JP2018061451A - Nucleic acid extraction device and nucleic acid extraction method - Google Patents

Abstract

There is provided a nucleic acid extraction apparatus and the like capable of inspecting an object to be measured with high sensitivity and high accuracy without using a large amount of nucleic acid inspection reagent. A nucleic acid extraction apparatus (100) includes a container (10), a first injection port (21) for injecting a sample stock solution into the container (10), and an object to be measured contained in the sample stock solution injected from the first injection port (21). A capture unit 30 for capturing by filtration; a second inlet 22 for injecting a nucleic acid extraction reagent for extracting nucleic acid from the measurement object captured by the capture unit 30 into the container 10; The retention part 40 in which the nucleic acid solution containing the nucleic acid extracted from the measurement object is retained, the heating part 50 for heating the nucleic acid solution retained in the retention part 40, and the nucleic acid solution is heated by the heating part 50 A first discharge port 61 for discharging the vapor of the solvent generated during the operation from the container 10, and the first discharge port 61 is for exhausting the air in the container 10 when the inside of the container 10 is decompressed Also serves as an exhaust port. [Selection] Figure 2

Description

  The present invention relates to a nucleic acid extraction apparatus and a nucleic acid extraction method for extracting nucleic acid (deoxyribonucleic acid, ribonucleic acid) from an object to be measured contained in a sample stock solution.

  Nucleic acid amplification methods such as polymerase chain reaction (PCR) are used in the inspection of microorganisms (measurement objects) such as bacteria contained in beverages and the like. The nucleic acid amplification method has an advantage that the inspection process can be greatly speeded up and simplified as compared with the culture method. As a result, microorganisms present in the sample stock solution such as a beverage can be rapidly detected.

  In order to perform a test using the nucleic acid amplification method, a pretreatment for extracting nucleic acid from a microorganism contained in a sample stock solution is required before nucleic acid amplification. Conventionally, as such pretreatment, a sample stock solution is filtered with a filter to capture microorganisms contained in the sample stock solution, and nucleic acids are extracted from the microorganisms with a nucleic acid extraction reagent to obtain a nucleic acid solution containing nucleic acids. A method is known (for example, Patent Document 1).

JP-A-4-36197

  In order to perform a test with high sensitivity by the PCR method or the like, it is necessary to obtain a nucleic acid solution at a high nucleic acid concentration from a sample stock solution containing microorganisms.

  In the conventional nucleic acid extraction method (pretreatment), microorganisms in the sample stock solution are captured by a filter. Usually, a filter having a certain filtration area is used in order to shorten the working time. Thereafter, by using a large amount of nucleic acid extraction reagent corresponding to the filtration area of the filter, nucleic acid is extracted from the captured microorganism to obtain a nucleic acid solution. Then, a part of the nucleic acid solution (for example, 1% to 10%) is weighed and collected, and a nucleic acid test reagent is added thereto to obtain a test solution for use in a test such as a PCR method. For example, when 100 μl of the test solution is obtained, 10 μl of 100 μl of the nucleic acid solution is collected, and a nucleic acid test reagent is added thereto to obtain 100 μl of the test solution. The remaining 90 μl of the nucleic acid solution is discarded.

  However, in such a method, since only a part of the nucleic acid solution is used, as a result, the concentration of the test solution is lowered, and the detection sensitivity during the test is lowered. Moreover, since a nucleic acid extraction reagent is expensive, using it in large quantities contributes to an increase in cost.

  Therefore, if an attempt is made to obtain a test solution using a large amount of nucleic acid test reagent in order to increase detection sensitivity, a large amount of expensive nucleic acid test reagent is required, which further increases the cost. Moreover, the test solution more than necessary may be prepared.

  In addition, when a small amount of nucleic acid extraction reagent is injected, the nucleic acid extraction reagent does not spread evenly on the filter, and the nucleic acid cannot be accurately extracted from the microorganisms captured by the filter. On the other hand, if the area of the filter is reduced in response to a small amount of nucleic acid extraction reagent, the filtration time becomes longer, and the time for extracting nucleic acid from the microorganisms contained in the sample stock solution becomes longer. For this reason, it is impossible to meet the work time required by the user.

  Naturally, if the sample stock solution is reduced to reduce the filtration time, the amount of microorganisms (measurement object) captured from the sample stock solution is reduced. As a result, the concentration of the test solution is lowered and the detection sensitivity is lowered.

  The present invention has been made in order to solve such problems, and a nucleic acid extraction apparatus and a nucleic acid that can inspect an object to be measured with high sensitivity and high accuracy without using a large amount of a nucleic acid inspection reagent. An object is to provide an extraction method.

  In order to achieve the above object, according to one aspect of the nucleic acid extraction apparatus of the present invention, a container, a first injection port for injecting a sample stock solution into the container, and at least a part thereof are arranged in the container. A capture unit for capturing the measurement object contained in the sample stock solution injected from the first injection port by filtration; and a nucleic acid extraction for extracting nucleic acid from the measurement object captured by the capture unit A second injection port for injecting a reagent into the container; a retention part provided in the container for retaining a nucleic acid solution containing a nucleic acid extracted from the object to be measured; and the retention part remaining in the retention part A heating unit for heating the nucleic acid solution; and a first discharge port for discharging the vapor of the solvent generated when the nucleic acid solution is heated by the heating unit from the container, wherein the first discharge port is , When depressurizing the inside of the container Characterized in that it also serves as an exhaust port for exhausting the air in the vessel.

  Also, one aspect of the nucleic acid extraction method according to the present invention is to suck the sample stock solution injected into the container by evacuating the air in the container from a discharge port provided in the container and depressurizing the inside of the container. The sample to be measured is captured from the sample stock solution at the capture unit by filtration, the nucleic acid extraction reagent is injected into the container, the nucleic acid is extracted from the sample to be captured at the capture unit, and the extracted nucleic acid is The contained nucleic acid solution is retained in the retention part through the capturing part, the nucleic acid solution retained in the retention part is heated under reduced pressure, and the solvent vapor of the nucleic acid solution is discharged from the outlet to the outside of the container. Then, the nucleic acid solution is concentrated.

  According to the present invention, it is possible to inspect an object to be measured with high sensitivity and high accuracy without using a large amount of nucleic acid inspection reagent.

1 is a perspective view schematically showing a nucleic acid extraction apparatus according to Embodiment 1. FIG. 2 is a cross-sectional view of a nucleic acid extraction unit according to Embodiment 1. FIG. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when inject | pouring a sample stock solution into a container. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows the to-be-measured object captured by the capture part. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when inject | pouring a nucleic acid extraction reagent into a container. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when a nucleic acid extraction reagent is hold | maintained at a capture part. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when moving a nucleic acid solution from a 1st container part to a 2nd container part. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when a nucleic acid solution is made to retain in a retention part. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when heating the nucleic acid solution made to stay in a retention part. In the nucleic acid extraction method which concerns on Embodiment 1, it is a figure which shows a mode when discharging | emitting a nucleic acid solution from a container. FIG. 6 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 1 of Embodiment 1. FIG. 5 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 2 of Embodiment 1. FIG. 10 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 3 of Embodiment 1. It is sectional drawing of the nucleic acid extraction unit used for the nucleic acid extraction apparatus which concerns on another example of the modification 3 of Embodiment 1. FIG. FIG. 10 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 4 of Embodiment 1. It is sectional drawing of the nucleic acid extraction unit used for the nucleic acid extraction apparatus which concerns on another example of the modification 4 of Embodiment 1. FIG. It is a figure which shows the structure of the support part of a capture part in FIG. 9, and a metal member. 6 is a perspective view schematically showing a nucleic acid extraction apparatus according to Embodiment 2. FIG. 6 is a cross-sectional view of a nucleic acid extraction unit according to Embodiment 2. FIG. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when inject | pouring a sample stock solution into a container. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows the to-be-measured object captured by the capture part. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when inject | pouring a nucleic acid extraction reagent into a container. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when a nucleic acid extraction reagent is hold | maintained at a capture part. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when moving a nucleic acid solution from a 1st container part to a 2nd container part. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when a nucleic acid solution is made to retain in a retention part. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when heating the nucleic acid solution made to retain in a retention part. In the nucleic acid extraction method which concerns on Embodiment 2, it is a figure which shows a mode when discharging | emitting a nucleic acid solution from a container. 6 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to a modification of the second embodiment. FIG. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is plugged) used for the nucleic acid extraction apparatus which concerns on Embodiment 3. FIG. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is opened) used for the nucleic acid extraction apparatus which concerns on Embodiment 3. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is plugged) used for the nucleic acid extraction apparatus which concerns on the modification 1 of Embodiment 3. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is opened) used for the nucleic acid extraction apparatus which concerns on the modification 2 of Embodiment 3. FIG. 10 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 2 of Embodiment 3. FIG. 11 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to a first example of modification 2 of the third embodiment. FIG. 10 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction apparatus according to a second example of modification 2 of the third embodiment. FIG. 10 is a cross-sectional view of a nucleic acid extraction unit used in a nucleic acid extraction device according to Modification 3 of Embodiment 3. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is plugged) used for the nucleic acid extraction apparatus which concerns on the modification 4 of Embodiment 3. It is sectional drawing of the nucleic acid extraction unit (when the 2nd discharge port is opened) used for the nucleic acid extraction apparatus which concerns on the modification 4 of Embodiment 3.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. Note that each of the embodiments described below shows a preferred specific example of the present invention. Therefore, numerical values, shapes, materials, components, arrangement positions and connection forms of components, steps (steps) and order of steps, and the like shown in the following embodiments are examples and limit the present invention. It is not the purpose to do. Therefore, among the constituent elements in the following embodiments, constituent elements that are not described in the independent claims showing the highest concept of the present invention are described as optional constituent elements.

