WO2017134906A1

WO2017134906A1 - Sample detection device and sample detection method

Info

Publication number: WO2017134906A1
Application number: PCT/JP2016/084910
Authority: WO
Inventors: 雅之小野; 糸長　誠; 祐一長谷川; 辻田　公二; 茂彦岩間
Original assignee: 株式会社Ｊｖｃケンウッド
Priority date: 2016-02-01
Filing date: 2016-11-25
Publication date: 2017-08-10
Also published as: JP2017138112A

Abstract

A sample detection device 50 is provided with a unit retention part 60, a magnetic field generating part 70, and a drive part 80. The unit retention part 60 retains a specimen analysis container 10 for capturing magnetic fine particles bonded to a detection object substance and storing a solution (extraction liquid 105) for extracting an included substance from the detection object substance. The magnetic field generating part 70 has a magnet 72 for capturing the magnetic fine particles in the specimen analysis container 10. The drive part 80 drives the magnetic field generating part 70 so as to cause the magnet 72 to approach the specimen analysis container 10 so that magnetic fine particles released from bonding with the detection object substance by the solution stored in the specimen analysis container 10 are captured in the specimen analysis container 10 by the magnetic force of the magnet 72.

Description

Specimen detection apparatus and specimen detection method

The present disclosure relates to a sample detection apparatus and a sample detection method for detecting a membrane vesicle and its inclusion substance from a sample.

It is widely performed to quantitatively analyze the discovery of a disease or the effect of treatment by detecting and analyzing a specific antigen (or antibody) associated with the disease from a specimen as a biomarker. In recent years, minute membrane vesicles called exosomes have attracted attention. Membrane vesicles such as exosomes have a structure in which an encapsulated substance is wrapped with a membrane. The exosome is sometimes called a microvesicle or an extracellular vesicle, and there are a plurality of names.

Exosomes are contained in body fluids such as blood, lymph, saliva, urine, breast milk and semen. Exosomes are roughly spherical in liquid and are covered with a lipid bilayer. In general, exosomes contain multiple proteins of different types. Among them, the protein to be recognized is a protein expressed on the surface of the lipid bilayer membrane, such as a transmembrane protein, an adhesion molecule, a membrane transport protein, a membrane fusion protein, a glycoprotein, and the like.

Exosomes are very small, about 100 nm in diameter, so it is difficult to optically detect exosomes directly.

In Patent Document 1, exosomes are sandwich-captured from a specimen using a sample analysis disk and a label such as fine particles using an antibody that specifically reacts with a protein (antigen) expressed on the surface of the exosome. A specimen detection method is described in which exosomes are indirectly detected by optical detection.

JP 2013-134083 A

On the other hand, macromolecular biological substances such as DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and miRNA (microRNA), which are substances that transmit cellular information, become encapsulated in exosomes and metastasize in the body. Some studies have been reported. Thus, attention has been focused on the correlation between the number of exosomes in body fluids and the number of encapsulated substances and a serious disease such as cancer or Alzheimer's. In general, since the reaction of a biological substance may be low in stability, it is desirable to perform exosome detection (measurement) and inclusion substance detection (measurement) on the same specimen.

It is possible to count exosomes with high accuracy by the method described in Patent Document 1. In order to count exosomes, it is necessary to dry the specimen. Therefore, in order to extract the exosome-encapsulating substance, it is necessary to react the exosome captured on the substrate surface with the reagent again after counting the exosome.

Also, exosomes and inclusion substances may be denatured by drying. Therefore, it is necessary to take a method of counting exosomes and extracting encapsulated substances separately and evaluating the relationship with each result. Therefore, conventionally, exosome detection and inclusion substance detection are performed using separate specimens. Furthermore, minimally invasive diagnosis is desired in which examination or measurement is performed with as little sample as possible.

An object of the embodiment is to provide a specimen detection apparatus and a specimen detection method capable of performing detection of membrane vesicles such as exosomes and detection of encapsulated substances using the same specimen.

