JP2020034424A - Sensor cartridge - Google Patents

Abstract

To provide a sensor cartridge with which it is possible to prevent liquid from getting mixed in a passage and improve accuracy of measuring a specific substance such as a biomarker.SOLUTION: A sensor cartridge comprises: a first liquid pool 11 and a third liquid pool 13; a reaction field 20; a first passage 31 connected to the first liquid pool 11 and constituted so as to be capable of transporting liquid from the first liquid pool 11 to above the reaction field 20; and a third passage 33 connected at one end 33-1 to the third liquid pool 13 and connected at the other end 33-2 to the first passage 31 at a second connection point 42. At the second connection point 42, a direction from the one end 33-1 to the other end 33-2 of the third passage 33 has a direction component of the first passage 31 from the first liquid pool 11 to the reaction field 20. A width of the first passage 31 at a downstream portion 42A of the second connection point 42 is larger than a width of the first passage 31 at an upstream portion 42B of the second connection point 42.SELECTED DRAWING: Figure 2

Description

  The present invention relates to a sensor cartridge, and more particularly, to a sensor cartridge for measuring a physical quantity of a specific substance such as a biomarker.

  Conventionally, as shown in FIG. 6, for example, a substantially U-shaped reagent chamber 101 in a plan view that houses a nitrocellulose membrane in which magnetic particles are dispersed, and a substantially U-shaped micro-tube in a plan view connected to the reagent chamber 101. A sensor cartridge 100 including a channel 102, a sample chamber 103 connected to one end of the microchannel 102 and containing a sample to be measured, and a reaction chamber 104 connected to the other end of the microchannel 102 has been proposed. .

  In this sensor cartridge 100, a sample to be measured is supplied from a sample chamber 103 to a reaction chamber 104 via a microchannel 102. When a phosphate buffer solution is supplied from another reagent chamber (not shown) to the reagent chamber 101, the phosphate buffer solution flows through the nitrocellulose membrane in the reagent chamber 101, and phosphoric acid in which the magnetic particles are uniformly dispersed is provided. A salt buffer (hereinafter, also referred to as “magnetic particle dispersion”) is obtained. When the magnetic particle dispersion is supplied from the reagent chamber 101 to the reaction chamber 104 via the microchannel 102, the specific substance contained in the sample to be measured in the reaction chamber 104 biochemically reacts with the magnetic particles, This makes it possible to measure a specific substance with a magnetic sensor (not shown) arranged in the reaction chamber 104.

Chinese Patent Application Publication No. 105628909

  However, in the above-described conventional technique, since the connection point 105 between the reagent chamber 101 and the microchannel 102, which is substantially U-shaped in plan view, has a T shape, the sample to be measured flowing through the microchannel 102 is There is a possibility that a specific substance such as a biomarker contained in the sample to be measured may remain in the reagent chamber 101 after flowing. When a phosphate buffer is supplied to the reagent chamber 101 in a state where the specific substance remains in the reagent chamber 101, a specific substance that biochemically reacts with the magnetic particles appears in the reagent chamber 101 and has already reacted with the specific substance. A magnetic particle dispersion in which mixed magnetic particles and unreacted magnetic molecules are mixed is formed. When such a magnetic particle dispersion liquid is supplied to the reaction chamber 104, the magnetic molecule dispersion liquid containing no magnetic particles reacted with the specific substance is captured on the magnetic sensor as compared with the case where the magnetic particle dispersion liquid is supplied to the reaction chamber 104. In some cases, the number of magnetic particles to be measured changes, and the physical quantity of a specific substance cannot be accurately measured by a magnetic sensor.

  An object of the present invention is to provide a sensor cartridge capable of preventing mixing of liquids in a flow channel and improving measurement accuracy of a specific substance such as a biomarker.