  Each figure is a schematic diagram and is not necessarily illustrated strictly. Moreover, in each figure, the same code | symbol is attached | subjected to the substantially same structure, The overlapping description is abbreviate | omitted or simplified.

(Embodiment 1)
[Nucleic acid extraction equipment]
The configuration of the nucleic acid extraction apparatus 100 and the nucleic acid extraction unit 1 according to Embodiment 1 will be described with reference to FIGS. FIG. 1 is a perspective view schematically showing a nucleic acid extraction apparatus 100 according to the first embodiment. FIG. 2 is a cross-sectional view of the nucleic acid extraction unit 1 according to the first embodiment.

  As shown in FIG. 1, the nucleic acid extraction apparatus 100 is an apparatus for extracting nucleic acid from a measurement object contained in a sample stock solution, and includes a nucleic acid extraction unit 1, a vacuum pump 101, and a reaction box 102.

  An object to be measured contained in the sample stock solution (sample) is an object for extracting nucleic acid (object for nucleic acid extraction), and is, for example, a microorganism such as a bacterium, a virus, or a tissue cell. A sample stock solution containing microorganisms can be collected from a beverage, for example.

  Microorganisms such as bacteria, viruses or tissue cells are inspection objects to be inspected by nucleic acid amplification by PCR or the like. That is, the nucleic acid extraction apparatus 100 is an apparatus used for pretreatment before performing nucleic acid amplification, and the nucleic acid extracted by the nucleic acid extraction apparatus 100 is used for a desired test. In the present embodiment, the nucleic acid extracted by the nucleic acid extraction apparatus 100 is recovered as a liquid (nucleic acid solution) containing the extracted nucleic acid.

  The nucleic acid extraction unit 1 is a processing unit for performing a nucleic acid extraction process from a sample stock solution, and is installed in a reaction box 102 of a nucleic acid extraction apparatus 100 as shown in FIG. The nucleic acid extraction unit 1 is a replaceable cartridge and can be removed from the reaction box 102. That is, the nucleic acid extraction unit 1 can be attached to and detached from the reaction box 102. For example, the nucleic acid extraction unit 1 is replaced for each nucleic acid extraction process. FIG. 1 shows a state where the entire lid of the reaction box 102 is opened.

  As shown in FIG. 2, the nucleic acid extraction unit 1 includes a container 10, a first injection port 21, a second injection port 22, a capture unit 30, a retention unit 40, a heating unit 50, and a first discharge port. 61.

  The container 10 is a processing container for performing a nucleic acid extraction process from an object to be measured (microorganisms or the like) contained in the sample stock solution with the sample stock solution and the nucleic acid extraction reagent to be injected. The nucleic acid extracted from the sample stock solution is discharged from the container 10 as a nucleic acid solution (nucleic acid extract).

  In the present embodiment, the container 10 includes a first container part 11, a second container part 12, and a third container part 13. The first container part 11 is located in the upper part of the container 10, the second container part 12 is located in the middle part of the container 10, and the third container part 13 is located in the lower part (bottom part) of the container 10. . The first container part 11 and the second container part 12 divide the internal space of the container 10 with the capturing part 30 as a boundary. The container 10 may be divided into three or more, or may be integrated as an integrally molded product without being divided.

  The material of the container 10 is not particularly limited, but the container 10 may be made of a material having high thermal conductivity and heat resistance so that heat treatment can be performed. For example, the container 10 may be made of a metal material such as aluminum or stainless steel, a resin material having high heat resistance such as polypropylene (PP) or polycarbonate (PC), or an inorganic material such as glass or ceramic. When using resin as the material of the container 10, it is good to use high thermal conductivity resin with high thermal conductivity. The container 10 may be configured by combining the above materials. For example, the 1st container part 11, the 2nd container part 12, and the 3rd container part 13 may be comprised with a different material. In this case, in the present embodiment, since the third container part 13 functions as the heating part 50 as will be described later, at least the third container part 13 has a thermal conductivity such as a metal material such as aluminum or a high thermal conductive resin. It is good to be composed of a high material.

  The first inlet 21, the second inlet 22 and the second outlet 62 are provided in the container 10. Specifically, the first inlet 21, the second inlet 22, and the second outlet 62 are provided in the partition wall of the container 10.

  The first injection port 21 is a sample stock solution injection port for injecting a sample stock solution into the container 10. The first inlet 21 is provided on the upper wall of the first container portion 11. The first inlet 21 is a through hole that communicates the outside of the container 10 and the internal space of the container 10 (first container portion 11). The first injection port 21 is inserted with the injection tube of the sample insertion cup 200. As shown in FIG. 1, the sample stock solution is injected into the container 10 (first container portion 11) through the first injection port 21 from the sample insertion cup 200 installed in the nucleic acid extraction apparatus 100.

  The second injection port 22 is a nucleic acid extraction reagent injection port for injecting the nucleic acid extraction reagent into the container 10. The second inlet 22 is provided on the upper wall of the first container portion 11 in the same manner as the first inlet 21. The second inlet 22 is a through hole that communicates the outside of the container 10 and the internal space of the container 10 (first container portion 11).

  The nucleic acid extraction reagent injected from the second injection port 22 is a liquid reagent for extracting the nucleic acid from the measurement object captured by the capturing unit 30. For example, the nucleic acid extraction reagent functions to extract the nucleic acid from the cell membrane of the microorganism that is the measurement object. Have As shown in FIG. 1, the nucleic acid extraction reagent is injected from the nucleic acid extraction reagent container 300 installed in the nucleic acid extraction apparatus 100 into the first container portion 11 through the second injection port 22. Specifically, the nucleic acid extraction reagent is injected into the container 10 through an injection needle provided in the nucleic acid extraction reagent container 300.

  In the present embodiment, the second inlet 22 is provided with a rubber plug 22a that closes the second inlet 22 (through hole). The rubber stopper 22a is made of a rubber material that can be penetrated by the injection needle of the nucleic acid extraction reagent container 300. When the nucleic acid extraction reagent in the nucleic acid extraction reagent container 300 is injected into the first container portion 11, the injection needle of the nucleic acid extraction reagent container 300 is made to penetrate the rubber stopper 22a. Thereby, the nucleic acid extraction reagent can be injected into the container 10 from the nucleic acid extraction reagent container 300 through the second injection port 22. The rubber material of the rubber plug 22a is not particularly limited, but, for example, silicone rubber or fluorine rubber is used.

  The capturing unit 30 is a capturing unit that is at least partially disposed in the container 10 and captures the object to be measured included in the sample stock solution injected from the first injection port 21 by filtration. The capturing unit 30 includes a filter unit 31 (main body unit) for filtering and capturing an object to be measured, and a support unit 32 that supports the filter unit 31. The capturing unit 30 is fixed to the container 10 so that the filter unit 31 is located inside the container 10.

  The filter unit 31 is placed on a plate-like support unit 32. A plurality of through holes (eyes) are formed in the central portion of the support portion 32 corresponding to the filter portion 31. That is, the support part 32 is a eye plate. Further, the peripheral part of the support part 32 is sandwiched between the first container part 11 and the second container part 12. Thereby, the support part 32 is fixed to the container 10. The support part 32, the first container part 11, and the second container part 12 are fixed by, for example, four screws.

  The filter unit 31 is a filter having a plurality of fine holes (eyes) smaller than the size of the object to be measured. As a result, the object to be measured can be reliably collected from the sample stock solution by the filter unit 31. The object to be measured collected by the filter unit 31 is held on the filter unit 31. Moreover, in this Embodiment, since the nucleic acid extracted from the to-be-measured object is passed through the filter part 31, it is good to make the magnitude | size of the hole of the filter part 31 larger than the magnitude | size of a nucleic acid.

As the filter unit 31, a membrane filter made of a material such as cellulose acetate, polyvinylidene fluoride (PVDF), or polyethersulfone (PES) can be used. Further, as the filter unit 31, a capture filter having a function of adsorbing an object to be measured contained in the sample stock solution may be used. By using such a capture filter, it is possible to improve the speed of capturing the object to be measured from the sample stock solution. In this embodiment, the filter unit 31 is, for example, the capture area is planar filter of 20mm ² ~50mm ^2, not limited to this, a filter of other shapes of the cylindrical etc. Also good.

  At least a part of the support portion 32 may be made of a metal material such as aluminum or a high thermal conductive resin. Thereby, the nucleic acid extraction reagent can be heated via the support part 32 by using a heater or the like. In this case, the whole support part 32 may be comprised with the metal material or the high heat conductive resin. Further, when a metal material is used for a part of the support portion 32, the metal material may be embedded in the resin. In the present embodiment, the support portion 32 is made of a metal made entirely of a metal material.

  The retention part 40 is a part where a nucleic acid solution containing a nucleic acid extracted from an object to be measured contained in a sample stock solution is retained, and is provided in the container 10. In the present embodiment, the retention part 40 is a recess provided in the third container part 13. Specifically, the shape of the staying portion 40 is a substantially conical shape, but is not limited thereto, and may be a truncated cone shape, a cylindrical shape, a prismatic shape, or the like. The staying part 40 may have a tapered surface (inclined surface) whose inner diameter gradually decreases.

  Nucleic acid is eluted from the object to be measured captured by the capture unit 30 by the nucleic acid extraction reagent. The solution containing the nucleic acid (nucleic acid solution) passes through the capture unit 30 and is retained in the retention unit 40. That is, the retention part 40 is a storage part for collecting and temporarily storing the nucleic acid solution. The staying part 40 is not provided in the container 10 (third container part 13), but may be a member different from the container 10.

