JP7635554B2 - Container, culture kit, nucleic acid analysis system, and cell culture method - Google Patents

Description

The present invention relates to a container, a culture kit, a nucleic acid analysis system , and a cell culture method .

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 that includes a funnel and a filtration bottle, and collects the sample by filtering the liquid containing the sample poured into the collection section.

International Publication No. 2019/043779

After collecting the sample on the membrane filter, it is retrieved using tweezers and transferred to a Petri dish for cell culture. However, when picking up the membrane filter with the tweezers, depending on the operator's skill, there is a risk that the membrane filter may be accidentally dropped onto the floor or other surfaces. In addition, if the tweezers are used multiple times, the sample attached to the tweezers may adhere to other membrane filters, resulting in contamination.

The present invention has been made in consideration of the above-mentioned circumstances, and aims to provide a container, a culture kit, a nucleic acid analysis system , and a cell culture method that are not dependent on the skill of the operator and that can reduce the risk of contamination and operational errors.

(1) A container according to one aspect of the present invention includes 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 part that is attached to the shaft body and hooks a filter and fixes it to the shaft body.

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

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

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

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

(6) In the container described in (1) above, the tip may have a protrusion extending in a direction other than the direction in which the tip extends.

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

(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.

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

According to one aspect of the present invention, a container, a culture kit, and a nucleic acid analysis system can be obtained that are not dependent on the skill of the operator and that can reduce the risk of contamination and operational errors.

1 is a schematic diagram of a nucleic acid analysis system according to a first embodiment. FIG. 2 is a vertical cross-sectional view of a container used in the bacteria collection system according to the first embodiment. 3 is a cross-sectional view taken along the line III-III of FIG. 2. FIG. 2 is a process diagram from sample collection to culture according to the first embodiment. FIG. 1 is a vertical cross-sectional view of a bacteria collection system according to a first embodiment. FIG. 4 is an explanatory diagram showing a state in which a collection filter is collected from the bacteria collection system according to the first embodiment. FIG. 2 is a vertical cross-sectional view of a container in which the collection filter according to the first embodiment is collected. 8 is a cross-sectional view taken along the line VIII-VIII of FIG. 7. FIG. 11 is a vertical cross-sectional view of a container according to a second embodiment. FIG. 11 is a longitudinal sectional view of a shaft body according to a second embodiment. FIG. 11 is an explanatory diagram showing a state in which a collection filter is collected from a bacteria collection system according to a second embodiment. FIG. 11B is a view taken along the arrow A in FIG. FIG. 11 is a vertical cross-sectional view of a container according to a third embodiment. 14 is a cross-sectional view taken along the line XIV-XIV of FIG. 13. FIG. 4 is a cross-sectional view of a container according to a modified example of the first embodiment. FIG. 4 is a cross-sectional view of a container according to a modified example of the first embodiment.

Below, the container, culture kit, and nucleic acid analysis system according to the embodiments of the present invention will be described in detail with reference to the drawings. Below, 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.

〔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, which uses a filter funnel and a suction system to pass a liquid through a membrane filter set in the filter funnel and collect a sample such as microorganisms contained in the liquid on the surface of the membrane filter.

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

However, when using tweezers to retrieve membrane filters, the following problems arise. For example, there is a risk of accidentally dropping the membrane filter when picking it up with the tweezers. In addition, if 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 a membrane filter with tweezers is itself cumbersome.

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. Also, 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.
In addition, as described below, when using a liquid analysis system to analyze whether a liquid sample contains bacteria (bacteria or fungi), 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.

In an 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. The shaft body with the collection filter wound around it is then 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. Furthermore, efficient culture is possible in a liquid culture medium using a collection filter. Furthermore, efficient culture is possible by shaking the liquid culture medium containing the collection filter.

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.

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 may be the produced beverage, the water used to produce the beverage, or a liquid in the process of producing the beverage. Alternatively, the sample 100 may be a liquid in which microorganisms have been collected from a swab or the like used to wipe the testing environment in order to test for the presence or absence of bacterial contamination in the production environment and the level of contamination.