According to the first aspect of the embodiment, the unit holding unit that holds the sample analysis container that captures the magnetic fine particles that bind to the detection target substance and stores a solution for extracting the inclusion substance from the detection target substance; A magnetic field generator having a magnet for capturing the magnetic fine particles in the sample analysis container, and magnetic fine particles released from binding to the detection target substance by the solution stored in the sample analysis container. A specimen detection apparatus comprising: a drive unit that drives the magnetic field generation unit so that the magnet is brought close to the sample analysis container so as to be captured in the sample analysis container by the magnetic force of the magnet. Provided.

According to the second aspect of the embodiment, the magnetic fine particles that bind to the detection target substance are captured in the sample analysis container, and the magnetic field is maintained so that the magnetic fine particles are captured in the sample analysis container. The sample detection method is characterized in that the inclusion substance is extracted from the detection target substance in a state where the magnetic fine particles are captured in the sample analysis container by the magnetic field.

According to the sample detection apparatus and the sample detection method of the embodiment, detection of membrane vesicles such as exosomes and detection of encapsulated substances thereof can be performed with the same sample.

FIG. 1 is a top view showing a sample analysis container according to an embodiment. FIG. 2 is a cross-sectional view showing the AA cross section of the sample analysis container of FIG. FIG. 3 is an enlarged perspective view showing a state where the well of FIG. 1 is cut along BB. FIG. 4 is a top view showing the sample detection apparatus of one embodiment. FIG. 5 is a cross-sectional view showing a CC cross section and a DD cross section of the specimen detection apparatus of FIG. FIG. 6 is a top view showing the holding plate of the unit holding portion. FIG. 7 is a top view showing the base substrate of the unit holding part. FIG. 8 is a top view and a cross-sectional view taken along the line AA showing the magnetic field generation unit. FIG. 9 is a flowchart for explaining the sample detection method according to the embodiment. FIG. 10 is a cross-sectional view showing a state in which a buffer solution containing an antibody is injected into a well of a sample analysis container. FIG. 11 is a schematic cross-sectional view showing a state in which the antibody is fixed to the track region of the sample analysis disk. FIG. 12 is a schematic cross-sectional view showing a state in which the block layer is formed in the track region. FIG. 13 is a cross-sectional view showing a state in which the sample liquid of the specimen to be detected is injected into the well of the sample analysis container. FIG. 14 is a schematic cross-sectional view showing an exosome. FIG. 15 is a schematic cross-sectional view showing a state in which the exosome is captured in the recess in the track region. FIG. 16 is a cross-sectional view showing a state in which a buffer solution containing magnetic fine particles is injected into the well of the sample analysis container. FIG. 17 is a schematic cross-sectional view showing a state in which exosomes captured in the recesses in the track region and magnetic fine particles are combined. FIG. 18 is a cross-sectional view showing a state where an extract for releasing the inclusion substance from the exosome is injected into the well with the magnet approaching the bottom surface of the well. FIG. 19 is a schematic cross-sectional view showing a state in which magnetic fine particles are captured in the recesses in the track region by the magnetic force of the magnet. FIG. 20 is a top view showing a sample analysis disk in which a plurality of reaction regions are formed.

[Sample analysis container]
A sample analysis container according to an embodiment will be described with reference to FIGS.

FIG. 1 shows the sample analysis container as viewed from the cartridge side. FIG. 2A shows an AA cross section of the sample analysis container of FIG. FIG. 2 (b) shows that the cartridge can be removed from the sample analysis disk. FIG. 3 shows a partially enlarged view of the well of FIG. 1 cut along BB.

1 and 2A, the sample analysis container 10 includes a sample analysis disk 20 and a cartridge 30. As shown in FIG. The sample analysis container 10 is one form of a sample analysis container for capturing membrane vesicles such as exosomes, which are detection target substances, and magnetic fine particles that label the membrane vesicles.

The sample analysis disc 20 has a disk shape equivalent to an optical disc such as a Blu-ray disc (BD), DVD, or compact disc (CD). The sample analysis disk 20 is made of, for example, a resin material such as polycarbonate resin or cycloolefin polymer generally used for optical disks.