  The heating unit 50 is a member for heating the nucleic acid solution retained in the retention unit 40. In the present embodiment, the heating unit 50 is a part of the container 10. Specifically, the heating unit 50 is the third container unit 13.

  The heating unit 50 may be, for example, a heat transfer member that does not have a heat source and only transfers heat from the outside to the staying unit 40, or a heating device (heater) having a heat source that generates heat from the heating unit 50 itself. Etc. When the heating unit 50 is a heat transfer member, for example, the heating unit 50 receives heat from a heater (not shown) such as a heater block adjacent to the heating unit 50 and conducts the heat to the staying unit 40. Thus, the nucleic acid solution retained in the retention part 40 is heated. On the other hand, when the heating unit 50 itself is a heater such as a heater block, the nucleic acid solution retained in the retention unit 40 is heated by the heat generated by the heating unit 50. The heater block is a heater having a metal block made of a metal such as aluminum or stainless steel.

  The nucleic acid solution staying in the staying part 40 is heated by the heating part 50, whereby the solvent of the nucleic acid solution is evaporated and the nucleic acid solution is concentrated. That is, the concentration of nucleic acid in the nucleic acid solution is increased.

  The first discharge port 61 is a vapor discharge port for discharging the solvent vapor generated when the nucleic acid solution is heated by the heating unit 50 from the container 10. That is, the first discharge port 61 is an exhaust port for exhausting the solvent gas of the nucleic acid solution evaporated from the nucleic acid solution from the container 10 when the nucleic acid solution retained in the retention unit 40 is heated by the heating unit 50. is there.

  The first exhaust port 61 also serves as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed. Therefore, the 1st discharge port 61 is connected to the vacuum pump 101 via piping and a rubber tube.

  Further, the first discharge port 61 also serves as a drain port for discharging the sample stock solution from the container 10. That is, the waste liquid of the sample stock solution injected into the container 10 is discharged from the first discharge port 61. Specifically, the sample stock solution after being injected into the first container unit 11 and passing through the capturing unit 30 is discharged from the first discharge port 61. In the present embodiment, the first discharge port 61 is provided on the side wall of the second container portion 12.

  The first discharge port 61 is a pipe that communicates the outside of the container 10 and the internal space of the second container part 12. Specifically, one end of the first outlet 61 is connected to the second container part 12, and the other end of the first outlet 61 is connected to the nucleic acid via a pipe as shown in FIG. It is connected to a recovery container 500 (specimen waste liquid recovery container) installed in the extraction apparatus 100. In the present embodiment, the first discharge port 61 is formed integrally with the second container portion 12. Note that the first outlet 61 may be a through-hole, like the first inlet 21 and the second inlet 22.

  The sample stock solution discharged from the first discharge port 61 is collected in the collection container 500 through a pipe connected to the first discharge port 61. For example, the waste liquid of the sample stock solution is recovered by being sent from the container 10 to the recovery container 500 by suction by the vacuum pump 101.

  The vacuum pump 101 is an example of a pressure adjusting unit for adjusting the pressure in the container 10. In the present embodiment, the piping of the vacuum pump 101 is connected to the second container portion 12. The pressure in the container 10 can be adjusted by a leak valve 103 provided in the piping of the vacuum pump 101 as shown in FIG. That is, the degree of pressure reduction by the vacuum pump 101 can be adjusted by the leak valve 103.

  In the present embodiment, the piping of the vacuum pump 101 is connected to the first discharge port 61. That is, the decompression port (decompression suction port) of the vacuum pump 101 also serves as the first discharge port 61 provided in the second container portion 12.

[Nucleic acid extraction method]
Next, a nucleic acid extraction method using the nucleic acid extraction apparatus 100 will be described with reference to FIGS. 3A to 3H with reference to FIG. 3A to 3H are schematic cross-sectional views for explaining the nucleic acid extraction method according to the first embodiment.

  First, as shown in FIG. 3A, the sample stock solution 210 is injected into the nucleic acid extraction unit 1. Specifically, the sample input cup 200 containing the sample stock solution 210 including the object 220 to be measured is placed at a predetermined position of the nucleic acid extraction apparatus 100 (see FIG. 1), and the sample stock solution is transferred from the sample input cup 200 to the container 10. 210 is inserted. For example, 100 ml of the sample stock solution 210 is introduced.

  The sample stock solution 210 of the sample insertion cup 200 is injected into the first container unit 11 through the first injection port 21, passes through the capturing unit 30, and flows into the second container unit 12. At this time, when the sample stock solution 210 passes through the capturing unit 30, the measurement target 220 contained in the sample stock solution 210 is captured by the capturing unit 30. In the present embodiment, the measurement object 220 contained in the sample stock solution 210 is captured by the filter unit 31 and remains on the filter unit 31 when passing through the filter unit 31 of the capturing unit 30.

  As shown in FIG. 3A, the specimen stock solution 210 that has passed through the capture unit 30 is discharged from the container 10 (second container unit 12) through the first discharge port 61 as waste liquid, and the nucleic acid extraction apparatus 100 (see FIG. 1). Are collected in a collection container 500 installed at a predetermined position.

  At this time, in the present embodiment, the object to be measured 220 included in the sample stock solution 210 is captured by the capturing unit 30 by suction filtration. Specifically, the inside of the container 10 is decompressed by exhausting (suctioning) the air in the container 10 from the first discharge port 61 by the vacuum pump 101, and the specimen stock solution 210 injected into the container 10 is sucked in. Thus, the object to be measured 220 is captured by the capturing unit 30 from the sample stock solution 210 by filtration. As a result, the sample stock solution 210 injected into the container 10 can be quickly passed through the capturing unit 30, and the waste solution of the sample stock solution 210 can be quickly discharged from the container 10.

  When injecting the sample stock solution 210 into the container 10, the sample stock solution 210 may be further pressurized from above. For example, the space region above the sample stock solution 210 in the sample insertion cup 200 may be pressurized. Thereby, the sample stock solution 210 can be discharged from the container 10 more efficiently.

  In this way, by injecting and discharging the sample stock solution 210 into the container 10, as shown in FIG. 3B, the object to be measured 220 in the sample stock solution 210 remains on the filter unit 31 of the capturing unit 30.

  Next, the nucleic acid extraction reagent 310 is injected into the container 10, and the nucleic acid 230 is extracted from the measurement object 220 captured by the capturing unit 30. The amount of the nucleic acid extraction reagent 310 to be injected is, for example, 50 μl to 200 μl, and is 100 μl in this embodiment.

  Specifically, as shown in FIG. 3C, the nucleic acid extraction reagent 310 is injected into the container 10 from the nucleic acid extraction reagent container 300 installed at a predetermined position of the nucleic acid extraction apparatus 100 (see FIG. 1). The nucleic acid extraction reagent 310 of the nucleic acid extraction reagent container 300 is injected into the first container part 11 through the second injection port 22. Specifically, nucleic acid extraction is performed from the nucleic acid extraction reagent container 300 into the first container portion 11 by passing the injection needle of the nucleic acid extraction reagent container 300 through a rubber stopper 22a provided in the second injection port 22 (through hole). Reagent 310 is injected. In addition, before injecting the nucleic acid extraction reagent 310 into the container 10, a dead bacteria inactive treatment reagent or the like may be injected into the container 10.

  When the nucleic acid extraction reagent 310 is injected into the container 10, the measurement object 220 captured by the capturing unit 30 reacts with the nucleic acid extraction reagent 310. Thereby, the nucleic acid 230 can be extracted from the DUT 220. That is, a nucleic acid extraction reaction in which the nucleic acid 230 is extracted from the object to be measured 220 is performed. Specifically, as shown in FIG. 3D, the nucleic acid 230 is eluted from the DUT 220 to the nucleic acid extraction reagent 310, and a nucleic acid solution 320 containing the nucleic acid 230 is generated. Thereby, the cell membrane of the object to be measured 220 (for example, a microorganism) and the nucleic acid 230 are separated.

  At this time, as shown in FIG. 3D, the nucleic acid extraction reaction of the DUT 220 may be performed by holding the nucleic acid extraction reagent 310 injected into the container 10 on the capture unit 30. As a result, the object 220 to be measured is immersed in the nucleic acid extraction reagent 310 for a certain time, and the nucleic acid 230 can be easily eluted from the object 220 to be measured. As a result, the nucleic acid 230 can be extracted (eluted) efficiently.

  When the nucleic acid extraction reagent 310 is held on the capturing unit 30, the inside of the second container unit 12 is pressurized by the vacuum pump 101 so that the pressure in the second container unit 12 is higher than the pressure in the first container unit 11. That's fine. Thereby, the nucleic acid extraction reagent 310 injected into the container 10 can be held on the capturing unit 30.

  Thereafter, after the injection of the nucleic acid extraction reagent 310 is completed and the nucleic acid extraction reaction of the object 220 to be measured on the capturing unit 30 is completed, the nucleic acid solution 320 containing the extracted nucleic acid 230 is passed through the capturing unit 30 to the staying unit 40. Let it stay.

  Specifically, by stopping pressurization in the second container part 12 by the vacuum pump 101, the nucleic acid solution 320 staying on the capture part 30 is transferred to the second container part 12 as shown in FIG. 3E. Can be moved. For example, by opening the leak valve 103, the inside of the second container part 12 is brought to normal pressure, or the pressure in the second container part 12 is adjusted so that the degree of decompression is lower than the degree of decompression during suction filtration. do it.