The bacteria can be collected, for example, by applying pressure or vacuum to the collected liquid and filtering through 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, the liquid in which the bacteria are collected by centrifugation or the like, or the 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.

The nucleic acid extraction system 3 is a system that destroys (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 also be nucleic acids to which a site that exhibits fluorescence, luminescence, or quenching effects under specific conditions has been added in order to detect them in the detection process (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.

The hybridization reaction system 4 is a system that causes a hybridization reaction of the nucleic acid in the sample 100. In this process, the sample 100 is heated, for example, to 60°C and stirred to cause a hybridization reaction that matches the other nucleic acid described above. In this reaction, for example, a site that has a fluorescent, luminescent, or quenching effect under specific conditions that 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.

The structure of the other nucleic acid described above can be designed to react with a specific nucleic acid, so that it can react only with the nucleic acid contained in, for example, a specific bacterium or fungus 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 or fungus.

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

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

By using a series of systems as described above, the nucleic acid analysis system 1 analyzes whether a particular microorganism (such as bacteria or fungi) is present in the sample 100 or its concentration.

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 the 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.

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, the direction that intersects with the central axis O is referred to as the radial direction. The direction that rotates around the central axis O is referred to as the circumferential direction.

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 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.

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.

The top wall 31 is formed in a disk shape and is connected to the upper end of the peripheral wall 32, closing 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 at 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 that are combined by fitting them together, and may be one piece.

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. Meanwhile, the lower end 42 of the shaft body 40 does not contact the bottom 22 of the container body 20, but extends close to 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.

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 the collection filter 101 (see Figures 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 Figure 3.

The tip 51 extends in a radial direction that intersects the central axis O at a right angle. The tip 51 is formed in a cone shape that tapers toward the outside in the radial direction. The tip 51 may be formed in a needle shape that is sharp only at the tip. The tip 51 may also extend in a direction other than the axial direction in which the shaft body 40 extends, for example, in an oblique direction that intersects the central axis O at an acute or obtuse angle.

The tip 51 further has a protrusion 52 extending in a direction other than the direction in which the tip 51 extends. The protrusion 52 is formed in a plate shape extending from approximately the middle of the slope of the conical tip 51 in a direction perpendicular to the direction in which the tip 51 extends. The protrusion 52 may be an elbow shape bent in an L-shape with respect to the tip 51, or a hook shape like a fish hook, as long as it is in a direction other than the direction in which the tip 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 tip 51 extends.

In the first embodiment, the shaft body 40, the tip 51, and the protrusion 52 are integrally molded, but they may be assembled by fitting or other means of separate parts. For example, the protrusion 52 may be a ring member that fits skewer-like into the tip 51. The tip 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 tetrahedron.

As shown in FIG. 3, a second filter fixing part 60 is provided on the peripheral surface 40a of the shaft body 40 at a different position in the circumferential direction from the filter fixing part 50. The second filter fixing part 60 has a peak 61 and a protrusion 62, and has a similar configuration to the filter fixing part 50 described above. 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.

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 the 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 the state in which the collection filter 101 is collected from the bacteria collection system 2 according to the first embodiment.

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 portion 70 and an upper filter portion 80 that sandwich the collection filter 101 from above and below. The collection filter 101 is formed in a circular shape in a plan view. The lower filter portion 70 and the upper filter portion 80 are connected via a connecting portion 90 in the axial direction along a central axis O1 that passes through the center of the collection filter 101 as a common axis.

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 body such as a vacuum pump (not shown) is connected to the reduced pressure suction tube 73.

The base plate 72 is formed with a plurality of through holes 72a that communicate with the inside of the filtrate collection container 71. Some of the through holes 72a are formed with enlarged diameter sections 72b. The enlarged diameter sections 72b are grooves with a larger diameter than the through holes 72a. The enlarged diameter sections 72b have a diameter and depth that allow the tips 51, 61 and protrusions 52, 62 of the filter fixing part 50 and the second filter fixing part 60 described above to be inserted. The enlarged diameter sections 72b may be formed on the upper surface of the base plate 72 as independent grooves that do not communicate with the through holes 72a.

