which a culture medium has been added, and the opening of the container body is sealed with the lid body to culture the cells captured on the filter. Cell culture method.

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