  Note that the method for holding the nucleic acid extraction reagent 310 on the capturing unit 30 is not limited to the method for adjusting the pressure in the second container unit 12 as described above. For example, the filter unit 31 of the capturing unit 30 can be realized by using a filter that holds the nucleic acid extraction reagent 310 on the filter unit 31 when the inside of the second container unit 12 is not decompressed.

  In this case, although not shown, after the injection of the nucleic acid extraction reagent 310 is completed and the nucleic acid extraction reaction of the object 220 on the capture unit 30 is completed by the nucleic acid extraction reagent 310 held on the capture unit 30, The pressure inside the second container part 12 may be reduced by the vacuum pump 101 so that the pressure inside the first container part 11 is higher than the pressure inside the second container part 12. As a result, the nucleic acid solution 320 remaining on the capture unit 30 after completion of the nucleic acid extraction reaction can be moved to the second container unit 12. That is, the nucleic acid solution 320 can be moved to the second container part 12 using the pressure difference.

  As shown in FIG. 3F, the nucleic acid solution 320 that has passed through the capture unit 30 and moved to the second container unit 12 stays in the retention unit 40 that is a recess of the third container unit 13. That is, the nucleic acid solution 320 is temporarily stored in the retention part 40.

  In this way, the nucleic acid solution 320 containing the nucleic acid 230 extracted by the capture unit 30 is retained in the retention unit 40 through the capture unit 30.

  Next, as shown in FIG. 3G, the nucleic acid solution 320 staying in the staying portion 40 is heated under reduced pressure, and the solvent vapor of the nucleic acid solution 320 is discharged from the first discharge port 61 to the outside of the container 10. Concentrate solution 320. That is, by heating the nucleic acid solution 320 retained in the retention part 40 while reducing the pressure, the solvent component of the nucleic acid solution 320 is vaporized (evaporated), and the vaporized vapor of the solvent component is discharged from the first outlet 61 to the container 10. It can be quickly exhausted to the outside. Thereby, the nucleic acid solution 320 can be efficiently concentrated in a short time. That is, a highly concentrated nucleic acid solution 320 having a high concentration of the nucleic acid 230 can be obtained in a short time.

  Specifically, the temperature around the staying part 40 is heated to, for example, 90 ° C. by the heating part 50, and at the same time, the inside of the container 10 is decompressed by exhausting the air in the container 10 from the first discharge port 61 by the vacuum pump 101. The pressure inside the container 10 is reduced. The reduced pressure state is, for example, 0.01 MPa to 0.001 MPa. In the present embodiment, 100 μl of the nucleic acid solution 320 is concentrated to 10 μl. That is, the nucleic acid solution 320 is concentrated so as to be 10% of the amount before heating. Note that the amount of the concentrated nucleic acid solution 320 is not limited to 10 μl.

  In the present embodiment, since the heating unit 50 is a metal member, the heating unit 50 is heated using the heater 70. Thereby, the nucleic acid solution 320 is heated by the heat of the heater 70 conducted to the heating unit 50. The heater 70 may be provided in the nucleic acid extraction unit 1 or may be provided in the nucleic acid extraction apparatus 100. In addition, when the heating part 50 itself has a heater function, the heater 70 is unnecessary.

  Next, as shown in FIG. 3H, the nucleic acid solution 320 containing the nucleic acid 230 is recovered. Specifically, the nucleic acid solution 320 in the container 10 is recovered in the recovery container 400 by removing the pipe from the first outlet 61 and tilting the nucleic acid extraction unit 1.

  Thereafter, although not shown, a nucleic acid test reagent is mixed into the concentrated nucleic acid solution 320 to obtain a test solution. For example, 10 μl of nucleic acid solution 320 is mixed with 90 μl of nucleic acid test reagent to make 100 μl of test solution. The amount of the nucleic acid test reagent is not limited to 90 μm, and may be, for example, about 50 μl to 150 μl. Further, the amount of the nucleic acid solution 320 after mixing the nucleic acid test reagent is not limited to 100 μl, and may be, for example, 50 μl to 150 μl. The nucleic acid test reagent (PCR reagent) includes, for example, a nucleic acid staining fluorescent reagent for testing the amount of nucleic acid with fluorescence emission intensity, a reaction reagent for amplifying nucleic acid, and the like. The reaction reagent is, for example, a PCR primer, a polymerase enzyme, a buffer or the like.

  Note that the mixed solution of the nucleic acid solution 320 and the nucleic acid test reagent is amplified by nucleic acid by flow PCR using a test chip (PCR chip or the like) in which a micro flow path as a test flow path is formed, and amplified by an optical detection device. The measured object can be detected by measuring the nucleic acid.

[Summary]
As described above, the nucleic acid extraction apparatus 100 according to the present embodiment captures the object to be measured 220 contained in the container 10, the first injection port 21, and the sample stock solution 210 injected from the first injection port 21 by filtration. Capture unit 30, second injection port 22 for injecting nucleic acid extraction reagent 310 into container 10, retention unit 40 in which nucleic acid solution 320 containing nucleic acid 230 extracted from object 220 is retained, and retention A heating unit 50 for heating the nucleic acid solution 320 retained in the unit 40, and a first outlet 61 for discharging the vapor of the solvent generated when the nucleic acid solution 320 is heated by the heating unit 50 from the container 10. I have. The first discharge port 61 also serves as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed.

  As described above, according to the nucleic acid extraction apparatus 100 in the present embodiment, the nucleic acid solution 320 retained in the retention unit 40 can be heated and concentrated, so that a highly concentrated nucleic acid solution 320 can be obtained. Therefore, it is possible to realize a nucleic acid extraction apparatus that can inspect the object 220 with high sensitivity and high accuracy without using a large amount of nucleic acid inspection reagent.

  Moreover, the first discharge port 61 that discharges the steam generated when the nucleic acid solution 320 is heated also serves as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed. Specifically, an exhaust line for creating a depressurized state for efficiently discharging the vapor of the solvent generated when the nucleic acid solution 320 is heated, and a depressurized state for sucking and filtering the object to be measured 220 to the capturing unit 30 Can be used in common with the exhaust line. Thereby, simplification and size reduction of the nucleic acid extraction apparatus 100 can be achieved.

  Further, in the nucleic acid extraction method according to the present embodiment, the sample stock solution injected into the container 10 by depressurizing the inside of the container 10 by exhausting the air in the container 10 from the first outlet 61 provided in the container 10. The sample to be measured 220 is captured by the capture unit 30 by filtration from the sample stock solution 210 by aspirating 210, and the nucleic acid 230 is extracted from the sample 220 captured by the capture unit 30 by injecting the nucleic acid extraction reagent 310 into the container 10. Then, the nucleic acid solution 320 containing the extracted nucleic acid 230 is retained in the retention unit 40 through the capture unit 30, the nucleic acid solution 320 retained in the retention unit 40 is heated under reduced pressure, and the solvent vapor of the nucleic acid solution 320 is changed to a first level. The nucleic acid solution 320 is concentrated by being discharged from the outlet 61 to the outside of the container 10.

  As described above, according to the nucleic acid extraction method of the present embodiment, the nucleic acid solution 320 containing the nucleic acid 230 extracted from the measurement target 220 is heated and concentrated. Thereby, a high concentration nucleic acid solution 320 can be obtained. Therefore, the test object 220 can be tested with high sensitivity and high accuracy without using a large amount of nucleic acid test reagent.

  In addition, the first discharge port 61 that discharges the steam generated when the nucleic acid solution 320 is heated is used as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed. Thereby, nucleic acid extraction can be performed using the nucleic acid extraction apparatus 100 simplified and miniaturized.

(Modification 1 of Embodiment 1)
Next, a nucleic acid extraction apparatus according to Modification 1 of Embodiment 1 will be described with reference to FIG. FIG. 4 is a cross-sectional view of a nucleic acid extraction unit 1A used in the nucleic acid extraction apparatus according to Modification 1 of Embodiment 1. The configuration other than the nucleic acid extraction unit 1A is the same as that in the first embodiment.

  As shown in FIG. 4, in the nucleic acid extraction unit 1 </ b> A according to this modification, the heating unit 50 includes a third container unit 13 made of a heat transfer member such as aluminum and a heater 70.

  The heater 70 is, for example, a heater block, and is thermally connected to the staying part 40 (third container part 13) and thermally connected to the metal support part 32 of the capturing part 30. Specifically, the heater 70 is provided so as to straddle the third container portion 13 and the support portion 32 while being in contact with each of the third container portion 13 and the support portion 32.

  Thereby, the stay part 40 and the capture part 30 can be heated by the heater 70. Here, when the nucleic acid extraction reagent 310 is injected and the nucleic acid 230 is extracted from the measurement object 220, for example, the nucleic acid extraction reagent 310 may be heated to about 70 ° C. in order to promote the nucleic acid extraction reaction. As in the example, the nucleic acid extraction reagent 310 held on the capture unit 30 by the heater 70 can be heated to 70 ° C. by thermally connecting the heater 70 to the metal support portion 32.

  Thus, in this modification, not only the nucleic acid solution 320 retained in the retention unit 40 by the heater 70 is heated, but also the nucleic acid extraction reagent 310 on the capture unit 30 is heated. Thereby, since the capture unit 30 (nucleic acid extraction reagent 310) and the retention unit 40 (nucleic acid solution 320) can be heated by sharing the heater 70, the nucleic acid extraction apparatus can be further simplified and miniaturized.

(Modification 2 of Embodiment 1)
Next, a nucleic acid extraction apparatus according to Modification 2 of Embodiment 1 will be described with reference to FIG. FIG. 5 is a cross-sectional view of the nucleic acid extraction unit 1B used in the nucleic acid extraction apparatus according to the second modification of the first embodiment. The configuration other than the nucleic acid extraction unit 1B is the same as that in the first embodiment.