As shown in FIG. 3, on the surface of the sample analysis disk 20, track regions 23 in which convex portions 21 and concave portions 22 are alternately arranged in the radial direction are formed. The convex portion 21 and the concave portion 22 are formed in a spiral shape from the inner peripheral portion toward the outer peripheral portion. The convex portion 21 corresponds to a land of the optical disc. The recess 22 corresponds to a groove of the optical disc.

As shown in FIG. 1, the cartridge 30 has a ring shape. A plurality of cylindrical through holes 31 are formed in the cartridge 30 in the circumferential direction. The plurality of through holes 31 are formed at equal intervals so that the centers thereof are located on the same circumference.

2A and 3, the sample analysis container 10 includes a plurality of wells 40 formed by the through holes 31 of the cartridge 30 and the track regions 23 of the sample analysis disk 20. The plurality of wells 40 are formed at equal intervals so that their centers are located on the same circumference. The inner peripheral surface of the through hole 31 constitutes the inner peripheral surface of the well 40, and the track region 23 of the sample analysis disk 20 constitutes the bottom surface of the well 40. The well 40 is a container for storing a solution such as a sample solution or a buffer solution.

By placing a packing made of an elastically deformable member such as silicone rubber between the through hole 31 and the sample analysis disk 20, the possibility of solution leakage can be reduced.

As shown in FIG. 2B, the cartridge 30 and the sample analysis disk 20 can be separated. Detection of membrane vesicles such as exosomes, which are detection target substances, specifically, detection of magnetic fine particles labeling the membrane vesicles is performed by the sample analysis disk 20 alone from which the cartridge 30 is separated.

[Specimen detection device]
A sample detection apparatus according to an embodiment will be described with reference to FIGS.

FIG. 4 shows a sample analysis container, a unit holder, and a magnetic field generator. FIG. 5A shows a cross section taken along the line CC of FIG. FIG. 5A shows a state in which the magnetic field generator is close to the sample analysis disk. FIG. 5B shows a state in which the magnetic field generator is separated from the sample analysis disk and the unit holder. FIG. 5C shows a cross section taken along line DD of FIG. FIG. 6 shows a holding plate of the unit holding part. FIG. 7 shows the base substrate of the unit holding part. FIG. 8A shows a magnetic field generator. FIG. 8B shows a cross section taken along line AA of FIG. 6, FIG. 7, and FIG. 8 (a) correspond to FIG. FIG. 8B corresponds to FIG.

4 and 5A, the specimen detection apparatus 50 includes a unit holding unit 60, a magnetic field generation unit 70, and a driving unit 80.

The unit holding unit 60 includes a pressing plate 61 and a base substrate 62. The unit holding unit 60 holds the sample analysis container 10 by holding the sample analysis container 10 between the holding plate 61 and the base substrate 62.

As shown in FIGS. 4 and 5C, the pressing plate 61 and the base substrate 62 may be fixed by screws 65 or the like via spacers 64. It is desirable that the height of the spacer 64 be equal to or slightly lower than the height of the sample analysis container 10.

As shown in FIG. 6, the pressing plate 61 has an opening 61a. As shown in FIG. 4, in a state where the sample analysis container 10 is held by the unit holder 60, the openings 61a have an opening diameter such that all the wells 40 are located in the openings 61a.

As shown in FIG. 7, the base substrate 62 has a plurality of through holes 62a. As shown in FIG. 5B, the through hole 62 a is formed corresponding to the well 40. Specifically, the through hole 62 a is formed to be positioned on the bottom surface of the well 40 in a state where the sample analysis container 10 is held by the unit holding unit 60. That is, the plurality of through holes 62a are formed at equal intervals so that their centers are located on the same circumference.

8A and 8B, the magnetic field generation unit 70 includes a base 71 and a plurality of magnets 72. The magnet 72 is fixed to the base 71.

4 and 5A, the magnet 72 is formed corresponding to the well 40. As shown in FIG. Specifically, the magnet 72 is formed so as to be positioned on the bottom surface of the well 40 in a state where the sample analysis container 10 is held by the unit holding unit 60. That is, the plurality of through holes 62a are formed at equal intervals so that their centers are located on the same circumference. The through hole 62 a of the base substrate 62 is an insertion hole for allowing the magnet 72 to approach the bottom surface of the well 40. Therefore, the through hole 62a has a hole diameter slightly larger than the outer diameter of the magnet.