A filter positioning portion 72c is formed on the upper surface of the base plate 72, surrounding the multiple through holes 72a and the enlarged diameter portion 72b. 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.

The filter positioning portion 72c is an annular protrusion provided on the upper surface of the base plate 72 at 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 also be a groove that is recessed relatively to the upper surface of the base plate 72.

The filter upper portion 80 includes a support plate 81 having a through hole 81a formed therein that penetrates in the axial direction along which the central axis O1 extends, and a cylindrical sample injection portion 82 that is attached to the support plate 81 and whose inside communicates 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 in between.

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 provided 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 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.

The sample injection section 82 is formed in a cylindrical shape. The sample injection section 82 may also be formed in a funnel shape. The inside of the sample injection section 82 is connected to the through-hole 81a of the support plate 81. The sample injection section 82 may also be integrated with the support plate 81.

The connecting portion 90 includes, for example, a male thread portion (not shown) provided on one of the base plate 72 and the support plate 81, and a female thread 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 relative to each other. The connecting portion 90 may have a magnet and connect the base plate 72 and the support plate 81 by magnetic force.

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 vacuum 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, the turbid matter (solid content) in the sample 100 is trapped (collected) by the collection filter 101 according to the filtration diameter of the collection filter 101.

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 connecting part 90, if necessary.

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

6(b) and 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. After that, when the collection filter 101 is rolled up to the other end (the end opposite the end pierced by the tip 51), the collection filter 101 is pierced by the tip 51 described above 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 an appropriate number of turns at the appropriate time.

Here, the peaks 51, 61 are provided with the convex portions 52, 62, so that the pierced collection filter 101 can be prevented from falling or rewinding. In addition, the base plate 72 is provided with the enlarged diameter portion 72b, so that the filter fixing portion 50 or the second filter fixing portion 60 including the peaks 51, 61 and the convex portions 52, 62 can easily penetrate the collection filter 101. In addition, while FIG. 6 shows an example in which the collection filter 101 is pierced into the peak 61, if there is no enlarged 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.

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 line VIII-VIII shown in Fig. 7.
7 contains a culture solution 103. The culture solution 103 is a liquid medium and serves as a nutrient source for culturing the bacteria or fungi captured by the collection filter 101. The culture solution 103 may be included in a culture kit 11 including the collection filter 101 and the container 10, or may be prepared separately from the culture kit 11.

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 with the collection filter 101 wound around it is inserted into the container body 20 containing the culture solution 103, and the opening 20a is sealed with the lid body 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 and fungi are shake-cultured (step S6). Alternatively, they may be placed in an incubator kept at a constant temperature and subjected to static culture. This completes the process from filtering the sample 100 and collecting the collection filter 101 to cell culture in the bacteria collection system 2.

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 placing the collection filter 101 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 with 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, which reduces 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.

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

In addition, in this embodiment, the filter fixing portion 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.

In addition, in this embodiment, a second filter fixing part 60 is 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 start and end positions of the collection filter 101, the second filter fixing part 60 can pierce the collection filter 101 and prevent the collection filter 101 from rewinding.

In addition, in this embodiment, the second filter fixing portion 60 has a tip 61 that extends in a direction other than the axial direction in which the shaft body 40 extends. With this configuration, by piercing the tip 61 of the shaft body 40 into the collection filter 101, it is possible to more reliably suppress the rewinding of the collection filter 101.