  As shown in FIG. 5, in the nucleic acid extraction unit 1B according to this modification, the heating unit 50B includes a third container unit 13B of the container 10B, a support unit 32B of the capturing unit 30B, and a heater 70.

  The third container portion 13B and the support portion 32B are heat transfer members made of a metal material such as aluminum. Moreover, the 3rd container part 13B and the support part 32B are integrated via the connection part 51B.

  The configuration of the heater 70 is the same as that of the first modification, but in this modification, the heater 70 is disposed adjacent to the connecting portion 51B. Specifically, the heater 70 is in contact with the connecting portion 51B.

  Thus, in the present modification, the connecting portion 51B that connects the third container portion 13B and the support portion 32B is adjacent to the heater 70. Thereby, the heat generated by the heater 70 can be efficiently conducted to the third container portion 13B and the support portion 32B via the connecting portion 51B. Therefore, it is possible to efficiently heat the capture unit 30B (nucleic acid extraction reagent 310) and the retention unit 40 (nucleic acid solution 320).

  Further, in this modification, as in Modification 1, since the heater 70 is shared by the heating of the capturing unit 30B and the heating of the staying unit 40, the nucleic acid extraction apparatus can be further simplified and miniaturized.

  Further, in the present modification, the third container portion 13B and the support portion 32B are connected by the connecting portion 51B. Therefore, if one of the third container portion 13B and the support portion 32B is heated, the other is heated. It can be heated by conduction. Therefore, since the nucleic acid extraction reagent 310 and the nucleic acid solution 320 can be heated by one heater 70, the nucleic acid extraction apparatus can be simplified.

(Modification 3 of Embodiment 1)
Next, a nucleic acid extraction apparatus according to Modification 3 of Embodiment 1 will be described with reference to FIG. FIG. 6 is a cross-sectional view of a nucleic acid extraction unit 1C used in the nucleic acid extraction apparatus according to the third modification of the first embodiment. The configuration other than the nucleic acid extraction unit 1C is the same as that of the first embodiment.

  As shown in FIG. 6, in the nucleic acid extraction unit 1C in the present modification, as in the modification 2, the heating unit 50C includes the third container unit 13C of the container 10C, the support unit 32C of the capturing unit 30C, and the heater. 70C.

  Also in this modification, the 3rd container part 13C and the support part 32C are heat-transfer members which consist of metal materials, such as aluminum, for example, and the 3rd container part 13C and the support part 32C are integrated via the connection part 51C. Has been.

  The heater 70C is adjacent to the connecting portion 51C as in the second modification. Specifically, the heater 70C and the connecting portion 51C are brought into contact with each other.

  On the other hand, regarding the configuration of the heater 70C, unlike the second modification, the heater 70C is an induction heater having a magnetic body and a coil around which the magnetic body is wound. That is, the heater 70C generates heat by electromagnetic induction.

  Thus, in the present modification, as in Modification 2, the connecting portion 51C that connects the third container portion 13C and the support portion 32C and the heater 70C are adjacent to each other. Thereby, the heat generated by the heater 70C can be efficiently conducted to the third container portion 13C and the support portion 32C via the connecting portion 51C.

  Also in this modified example, similarly to the modified example 2, since the heater 70C is shared by the heating of the capturing unit 30C and the heating of the staying unit 40, the nucleic acid extraction apparatus can be further simplified and miniaturized.

  Moreover, in this modification, since the 3rd container part 13C and the support part 32C are connected by the connection part 51C, if either the 3rd container part 13C or the support part 32C is heated, the other will be the heat. The nucleic acid extraction reagent 310 and the nucleic acid solution 320 can be heated by one heater 70.

  Moreover, in this modification, since the induction heater is used as the heater 70C, the capturing unit 30C and the staying unit 40 can be efficiently heated.

  As in the case of the nucleic acid extraction unit 1D shown in FIG. 7, a heater 70C (induction heater) may be used instead of the heater 70 in the nucleic acid extraction unit 1 shown in FIG. 3G.

(Modification 4 of Embodiment 1)
Next, a nucleic acid extraction apparatus according to Modification 4 of Embodiment 1 will be described with reference to FIG. FIG. 8 is a cross-sectional view of a nucleic acid extraction unit 1E used in the nucleic acid extraction apparatus according to the fourth modification of the first embodiment. The configuration other than the nucleic acid extraction unit 1E is the same as that in the first embodiment.

  As shown in FIG. 8, in the nucleic acid extraction unit 1 </ b> E according to this modification, the support part 32 </ b> E of the capturing part 30 </ b> E has an exposed part 32 </ b> Ea formed along the outer surface of the container 10. The support portion 32E has an L-shaped cross section and is made of a metal material such as aluminum.

  Moreover, the metal member 33E is provided in the eye plate part of the support part 32E. In the present embodiment, the metal member 33E is integrated with the support portion 32E. The metal member 33 is, for example, a metal piece (metal plate) or a metal column. A part of the metal member 33E is immersed in the nucleic acid solution 320 retained in the retention part 40. That is, the metal member 33 </ b> E is shaped so as to come into contact with the nucleic acid solution 320 when the nucleic acid solution 320 is retained in the retention part 40. For example, the metal member 33E is formed in a long shape so as to extend from the support portion 32E toward the staying portion 40.

  The heating unit 50E is a heater (heater) having a heat source such as a heater block. The heating part 50E is disposed adjacent to the exposed part 32Ea of the support part 32E. Specifically, the heating unit 50E and the support unit 32E are brought into contact with each other.

  Thus, in this modification, the heat generated in the heating unit 50E is conducted to the support unit 32E through the exposed portion 32Ea. The heat conducted to the support portion 32E is conducted to the nucleic acid solution 320 retained in the retention portion 40 via the metal member 33E. Thereby, the nucleic acid solution 320 can be heated. That is, the nucleic acid solution 320 staying in the staying part 40 is heated by the heat of the heating part 50E conducted through the metal member 33E.

  As described above, according to this modification, the metal member 33E is brought into contact with the nucleic acid solution 320 retained in the retention unit 40, and the heat generated in the heating unit 50E is conducted to the nucleic acid solution 320 through the metal member 33E. The nucleic acid solution 320 staying in the staying section 40 is heated and concentrated.

  Thereby, since the nucleic acid solution 320 staying in the staying part 40 can be directly heated by the metal member 33E, the nucleic acid solution 320 can be concentrated in a short time. In addition, both the capture unit 30E (nucleic acid extraction reagent 310) and the retention unit 40 (nucleic acid solution 320) can be heated with a simple structure.

  As in the case of the nucleic acid extraction unit 1F shown in FIG. 9, a spiral metal member 33F may be used instead of the metal member 33E in the nucleic acid extraction unit 1E shown in FIG. In this case, as shown in FIG. 10, a spiral through hole (slit) is formed in the support portion 32F of the capturing portion 30F. The metal member 33F is a spiral piece cut out spirally from the support part 32F of the capturing part 30F and extended downward.

  In the present modification, the third container part 13 may not be a heating part. Therefore, in FIGS. 8 and 9, the material of the third container part 13 is the same as the material of the second container part 12. It may be different or different. Specifically, the material of the third container portion 13 may be made of metal, resin, or the like. In addition, you may make the 3rd container part 13 function as a heating part.

(Embodiment 2)
[Nucleic acid extraction equipment]
Next, the configuration of the nucleic acid extraction apparatus 100A and the nucleic acid extraction unit 2 according to Embodiment 2 will be described with reference to FIGS. FIG. 11 is a perspective view schematically showing a nucleic acid extraction apparatus 100A according to the second embodiment. FIG. 12 is a cross-sectional view of the nucleic acid extraction unit 2 according to the second embodiment.

  As shown in FIG. 11, the nucleic acid extraction apparatus 100A is an apparatus for extracting nucleic acid from an object to be measured contained in a sample stock solution, like the nucleic acid extraction apparatus 100 in the first embodiment. The vacuum pump 101 and the reaction box 102 are provided. The nucleic acid extraction apparatus 100 in the present embodiment further includes a liquid feed pump 104. The liquid feed pump 104 sends the nucleic acid solution 320 concentrated by the nucleic acid extraction unit 2 to the collection container 400.

  As shown in FIG. 12, the nucleic acid extraction unit 2 is similar to the nucleic acid extraction unit 1 in the first embodiment. The container 10, the first injection port 21, the second injection port 22, the capture unit 30, and the retention Unit 40, heating unit 50, and first discharge port 61. The nucleic acid extraction unit 2 in the present embodiment further includes a second outlet 62.

  The second outlet 62 is a nucleic acid solution outlet for discharging the nucleic acid solution containing the nucleic acid extracted from the measurement object from the container 10. In the present embodiment, the second discharge port 62 is provided on the lower wall of the second container portion 12.

  The second discharge port 62 is a pipe that communicates the outside of the container 10 and the internal space of the container 10. Specifically, one end of the second outlet 62 is connected to the third container part 13, and the other end of the second outlet 62 is connected to the nucleic acid extraction apparatus 100 via a pipe or a rubber tube. It is connected to the installed collection container 400. In the present embodiment, the second discharge port 62 is formed integrally with the third container portion 13.

  The nucleic acid solution discharged from the second outlet 62 is recovered in the recovery container 400 through a pipe connected to the second outlet 62. For example, the nucleic acid solution retained in the retention unit 40 is collected by being fed from the container 10 to the collection container 400 by the liquid feeding pump 104.

  A valve 105 is provided in the pipe connected to the second outlet 62. The pipe can be opened or closed by the valve 105. The valve 105 is provided, for example, in the liquid feed pump 104 in FIG. 11, but is not limited thereto.