In order to achieve the above object, the present invention provides the following means.
[1] a first liquid pool, a second liquid pool, and a third liquid pool;
A reaction field,
A first flow path connected to the first liquid pool and configured to be able to transport liquid from the first liquid pool onto the reaction field;
A second flow path having one end connected to the second liquid pool and the other end connected to the first flow path at a first connection point;
A third flow path having one end connected to the third liquid pool and the other end connected to the first flow path at a second connection point;
At the second connection point, the direction from the one end of the third flow path to the other end has a directional component from the first liquid pool of the first flow path to the reaction field. And
A sensor cartridge in which a width of the first flow path at a downstream part of the second connection point is larger than a width of the first flow path at an upstream part of the second connection point.
[2] Whether the width of the first flow path at the downstream part of the second connection point is equal to the sum of the width of the first flow path and the width of the third flow path at the upstream part of the second connection point Or the sensor cartridge according to the above [1], which is larger than the total.
[3] At the first connection point, a direction from the one end of the second flow path to the other end is a direction from the first liquid pool of the first flow path to the reaction field. Having components,
The sensor according to [1] or [2], wherein a width of the first flow path at a downstream portion of the first connection point is larger than a width of the first flow path at an upstream portion of the first connection point. cartridge.
[4] Whether the width of the first flow path at the downstream part of the first connection point is the same as the sum of the width of the first flow path and the width of the second flow path at the upstream part of the first connection point Or the sensor cartridge according to the above [3], which is larger than the total.
[5] The second connection point is provided at a position closer to the reaction field than the first connection point in the first flow path,
The above [1] to [1], wherein the first flow path has a curved portion upstream of the second connection point and is connected to the non-curved portion of the third flow path at the second connection point. 4] The sensor cartridge according to any one of [1] to [4].
[6] The above [5], wherein the second flow path has a curved portion on the upstream side of the first connection point, and is connected to the non-curved portion of the first flow path at the first connection point. 4. The sensor cartridge according to 1.
[7] The sensor cartridge according to any of [1] to [6], wherein the first flow path is parallel to the third flow path at the second connection point.
[8] The sensor cartridge according to any one of [1] to [7], wherein the second flow path is parallel to the first flow path at the first connection point.
[9] The sensor cartridge according to any one of [1] to [8], wherein the third liquid pool stores magnetic beads or a detection substrate.

  ADVANTAGE OF THE INVENTION According to this invention, mixing of liquids in a flow path can be prevented, and the measurement precision of specific substances, such as a biomarker, can be improved.

FIG. 2 is a perspective view schematically showing a configuration of a sensor cartridge according to the embodiment of the present invention. FIG. 2 is a plan view illustrating an example of a specific configuration of a channel in the sensor cartridge of FIG. 1. FIG. 2A is a partially enlarged view showing details of a first connection point and a second connection point provided in the flow channel in FIG. 2, and FIG. 2B is a view showing a most preferred embodiment thereof. (A)-(f) is a schematic diagram for demonstrating an example of the usage method of the sensor cartridge of FIG. (A)-(f) is a schematic diagram for demonstrating the modification of the sensor cartridge of FIG. 1, and its usage. FIG. 10 is a plan view illustrating an example of a conventional sensor cartridge.

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.
[Overall configuration of sensor cartridge]
FIG. 1 is a perspective view schematically showing a configuration of the sensor cartridge according to the present embodiment. In the present embodiment, a case where the reaction field is constituted by the surface of the magnetic sensor will be described as an example. In the drawings used in the following description, a characteristic portion may be enlarged for convenience, and the dimensional ratios and the like of the respective components are not limited to those illustrated.

  As shown in FIG. 1, the sensor cartridge 1 of the present embodiment includes a base plate 1A, a substrate 1B laminated on the base plate 1A, and a cover plate 1C mounted on the base plate 1A with the substrate 1B interposed therebetween. It has.

  Specifically, the sensor cartridge 1 is connected to the first liquid pool 11, the second liquid pool 12, the third liquid pool 13, the reaction field 20, and the first liquid pool 11, and The first flow path 31 configured to be able to transport the liquid to the reaction field 20 and one end 32-1 are connected to the second liquid pool 12, and the other end 32-2 is connected to the first connection point. At 41, the second flow path 32 connected to the first flow path 31 and one end 33-1 are connected to the third liquid pool 13, and the other end 33-2 is connected to the second connection point 42 at the second connection point 42. A third flow path 33 connected to the first flow path 31.