As shown in FIGS. 5A and 5B, the driving unit 80 moves the magnetic field generating unit 70 so that the magnet 72 approaches or separates from the bottom surface of the well 40 of the sample analysis container 10. Drive. A motor may be used as the drive unit 80. By bringing the magnet 72 close to the bottom surface of the well 40, a magnetic field can be generated in the well 40 of the sample analysis container 10.

[Sample detection method]
A sample detection method according to an embodiment will be described with reference to the flowchart shown in FIG. 9 and FIGS. 10 to 20. In the sample detection method of the present embodiment, an exosome will be described as an example of a membrane vesicle that is a detection target substance.

In step S1, the operator attaches the sample analysis container 10 to the unit holding unit 60 of the sample detection device 50 as shown in FIG. The operator injects the buffer 100 containing the antibody 101 (first binding substance) into the well 40 of the sample analysis container 10.

The operator incubates the sample analysis container 10 at an appropriate temperature for an appropriate time. For example, it is allowed to incubate at 4 ° C. overnight, which is used for general immunoassay. As a result, as shown in FIG. 11, the antibody 101 is fixed on the track area 23 of the sample analysis disk 20.

The operator discharges the buffer solution 100 from the well 40 and cleans the well 40 with the buffer solution. The antibody 101 not fixed to the track region 23 is removed by this washing.

Although FIG. 10 shows a state in which the magnet 72 of the magnetic field generating unit 70 is close to the bottom surface of the well 40, in step S1, the magnet 72 may be separated from the bottom surface of the well 40.

Step S1 is a process required when the operator fixes the antibody 101. When using the sample analysis container 10 or the sample analysis disk 20 to which the antibody 101 is fixed in advance in a factory or the like, step S1 can be omitted.

As shown in FIG. 11, the antibody 101 is fixed to the convex portion 21 and the concave portion 22 constituting the track region 23 by hydrophobic bonding. The method for immobilizing the antibody 101 is not limited to hydrophobic binding. An appropriate chemical treatment may be performed on the track region 23 to fix the antibody 101 to the track region 23 using a covalent bond or the like. As a method for fixing the antibody 101 to the track region 23, a method generally used in an immunological assay can be used.

In step S2, the operator performs a blocking process on the inside of the well 40 in order to prevent nonspecific adsorption of the antigen other than the antigen identification part of the antibody 101. Specifically, the operator injects skim milk diluted with a buffer into the well 40 and shakes the sample analysis container 10 for an appropriate time.

Skimmed milk contains a protein that does not adhere to exosomes and is suitable for blocking treatment. In addition, the substance used for the blocking treatment is not limited to skim milk as long as it has the same effect.

The operator discharges the buffer solution containing skim milk from the well 40 and cleans the well 40 with the buffer solution. As a buffer used for washing, skim milk may or may not be contained. It is also possible to omit the cleaning.

Thereby, the block layer 102 is formed on the track region 23 as shown in FIG.

In step S3, the operator injects the sample solution 103 (first solution), which is the specimen to be detected, into the well 40 as shown in FIG. Note that the sample solution 103 may not include the exosome to be detected. Hereinafter, the case where the sample solution 103 contains exosomes to be detected will be described.

The sample solution 103 contains exosomes 90 to be detected. As shown in FIG. 14, the exosome 90 is covered with a lipid bilayer membrane 91. In the lipid bilayer membrane 91, a plurality of types of proteins such as transmembrane proteins exist as surface molecules 92. The number of proteins or the position in the lipid bilayer membrane 11 varies depending on the type of exosome and also varies depending on the individual. These surface molecules 92 are used as antigens to recognize exosomes 90 using an antigen-antibody reaction. The existence of various proteins such as CD63, CD9, and Rab-5b as surface molecules 92 that are antigens has been reported in many papers.

In the exosome 90, an encapsulated substance 93, which is a macromolecular biomaterial such as DNA, RNA, miRNA and the like, which is a substance that transmits cell information, is encapsulated in a lipid bilayer membrane 91. The diameter of the exosome 90 is about 100 nm.