[Nucleic acid extraction method]
Next, a nucleic acid extraction method using the nucleic acid extraction apparatus 100A will be described using FIGS. 13A to 13H with reference to FIG. 13A to 13H are schematic cross-sectional views for explaining the nucleic acid extraction method according to the second embodiment.

  First, as shown in FIG. 13A, the sample stock solution 210 is injected into the nucleic acid extraction unit 1 in the same manner as in the first embodiment. At this time, the valve 105 provided in the pipe connected to the second discharge port 62 is closed.

  The sample stock solution 210 of the sample insertion cup 200 is injected into the first container unit 11 through the first injection port 21, passes through the capturing unit 30, and flows into the second container unit 12. At this time, when the sample stock solution 210 passes through the capturing unit 30, the measurement target 220 contained in the sample stock solution 210 is captured by the capturing unit 30. Also in this embodiment, the sample stock solution 210 is filtered by being sucked by the vacuum pump 101.

  In this way, when the sample stock solution 210 is injected into the container 10 and discharged, the object to be measured 220 accumulates on the filter unit 31 of the capturing unit 30 as shown in FIG. 13B.

  Next, as shown in FIG. 13C, the nucleic acid 230 is extracted from the object to be measured 220 captured by the capturing unit 30 by injecting the nucleic acid extraction reagent 310 into the container 10 as in the first embodiment. At this time, the valve 105 remains closed.

  When the nucleic acid extraction reagent 310 is injected into the container 10, the measurement object 220 captured by the capture unit 30 reacts with the nucleic acid extraction reagent 310, and as shown in FIG. As a result, the nucleic acid solution 320 containing the nucleic acid 230 is generated. Also in the present embodiment, the nucleic acid extraction reaction of the object to be measured 220 is performed by holding the nucleic acid extraction reagent 310 injected into the container 10 on the capturing unit 30.

  Thereafter, after the injection of the nucleic acid extraction reagent 310 is completed and the nucleic acid extraction reaction of the measurement object 220 on the capture unit 30 is completed, the extracted nucleic acid is performed in the same manner as in Embodiment 1 as shown in FIG. 13E. When the nucleic acid solution 320 containing 230 passes through the capture unit 30, the nucleic acid solution stays in the retention unit 40 as shown in FIG. 3F. At this time, the valve 105 remains closed.

  Next, as shown in FIG. 13G, in the same manner as in the first embodiment, the nucleic acid solution 320 staying in the staying section 40 is heated under reduced pressure, and the solvent vapor of the nucleic acid solution 320 is discharged from the first outlet 61. The nucleic acid solution 320 is concentrated by discharging to the outside of the container 10. At this time, the valve 105 remains closed.

  13G does not show the heater, the heater 70 may be used as in FIG. 3G, or when the heating unit 50 itself has a heater function, the heat generated by the heating unit 50 may be used. The nucleic acid solution 320 may be heated.

  Next, as shown in FIG. 13H, the nucleic acid solution 320 containing the nucleic acid 230 is recovered. In this case, unlike Embodiment 1, the nucleic acid solution 320 is recovered from the second outlet 62 instead of the first outlet 61.

  Specifically, the concentrated nucleic acid solution 320 is discharged from the container 10 through the second discharge port 62 by opening the valve 105 of the pipe connected to the second discharge port 62, and the discharged nucleic acid solution 320 is discharged. Is recovered in a recovery container 400 installed at a predetermined position of the nucleic acid extraction apparatus 100A (see FIG. 11). In this case, the nucleic acid solution 320 is fed to the collection container 400 by the liquid feed pump 104.

  Thereafter, although not shown in the drawings, the object to be measured 220 can be inspected by mixing a nucleic acid test reagent with the nucleic acid solution 320 and amplifying the nucleic acid using a test chip or the like, as in the first embodiment.

[Summary]
As described above, the nucleic acid extraction apparatus 100A according to the present embodiment is similar to the nucleic acid extraction apparatus 100 according to the first embodiment. The container 10, the first injection port 21, the capturing unit 30, the second injection port 22, and the like. The retention part 40, the heating part 50, and the 1st discharge port 61 are provided. The first discharge port 61 also serves as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed.

  Thereby, the same effects as those of the first embodiment can be obtained. That is, it is possible to realize a nucleic acid extraction apparatus that can inspect the object 220 with high sensitivity and high accuracy without using a large amount of nucleic acid inspection reagent. In addition, the nucleic acid extraction apparatus 100 can be simplified and downsized.

  Furthermore, the nucleic acid extraction apparatus 100A according to the present embodiment includes a second discharge port 62 for discharging the nucleic acid solution 320 from the container 10. That is, the first discharge port 61 for discharging the waste liquid of the sample stock solution 210 or the air in the container 10 and the second discharge port 62 for discharging the nucleic acid solution 320 are separated.

  Thereby, in Embodiment 1, since the 1st discharge port 61 and the 2nd discharge port 62 were shared, when the nucleic acid solution 320 is discharged | emitted from the container 10, the nucleic acid extraction unit 1 is inclined or the Although a rotation mechanism for tilting is required, in this embodiment, since the nucleic acid extraction unit 1 is not tilted or a rotation mechanism is not required, it is possible to avoid complication of the apparatus configuration, and it is simple. A nucleic acid extraction apparatus having a structure can be realized.

  As shown in FIG. 14, the nucleic acid extraction apparatus 100 </ b> A according to the present embodiment may further include a pump unit 106 for introducing bubbles 106 a into the nucleic acid solution 320 retained in the retention unit 40.

  In this case, when the nucleic acid solution 320 retained in the retention unit 40 is heated by the heating unit 50, the bubbles 106 a are introduced into the nucleic acid solution 320 by the pump unit 106.

  Thereby, since the solvent of the nucleic acid solution 320 can be efficiently vaporized, the nucleic acid solution 320 can be concentrated in a short time.

  Moreover, the bubble 106a introduced into the nucleic acid solution 320 may be hot air. The temperature of the hot air is 90 ° C., for example.

  In this way, by introducing hot air as the bubble 106a into the nucleic acid solution 320, the solvent of the nucleic acid solution 320 can be vaporized more efficiently.

(Embodiment 3)
Next, the configuration of the nucleic acid extraction apparatus and the nucleic acid extraction unit 3 according to Embodiment 3 will be described with reference to FIGS. 15 and 16. 15 and 16 are cross-sectional views of the nucleic acid extraction unit 3 used in the nucleic acid extraction apparatus according to the third embodiment. FIG. 15 shows a state when the elastic member 81 is elastically deformed, and FIG. 16 shows a state when the elastic member 81 is not elastically deformed. The configuration other than the nucleic acid extraction unit 3 is the same as in the first and second embodiments.

  As shown in FIGS. 15 and 16, the nucleic acid extraction unit 3 in the present embodiment is similar to the nucleic acid extraction unit 2 in the second embodiment, in the container 10, the first inlet 21, and the second inlet 22. And a capture unit 30, a staying unit 40, a first discharge port 61, and a second discharge port 62.

  The nucleic acid extraction unit 3 in the present embodiment further includes an opening / closing mechanism 80 for opening or closing the second outlet 62.

  The opening / closing mechanism 80 includes a film-like elastic member 81 that opens or closes the second discharge port 62 by elastic deformation, and a movable member 82 that elastically deforms the elastic member 81.

  The elastic member 81 is made of an elastically deformable material. The elastic member 81 is made of, for example, a resin material having elasticity, such as an elastomer. The elastic member 81 is elastically deformed when a pressure is applied, and returns to its original shape by an elastic restoring force when the pressure is not applied. In the present embodiment, the elastic member 81 is a flat thin plate-like rectangular rubber film (elastic film), and is made of, for example, silicone rubber. The elastic member 81 preferably has a high thermal conductivity, and a high thermal conductive silicone rubber may be used.

  The movable member 82 is, for example, a solenoid, and moves in a direction approaching the elastic member 81 or moves in a direction away from the elastic member 81. The movable member 82 is made of, for example, a metal material. In the present embodiment, the movable member 82 has a heater 70. That is, the movable member 82 has a heater function. The heat generated by the movable member 82 (heater 70) can be conducted to the staying part 40 via the elastic member 81.

  As shown in FIG. 15, the nucleic acid solution 320 can be retained in the retention part 40 by closing the second discharge port 62 with the elastic member 81. Specifically, the second discharge port 62 is closed by the elastic member 81 by moving the movable member 82 upward and applying pressure to the elastic member 81 to elastically deform the elastic member 81. That is, the second discharge port 62 is closed by pressing the elastically deformed elastic member 81 against the second discharge port 62.

  Then, with the second discharge port 62 closed, the nucleic acid solution 320 retained in the retention unit 40 is heated by the movable member 82 (heater 70). That is, since the movable member 82 is in contact with the elastic member 81 in a state where the second discharge port 62 is closed, the heat generated by the movable member 82 (heater 70) is retained through the elastic member 81. Conducted to the nucleic acid solution 320 retained in 40. Thereby, the nucleic acid solution 320 can be heated and concentrated.

  On the other hand, as shown in FIG. 16, the concentrated nucleic acid solution 320 can be discharged from the container 10 by opening the second discharge port 62 by the elastic member 81. Specifically, the second discharge port 62 is opened by moving the movable member 82 downward to return the elastic member 81 to the original flat state. That is, the second discharge port 62 is opened by separating the movable member 82 from the elastic member 81. As a result, the nucleic acid solution 320 in the retention part 40 is discharged from the container 10 through the through hole 81 a provided in the elastic member 81.