Furthermore, in this embodiment, the apex 51, 61 has a protrusion 52, 62 that extends 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 protrusion 52, 62, reducing the risk of the collection filter 101 falling. Also, as shown in Fig. 8, for example, the protrusion 62 acts as a spacer that forms a gap in the overlapping portion caused by the rolled up collection filter 101, so that a flow of the culture solution 103 can be formed in the gap 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 convex portion 52, 62 is formed on each of the peaks 51, 61, but it is not necessary that there is only one convex portion 52, 62 on each of the peaks 51, 61, and it is also possible to form multiple convex portions 52, 62 on each of the peaks 51, 61.

The culture kit 11 of this embodiment also includes a collection filter 101 for collecting cells, and a container 10 in which the collection filter 101 and culture solution 103 are sealed. With this configuration, the collection filter 101 is housed in the container 10, making it possible to culture cells.

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 cells collected and cultured using the culture kit 11, and analyzes the extracted nucleic acids. This configuration reduces the risk of contamination, improving the accuracy of nucleic acid analysis.

In addition, the collection filter recovery method of this embodiment involves hooking the collection filter 101 with cells attached to the shaft body 40 and winding it around the shaft body 40 to recover it. This configuration makes it possible to reduce the risk of contamination and operational errors without being dependent on the skill of the operator.

In addition, 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. In addition, it is possible to efficiently culture the cells attached to the collection filter 101 by shaking.

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

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.

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 on the peripheral surface 40a of the shaft body 40, with a depth corresponding to the radial thickness of the slit piece 53. The mounting groove 40b has a plurality of press-fit pins 43 that rise radially from the bottom surface and are formed 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.

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 the tip of the slit piece 53 extending in the tangential direction, spaced apart in the axial 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, the pair of tongue portions arranged axially outside the two slits 53a will be referred to as the first tongue portion 53A. The tongue portion sandwiched between the two slits 53a will be referred to as the second tongue portion 53B.

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 in Fig. 11(a).
As shown in FIGS. 11A and 12, in the second embodiment, the collection filter 101 is collected by hooking an end of the collection filter 101 onto a slit 53 a of a slit piece 53 .

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

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

In this way, 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 the 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.

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

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. Note that the pointed portions 51, 61 of the third embodiment shown in Figs. 13 and 14 do not have the above-mentioned convex portions 52, 62, but may have the above-mentioned convex portions 52, 62.

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 are connected to the hollow space 44 of the shaft body 40. The through holes 45 are formed at intervals in the axial direction of the shaft body 40. The through holes 45 are formed at two different locations in the circumferential direction of the shaft body 40, as shown in FIG. 14.

According to the third embodiment having the above configuration, the culture fluid 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 fluid 103 to be supplied from the radial inside to the collection filter 101 wrapped around the shaft body 40, enabling efficient shaking culture of the cells attached to the collection filter 101.

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.

For example, modified examples such as those shown in Figures 15 and 16 may be adopted.

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

FIG. 16 is a cross-sectional view of the 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 .

1 Nucleic acid analysis system 10 Container 11 Culture kit 20 Container body 20a Opening 30 Lid 40 Shaft 50 Filter fixing part 51 Tip 52 Convex part 61 Tip 62 Convex part 100 Sample 101 Collection filter (filter)
103 Culture medium

Claims (10)

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 . 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. The container according to claim 1 , wherein the hook portion is a slit piece having a slit formed therein. The container according to any one of claims 1 to 3, comprising a second filter fixing part provided at a different position from the filter fixing part in the circumferential direction of the shaft. The container according to claim 4, wherein the second filter fixing portion has a tip that extends in a direction other than the axial direction along which the shaft body extends. The container according to 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. 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: A nucleic acid analysis system that collects cells using a container according to any one of claims 1 to 6, encloses a culture medium in the container, extracts nucleic acid from the cultured cells, and analyzes the extracted nucleic acid. A nucleic acid analysis system that extracts nucleic acid from cells collected and cultured using the culture kit according to claim 7, and analyzes the extracted nucleic acid. The filter is hooked to the hook portion of the shaft provided on the lid body removed from the container according to any one of claims 1 to 6, and the shaft body is rotated to wind and fix the filter around the shaft body;
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 methods.

Images