The operator incubates the sample analysis container 10 at an appropriate temperature for an appropriate time. You may shake at the time of incubation. For example, the sample analysis container 10 is shaken at 37 ° C. for about 2 hours.

Thereby, the surface molecule 92 of the exosome 90 and the antibody 101 immobilized on the track region 23 are specifically bound by the antigen-antibody reaction. As a result, as shown in FIG. 15, the exosome 90 is captured in the track region 23, specifically, the recess 22 of the track region 23.

The operator discharges the sample solution 103 from the well 40 and cleans the well 40 with a buffer solution. The exosomes that are not bound to the antibody 101 and are dispersed in the sample solution 103 and the exosomes that are attached to the track region 23 by non-specific adsorption that is not an antigen-antibody reaction are removed by this washing. That is, the exosome 90 to be detected is captured in the concave portion 22 of the track region 23, and the exosome not to be detected is removed by washing.

Although FIG. 13 shows a state in which the magnet 72 of the magnetic field generation unit 70 is close to the bottom surface of the well 40, the magnet 72 may be separated from the bottom surface of the well 40 in steps S2 and S3.

In step S4, the operator injects into the well 40 a buffer solution 104 (second solution) containing magnetic fine particles (magnetic beads) 110 to be labeled as shown in FIG. The operator causes the sample analysis container 10 to incubate at an appropriate temperature for an appropriate time.

As shown in FIG. 17, the magnetic fine particles 110 are formed of a synthetic resin such as polystyrene formed in a substantially spherical shape. The magnetic fine particles 110 contain a magnetic material 111 such as iron oxide. An antibody 112 (second binding substance) that specifically binds to the surface molecule 92 of the exosome 90 is immobilized on the surface of the magnetic fine particle 110. The diameter of the magnetic fine particle 110 is about 200 nm.

The magnetic fine particles 110 are magnetized by the magnetic field of the magnet 72 and move toward the bottom surface of the well 40. The surface molecule 92 of the exosome 90 and the antibody 112 of the magnetic particle 110 are specifically bound by an antigen-antibody reaction. As a result, the magnetic fine particles 110 are captured in the concave portion 22 of the track region 23 in the sample analysis container 10, specifically, the well 40, in a state of being coupled to the exosome 90.

16 shows a state in which the magnet 72 of the magnetic field generating unit 70 is close to the bottom surface of the well 40, but in a step S4, the magnet 72 may be separated from the bottom surface of the well 40. In step S <b> 4, the magnetic particles 110 can be quickly moved toward the bottom surface of the well 40 by bringing the magnet 72 close to the bottom surface of the well 40. Thereby, step S4 can be shortened.

The operator operates the specimen detection device 50 so that the magnet 72 is separated from the bottom surface of the well 40 when the magnet 72 is close to the bottom surface of the well 40. The operator drains the buffer solution 104 from the well 40 and cleans the well 40 with the buffer solution. The magnetic fine particles 110 that are not bonded to the exosome 90 and are dispersed in the buffer 104 are removed by this washing. That is, the magnetic fine particles 110 that are bonded to the detection target exosome 90 are captured in the recesses 22 of the track region 23, and excess magnetic fine particles 110 that are not bonded to the detection target exosome 90 are removed by washing.

In step S5, the operator operates the specimen detection apparatus 50 so that the magnet 72 approaches the bottom surface of the well 40 as shown in FIG. The operator injects the extraction liquid 105 (third solution) for releasing the inclusion substance 93 from the exosome 90 into the well 40.

As shown in FIG. 19, the inclusion substance 93 can be liberated in the extract 105 by dissolving the exosome 90 in the extract 105. The time for releasing the inclusion substance 93 from the exosome 90 is, for example, about several minutes after the extraction liquid 105 is injected.

When the exosome 90 is dissolved, the bond between the exosome 90 and the magnetic fine particle 110 captured in the recess 22 of the track region 23 is released. Since the magnetic fine particles 110 having been released from the coupling are attracted toward the magnet 72 by the magnetic force of the magnet 72, the magnetic particles 110 are captured in the sample analysis container 10, specifically, in the recess 22 of the track region 23 in the well 40. maintain. That is, the drive unit 80 causes the magnet 72 to approach the bottom surface of the well 40, thereby capturing the magnetic fine particles 110 in the recess 22 on the bottom surface in the well 40 by the magnetic force of the magnet 72.