  As described above, according to the nucleic acid extraction device in the present embodiment, as in the nucleic acid extraction device 100 in the first embodiment, the container 10, the first injection port 21, the capture unit 30, the second injection port 22, The retention part 40, the heating part 50, and the 1st discharge port 61 are provided. The first discharge port 61 also serves as an exhaust port for exhausting the air in the container 10 when the inside of the container 10 is decompressed.

  Thereby, the same effects as those of the first embodiment can be obtained. That is, it is possible to realize a nucleic acid extraction apparatus that can inspect the object 220 with high sensitivity and high accuracy without using a large amount of nucleic acid inspection reagent. In addition, the nucleic acid extraction apparatus 100 can be simplified and downsized.

  In this embodiment, since the movable member 82 for elastically deforming the elastic member 81 for opening and closing the second discharge port 62 includes the heater 70, the apparatus configuration can be further simplified. it can.

  Furthermore, since the movable member 82 (heater 70) is brought close to the nucleic acid solution 320 via the film-like elastic member 81, the nucleic acid solution 320 can be efficiently heated. Therefore, the nucleic acid solution 320 can be concentrated efficiently.

(Modification 1 of Embodiment 3)
Next, a nucleic acid extraction apparatus according to Modification 1 of Embodiment 3 will be described with reference to FIGS. 17 and 18 are cross-sectional views of the nucleic acid extraction unit 4 used in the nucleic acid extraction apparatus according to the first modification of the third embodiment. FIG. 17 shows a state when the elastic member 81A is elastically deformed, and FIG. 18 shows a state when the elastic member 81A is not elastically deformed. The configuration other than the nucleic acid extraction unit 4 is the same as that of the third embodiment.

  As shown in FIGS. 17 and 18, the nucleic acid extraction unit 4 in the present modification is similar to the nucleic acid extraction unit 3 in the third embodiment in that the container 10, the first injection port 21, the second injection port 22, The capturing unit 30, the staying unit 40, the first discharge port 61, the second discharge port 62, and the opening / closing mechanism 80 </ b> A for opening or closing the second discharge port 62.

  The opening / closing mechanism 80A includes a film-like elastic member 81A that opens or closes the second discharge port 62 by elastic deformation, and a movable member 82 that elastically deforms the elastic member 81.

  The elastic member 81A is made of an elastically deformable material, like the elastic member 81 in the third embodiment. The elastic member 81A is elastically deformed when a pressure is applied, and is restored by an elastic restoring force when the pressure is no longer applied. To return to the original shape.

  Also in this modified example, the elastic member 81A is a flat thin plate-like rectangular rubber film (elastic film), but the elastic member 81A in the modified example functions as a heating unit. The elastic member 81A is a heater that generates heat by the elastic member 81A itself, and is a film heater such as a silicone rubber heater.

  Then, as shown in FIG. 17, the nucleic acid solution 320 can be retained in the retention part 40 by closing the second discharge port 62 with the elastic member 81A. Specifically, as in the third embodiment, the second discharge port 62 is closed by the elastic member 81A by moving the movable member 82 upward and applying pressure to the elastic member 81A to cause elastic deformation. .

  In the present modification, the nucleic acid solution 320 retained in the retention part 40 is heated by the elastic member 81A generating heat while the second discharge port 62 is closed. Thereby, the nucleic acid solution 320 can be directly heated and concentrated by the elastic member 81A.

  On the other hand, as shown in FIG. 18, the concentrated nucleic acid solution 320 can be discharged from the container 10 by opening the second discharge port 62 with the elastic member 81A. Specifically, as in the third embodiment, the second discharge port 62 is opened by moving the movable member 82 downward to return the elastic member 81A to the original flat state. Thereby, the nucleic acid solution 320 of the retention part 40 is discharged | emitted from the container 10 through the through-hole 81a provided in 81 A of elastic members.

  As described above, according to the nucleic acid extraction apparatus in the present modification, the nucleic acid solution 320 can be heated by the elastic member 81A that directly touches the nucleic acid solution 320, so that the nucleic acid solution 320 can be heated very efficiently. In addition, since the elastic member 81A serves as the opening / closing member serving as the opening / closing mechanism 80A and the heating unit, the configuration of the apparatus can be simplified.

(Modification 2 of Embodiment 3)
Next, a nucleic acid extraction apparatus according to Modification 2 of Embodiment 3 will be described with reference to FIG. FIG. 19 is a cross-sectional view of the nucleic acid extraction unit 5 used in the nucleic acid extraction apparatus according to the second modification of the third embodiment. The configuration other than the nucleic acid extraction unit 5 is the same as that in the third embodiment.

  As shown in FIG. 19, the nucleic acid extraction unit 5 in the present modification is similar to the nucleic acid extraction unit 3 in the third embodiment, in the container 10, the first injection port 21, the second injection port 22, and the capturing unit. 30, a retention part 40, a first discharge port 61, a second discharge port 62, and an opening / closing mechanism 80. The opening / closing mechanism 80 includes an elastic member 81 and a movable member 82. The movable member 82 has a heater 70.

  In the nucleic acid extraction unit 5 in this modification, a nucleic acid test reagent for testing the nucleic acid 230 contained in the nucleic acid solution 320 is mixed with the concentrated nucleic acid solution 320 through the capturing unit 30.

  Specifically, as shown in FIG. 19, the nucleic acid test reagent 710 is injected from the nucleic acid test reagent container 700 into the container 10. The nucleic acid test reagent 710 introduced from the nucleic acid extraction reagent container 300 is injected into the first container part 11 through the second injection port 22. Specifically, a nucleic acid test is conducted from the nucleic acid test reagent container 700 into the first container portion 11 by passing the injection needle of the nucleic acid test reagent container 700 through a rubber plug 22a provided in the second injection port 22 (through hole). Reagent 710 is injected. The nucleic acid test reagent 710 injected into the first container unit 11 passes through the capture unit 30 and is mixed with the nucleic acid solution 320 retained in the retention unit 40. That is, in this modification, the nucleic acid test reagent 710 is mixed with the concentrated nucleic acid solution 320 in the container 10.

  Further, when the nucleic acid test reagent 710 is injected into the container 10, the inside of the container 10 may be decompressed (slightly decompressed) by exhausting the air in the container 10 from the first outlet 61 by the vacuum pump 101.

  As described above, according to the nucleic acid extraction apparatus of the present modification, the nucleic acid extraction reagent 5 is mixed with the nucleic acid test reagent 710 in the concentrated nucleic acid solution 320. Thereby, since the volume of the concentrated nucleic acid solution 320 can be increased, when the concentrated nucleic acid solution 320 is fed, it adheres to the wall surface of the container 10 or the piping, and the amount of the nucleic acid solution 320 is lost. Can be suppressed.

  Furthermore, according to this modification, even if the nucleic acid 230 remains in the capture unit 30, the nucleic acid 230 remaining in the capture unit 30 can be passed through the capture unit 30 by injecting the nucleic acid test reagent 710. . Thereby, the nucleic acid 230 remaining in the capturing unit 30 can be mixed with the concentrated nucleic acid solution 320 together with the nucleic acid test reagent 710. Accordingly, the concentration of the nucleic acid solution 320 can be further increased.

  Moreover, in this modification, as shown in FIG. 20, the nucleic acid test reagent 710 and the nucleic acid solution 320 injected into the container 10 may be agitated by vibrating the elastic member 81 by the movable member 82. For example, after injecting the nucleic acid test reagent 710 into the container 10, the nucleic acid solution 320 can be mechanically vibrated and agitated by vibrating the elastic member 81 by moving the movable member 82 up and down in small increments. Thereby, the nucleic acid test reagent 710 and the nucleic acid solution 320 can be easily stirred and mixed with a simple configuration without using a special stirring member. Note that the elastic member 81 may be vibrated by ultrasonic vibration or the like instead of the vertical movement of the movable member 82.

  Further, when the nucleic acid solution 320 is stirred, a vibrating rod 83 for vibrating the elastic member 81 may be incorporated in the movable member 82B of the opening / closing mechanism 80B as in the nucleic acid extraction unit 6 shown in FIG. Good. In this case, even when the elastic member 81 is vibrated by the vibrating rod 83, the elastic member 81 is pressed by the cylindrical portion of the movable member 82B. Thereby, when the elastic member 81 is vibrated, the nucleic acid solution 320 can be prevented from leaking from the gap between the second discharge port 62 and the elastic member 81.

  Note that this modification can be applied not only to the third embodiment and its modifications, but also to the first and second embodiments and these modifications.

  In the present modification, the nucleic acid test reagent 710 is injected into the container 10 from the second injection port 22, but is not limited to this, and from another injection port other than the second injection port 22 into the container 10. It may be injected. Such another injection port may be provided on the upper wall of the container 10 or may be provided on the side wall of the container 10.

(Modification 3 of Embodiment 3)
Next, a nucleic acid extraction apparatus according to Modification 3 of Embodiment 3 will be described with reference to FIG. FIG. 22 is a cross-sectional view of the nucleic acid extraction unit 7 used in the nucleic acid extraction apparatus according to the third modification of the third embodiment. The configuration other than the nucleic acid extraction unit 7 is the same as that in the third embodiment.

  As shown in FIG. 22, the nucleic acid extraction unit 7 in the present modification is similar to the nucleic acid extraction unit 3 in the third embodiment in that the container 10, the first injection port 21, the second injection port 22, and the capturing unit 30, a retention part 40, a first discharge port 61, a second discharge port 62, and an opening / closing mechanism 80. The opening / closing mechanism 80 includes an elastic member 81 and a movable member 82. The movable member 82 has a heater 70.