In step S6, the operator extracts the extracted solution 105 from which the inclusion substance 93 has been released from the well 40 using a pipette or the like, and analyzes the inclusion substance 93 using, for example, a real-time PCR (Real-Time Polymerase Chain Reaction) method. To do. The real-time PCR method is a method in which DNA is amplified using an enzyme reaction by a DNA polymerase, and an increase amount of an amplified product is monitored and analyzed in real time.

For example, an amplification curve of an amplification product is obtained by applying a real-time PCR method to a reference sample diluted stepwise. A calibration curve is created by calculating a Ct (Threshold Cycle) value, which is a point where the amplification curve intersects with the set threshold value. The Ct value of the extract 105 extracted from the well 40 can be calculated similarly to the reference sample, and the DNA that is the inclusion substance 93 of the exosome 90 can be qualitatively and quantified from the calculated Ct value and the created calibration curve.

Therefore, the inclusion substance 93 in each well 40 can be analyzed by sampling the extract 105 for each well 40.

The operator discharges the remaining extract 105 from the well 40 with the magnet 72 approaching the bottom surface of the well 40, and cleans the well 40 with a buffer solution. The operator dries the well 40. The drying time is about 10 to 15 minutes. Drying may be such that water drops on the bottom surface of the well 40 disappear.

Since the magnet 72 is close to the bottom surface of the well 40 during the period in which the well 40 is washed and dried, the magnetic fine particle 110 maintains the state of being captured in the recess 22 by the magnetic force of the magnet 72.

The operator removes the sample analysis container 10 from the unit holder 60 of the specimen detection apparatus 50 in step S7. The operator removes the cartridge 30 of the sample analysis container 10 from the sample analysis disk 20 as shown in FIG. The dried magnetic fine particles 110 maintain a state where they are captured in the recesses 22 by van der Waals force.

2B and 20, a plurality of reaction regions 24 are formed in the sample analysis disk 20 corresponding to the plurality of wells 40. That is, the plurality of reaction regions 24 are formed at equal intervals so that their centers are located on the same circumference. Magnetic fine particles 110 are trapped on the recesses 22 of the reaction region 24 (see FIG. 19).

The operator optically detects and counts the magnetic fine particles 110 that are the labels captured on the recesses 22 of the reaction region 24 using, for example, an optical pickup or the like, thereby counting the exosomes captured on the recesses 22. 90 can be indirectly detected and counted.

For example, laser light is irradiated from the optical pickup arranged outside to the concave portion 22 of the reaction region 24 where the magnetic fine particles 110 are captured. By analyzing the reflected light from the reaction region 24, the magnetic fine particles 110 can be detected and counted.

Specifically, the optical pickup includes an objective lens for condensing the laser beam on the track area 23. The sample analysis disk 20 is rotated in the same manner as a general optical disk, and the optical pickup is moved in the radial direction of the sample analysis disk 20 so that the laser beam condensed by the objective lens is tracked (specifically, a concave portion). Scan along 22). From the detection signal obtained by the reflected light from the reaction region 24, the magnetic fine particles 110 captured in the recess 22 are detected and counted.

In the sample detection apparatus and the sample detection method of the present embodiment, the exosome 90 is captured in the concave portion 22 of the track region 23 in a state of being coupled to the magnetic fine particle 110. The inclusion substance 93 is released from the exosome 90 by the extract 105 while the magnet 72 is brought close to the bottom surface of the well 40. In a state where the magnetic fine particles 110 are captured in the recess 22 by the magnetic force of the magnet 72, the extracted liquid 105 from which the inclusion substance 93 is released is extracted, and the inclusion substance 93 is analyzed. On the other hand, the sample analysis disk 20 is dried, and the magnetic fine particles 110 trapped in the reaction region 24 are optically detected.