  The nucleic acid extraction unit 7 in the present modification further includes a liquid amount sensor 90 for detecting the liquid amount of the liquid staying in the staying portion 40. The liquid amount sensor 90 is an optical sensor, for example. Moreover, the liquid quantity sensor 90 can measure the liquid quantity of the nucleic acid solution 320 staying in the staying part 40, for example.

  Thus, the nucleic acid extraction unit 7 in this modification includes the liquid amount sensor 90. Thereby, when the nucleic acid solution 320 is heated, the concentration amount of the nucleic acid solution 320 can be sensed, so that it can be controlled not to be insufficiently concentrated or excessively concentrated. Therefore, the concentration of the nucleic acid solution 320 by heating can be performed efficiently and accurately.

  Note that this modification can be applied not only to the third embodiment and its modifications, but also to the first and second embodiments and these modifications.

(Modification 4 of Embodiment 3)
Next, a nucleic acid extraction apparatus according to Modification 4 of Embodiment 3 will be described with reference to FIGS. 23 and 24 are cross-sectional views of the nucleic acid extraction unit 8 used in the nucleic acid extraction apparatus according to the fourth modification of the fourth embodiment. FIG. 23 shows a state when the elastic member 81 is elastically deformed, and FIG. 24 shows a state when the elastic member 81 is not elastically deformed. The configuration other than the nucleic acid extraction unit 8 is the same as that in the third embodiment.

  As shown in FIGS. 23 and 24, the nucleic acid extraction unit 8 in the present modification is similar to the nucleic acid extraction unit 3 in the third embodiment in that the container 10, the first injection port 21, the second injection port 22, The capturing unit 30, the staying unit 40, the first discharge port 61, the second discharge port 62, and the opening / closing mechanism 80 are provided.

  The nucleic acid extraction unit 8 in this modification is integrated with a test chip 107 in which a micro flow path 107a that is a test flow path is formed. The test chip 107 is a PCR chip for performing nucleic acid amplification by flow PCR, for example.

  Further, in the present modification, the container 10 is provided with a sample insertion cup 200, a nucleic acid extraction reagent container 300, and a nucleic acid test reagent container 700. Further, the container 10 is provided with a spoid 800 for stirring.

  As shown in FIG. 23, in the present modification, as in the third embodiment, the movable member 82 is moved upward to apply pressure to the elastic member 81 to elastically deform the elastic member 81. The discharge port 62 is closed by the elastic member 81. Further, in this state, the nucleic acid solution 320 can be concentrated by heating the nucleic acid solution 320 retained in the retention unit 40.

  On the other hand, as shown in FIG. 24, the second discharge port 62 is opened by moving the movable member 82 downward and returning the elastic member 81 to the original planar state, as in the third embodiment. As a result, the nucleic acid solution 320 in the retention portion 40 is discharged from the container 10 through the through hole 81 a provided in the elastic member 81, and is sent to the microchannel 107 a of the test chip 107.

  The nucleic acid solution 320 sent to the test chip 107 is mixed with a nucleic acid test reagent in the microchannel 107a to become a test solution, and the nucleic acid 230 contained in the test solution is amplified by flow PCR.

  As described above, according to the nucleic acid extraction apparatus in this modification, the same effects as those of the third embodiment can be obtained. That is, since the movable member 82 for elastically deforming the elastic member 81 for opening and closing the second discharge port 62 includes the heater 70, the apparatus configuration can be simplified. Moreover, since the movable member 82 (heater 70) can be brought close to the nucleic acid solution 320 via the film-like elastic member 81, the nucleic acid solution 320 can be efficiently heated.

  Further, in this modification, the container 10 and the test chip 107 are integrated, so that the process for capturing the measurement object 220 from the sample stock solution 210 and the nucleic acid extraction process for extracting the nucleic acid 230 from the captured measurement object 220 are performed. Can be carried out without the work of transferring to a container for each treatment. At this time, since the second discharge port 62 is opened and closed by the elastic member 81, the pretreatment in the container 10 and the feeding of the nucleic acid solution 320 to the test chip 107 can be performed continuously. Therefore, contamination of the test solution by impurities such as bacteria different from the object to be measured (contamination) can be suppressed, and the working time can be shortened. Moreover, since it is not necessary to measure the nucleic acid solution 320, the apparatus configuration can be simplified.

  Further, it is preferable that the volume of the region where the nucleic acid extraction reagent 310 and the object 220 to be measured are subjected to the nucleic acid extraction reaction is as small as possible. For this reason, in this modification, the volume of the 1st container part 11 is small. On the other hand, when the volume of the first container portion 11 is small, it is difficult to stir the nucleic acid extraction reagent 310 and the measurement object 220 by vibration. Therefore, in this modification, the nucleic acid extraction reagent 310 can be easily stirred by sending air to the nucleic acid extraction reagent 310 held on the capturing unit 30 by the dropoid 800. Thereby, the nucleic acid 230 can be easily extracted from the measurement object 220.

(Other variations)
The nucleic acid extraction apparatus 100 and the nucleic acid extraction method according to the present invention have been described based on the first to third embodiments, but the present invention is not limited to the first to third embodiments.

  For example, in the first to third embodiments, the sample stock solution 210 is directly passed through the capturing unit 30, but the present invention is not limited thereto. For example, the sample stock solution 210 may be passed through a prefilter as a pretreatment before the sample stock solution 210 is passed through the capturing unit 30. As an example, a pipe having a prefilter may be inserted between the specimen insertion cup 200 and the first inlet 21. In this way, by passing the specimen stock solution 210 through the prefilter before passing through the capturing unit 30, unnecessary components having a large size can be removed. Thereby, the measurement object 220 can be efficiently captured from the sample stock solution 210.

  In addition, a form obtained by making various modifications conceived by those skilled in the art with respect to each embodiment and modification, and any combination of components and functions in the embodiment without departing from the spirit of the present invention Forms to be made are also included in the present invention.

1, 1A, 1B, 1C, 1D, 1E, 1F, 2, 3, 4, 5, 6, 7, 8 Nucleic acid extraction unit 10 Container 21 First inlet 22 Second inlet 30 Capture section 31 Filter section 32 Support Part 40 Retention part 50 Heating part 61 First outlet 62 Second outlet 80 Opening / closing mechanism 81 Elastic member 82 Movable member 100 Nucleic acid extraction device 106 Pump part 106a Bubble 210 Sample stock solution 220 Measured object 230 Nucleic acid 310 Nucleic acid extraction reagent 320 Nucleic acid Solution 710 Nucleic acid test reagent

Claims (12)

A container,
A first inlet for injecting a sample stock solution into the container;
A capture unit for capturing at least a part of the sample contained in the sample stock solution injected from the first injection port by filtration, at least part of which is disposed in the container;
A second injection port for injecting a nucleic acid extraction reagent for extracting nucleic acid from the object to be measured captured by the capturing unit into the container;
A retention part provided in the container, in which a nucleic acid solution containing a nucleic acid extracted from the object to be measured is retained;
A heating unit for heating the nucleic acid solution retained in the retention unit;
A first outlet for discharging the vapor of the solvent generated when the nucleic acid solution is heated by the heating unit from the container;
The first discharge port also serves as an exhaust port for exhausting air in the container when the inside of the container is decompressed.
Nucleic acid extraction device. And a second outlet for discharging the nucleic acid solution from the container.
The nucleic acid extraction apparatus according to claim 1. An opening / closing mechanism for opening or closing the second outlet;
The opening / closing mechanism has a film-like elastic member that opens or closes the second discharge port by elastic deformation, and a movable member for elastically deforming the elastic member,
The movable member includes the heating unit.
The nucleic acid extraction apparatus according to claim 2. An opening / closing mechanism for opening or closing the second outlet;
The opening / closing mechanism has a film-like elastic member that opens or closes the second discharge port by elastic deformation, and a movable member for elastically deforming the elastic member,
The elastic member is the heating unit.
The nucleic acid extraction apparatus according to claim 2. The nucleic acid test reagent and the nucleic acid solution injected into the container are agitated by vibrating the elastic member by the movable member.
The nucleic acid extraction apparatus according to claim 3 or 4. The heating unit includes an induction heater having a magnetic body and a coil that winds the magnetic body.
The nucleic acid extraction apparatus according to any one of claims 1 to 5. The capturing unit includes a filter unit for filtering and capturing the object to be measured, and a metal support unit that supports the filter unit,
The heating unit is thermally connected to the support unit.
The nucleic acid extraction apparatus according to any one of claims 1 to 6. Having a pump part for introducing bubbles into the nucleic acid solution retained in the retention part,
The nucleic acid extraction apparatus according to any one of claims 1 to 7. The bubbles are hot air,
The nucleic acid extraction apparatus according to claim 8. Furthermore, a liquid amount sensor for detecting the liquid amount of the liquid staying in the staying portion is provided.
The nucleic acid extraction apparatus according to any one of claims 1 to 9. The object to be measured is captured by filtration from the sample stock solution by aspirating the sample stock solution injected into the container by depressurizing the inside of the container by exhausting the air in the container from a discharge port provided in the container. Captured in the department,
Injecting a nucleic acid extraction reagent into the container and extracting the nucleic acid from the measurement object captured by the capturing unit,
The nucleic acid solution containing the extracted nucleic acid is retained in the retention part through the capture part,
Heating the nucleic acid solution staying in the staying part under reduced pressure, and concentrating the nucleic acid solution by discharging the vapor of the solvent of the nucleic acid solution from the outlet to the outside of the container;
Nucleic acid extraction method. A nucleic acid test reagent is mixed with the concentrated nucleic acid solution through the capture unit.
The nucleic acid extraction method according to claim 11.

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