Therefore, according to the sample detection apparatus and the sample detection method of the present embodiment, the detection of the exosome 90 and the detection of the inclusion substance 93 can be performed with the same sample (sample solution 103).

The present invention is not limited to the embodiment described above, and various modifications can be made without departing from the gist of the present invention.

In this embodiment, after injecting the sample solution 103 into the well 40 and capturing the exosome 90 in the recess 22, the buffer solution 104 is injected into the well 40 and the magnetic fine particles 110 are combined with the exosome 90. However, the present invention is not limited to this. Is not to be done. For example, the sample solution 103 and the buffer solution 104 may be injected into the well 40 and mixed, or the sample solution 103 and the buffer solution 104 are mixed in another container such as a microtube or a column, and the mixture solution is added to the well. 40 may be injected.

In this embodiment, the sample analysis container 10 includes the sample analysis disk 20 and the cartridge 30. However, the present invention is not limited to this. A microplate may be used as the sample analysis container 10. By releasing the inclusion substance 93 from the exosome 90 in a state where the magnet 72 is brought close to the bottom surface of the well of the microplate, the same effect as in the present embodiment can be obtained. A test tube such as a macro tube or a petri dish may be used as the sample analysis container 10. When a microplate, a test tube, a petri dish, or the like is used as the sample analysis container 10, the magnetic fine particles 110 only need to be able to maintain the state captured in the sample analysis container 10 by the magnetic force of the magnet 72.

The unit holding unit 60 is not limited to this embodiment as long as the well 40 and the sample analysis disk 20 can be kept in close contact so that the liquid does not leak due to separation. For example, the sample analysis container 10 may be held by various lock mechanisms such as a claw for engaging the cartridge 30 and the sample analysis disk 20. When the sample analysis container 10 is placed on the sample detection device 50, the sample analysis container 10 may be held by a holding structure on the sample analysis container 10 side.

The present invention can be used when detecting a membrane vesicle and its inclusion substance from a specimen.

Claims

A unit holding unit for holding a sample analysis container for capturing a magnetic fine particle that binds to a detection target substance and storing a solution for extracting an inclusion substance from the detection target substance;
A magnetic field generator having a magnet for capturing the magnetic fine particles in the sample analysis container;
The magnet is used for the sample analysis so that the magnetic fine particles released from the binding to the detection target substance by the solution stored in the sample analysis container are captured in the sample analysis container by the magnetic force of the magnet. A drive unit that drives the magnetic field generation unit to approach the container;
A specimen detection apparatus comprising:
2. The specimen detection apparatus according to claim 1, wherein the substance to be detected is a membrane vesicle having a structure in which the inclusion substance is wrapped with a film.
3. The specimen detection apparatus according to claim 2, wherein the membrane vesicle is an exosome.
The sample analysis container comprises:
A cartridge having a through hole;
A sample analysis disk having a track region in which convex portions and concave portions are alternately arranged;
With
The solution is stored in a well constituted by the inner peripheral surface of the through hole and the track region,
The magnet is disposed corresponding to the well;
The specimen detection apparatus according to any one of claims 1 to 3, wherein the magnetic fine particles are captured in a recess of the track region in the well by the magnetic force of the magnet.
Capture the magnetic particles that bind to the detection target substance in the sample analysis container,
Generating a magnetic field so as to maintain the magnetic fine particles captured in the sample analysis container;
An analyte detection method, wherein an inclusion substance is extracted from the detection target substance in a state where the magnetic fine particles are captured in the sample analysis container by the magnetic field.
6. The specimen detection method according to claim 5, wherein the substance to be detected is a membrane vesicle having a structure in which the inclusion substance is wrapped with a film.
The specimen detection method according to claim 6, wherein the membrane vesicle is an exosome.
The sample analysis container comprises:
A cartridge having a through hole;
A sample analysis disk having a track region in which convex portions and concave portions are alternately arranged;
With
The sample analysis disk has a track region in which convex portions and concave portions are alternately arranged,
8. The inclusion substance is extracted from the detection target substance in a state where the magnetic fine particles are trapped in the recess of the track region in the sample analysis container by the magnetic field. 2. The specimen detection method according to claim 1.