  In the present embodiment, the first liquid pool 11, the second liquid pool 12, the third liquid pool 13, and the first flow path 31, the second flow path 32, and the third flow path 33 are provided in the cover plate 1C. A reaction field 20 is provided on a magnetic sensor described below of the substrate 1B. However, the first liquid pool 11, the second liquid pool 12, the third liquid pool 13, and the first flow path 31, the second flow path 32, and the third flow path 33 are not limited to this, and are provided on the base plate 1A. Is also good.

  The first liquid pool 11, the second liquid pool 12, and the third liquid pool 13 are provided on the upper surface 1C-1 of the cover plate 1C, and include a first flow path 31, a second flow path 32, and a third flow path 33. Is provided on the lower surface 1C-2 of the cover plate 1C. The first liquid pool 11, the second liquid pool 12, and the third liquid pool 13 communicate with the first flow path 31, the second flow path 32, and the third flow path 33 through communication holes (not shown), respectively. ing. The sensor cartridge 1 is formed of, for example, a transparent resin, and the first flow path 31, the second flow path 32, and the third flow path 33 are configured to be visible from the upper surface 1C-1 of the cover plate 1C. ing.

  Cap members 51, 52, and 53 formed of elastic members are detachably attached to the first liquid pool 11, the second liquid pool 12, and the third liquid pool 13, respectively. For example, by injecting and storing a predetermined liquid into the first liquid pool 11 using a device such as a syringe with the cap member 51 attached to the first liquid pool 11, and then pressing the cap member 51, The liquid in the one liquid pool 11 is supplied to the first flow path 31. The case where the liquid is stored in the second liquid pool 12 and the third liquid pool 13 is the same as described above, and the description is omitted.

  A reaction field 20 is provided on the upper surface 1B-1 of the substrate 1B, and the reaction field 20 is configured by a surface of a magnetic sensor embedded so that the upper surface is exposed on the substrate 1B. However, the reaction field 20 is not limited to the surface of the magnetic sensor, and may be formed of another member as long as it is a region that can react biochemically with a biomarker (for example, a biomolecule) to be measured.

  Further, the sensor cartridge 1 includes a fourth flow path 34 provided on the downstream side of the reaction field 20 and a drainage pool 14 connected to the fourth flow path 34 through the hole 21 provided in the substrate 1B. Prepare. In the present embodiment, the fourth flow path 34 is provided on the lower surface 1C-2 of the cover plate 1C, and the drainage pool 14 is provided on the upper surface 1A-1 of the base plate 1A. The fourth flow path 34 is not directly connected to the first liquid pool 11, but is directly connected only to the drainage pool 14 (see FIG. 2).

  In the present embodiment, the first liquid pool 11 is provided so as to be able to store a liquid containing a biomarker to be measured, preferably a biomarker dispersion in which the biomarker is dispersed. However, the first liquid pool 11 is not limited to a biomarker, and may be provided so as to store a liquid containing a specific substance that can be used for analysis or the like. The second liquid pool 12 is provided so as to store a predetermined liquid, for example, a cleaning liquid. The third liquid pool 13 is provided so as to store magnetic beads or a detection substrate. In use, the third liquid pool 13 contains magnetic beads or a detection substrate. The third pool 13 is provided so as to be able to store a liquid containing magnetic beads, preferably a magnetic bead dispersion in which magnetic beads are dispersed. In this case, for example, a method of storing magnetic beads in the third liquid pool 13 and injecting the liquid at the time of use to obtain a liquid containing magnetic beads (preferably a magnetic bead dispersion liquid) may be used. . Details of the biomarkers and magnetic beads will be described later.

[Specific configuration of channel]
FIG. 2 is a plan view showing an example of a specific configuration of the flow path in the sensor cartridge 1 of FIG. 1, and FIG. 3 is a first connection point 41 and a second connection point 42 provided in the flow path of FIG. It is a partial enlarged view which shows the detail of.

  As shown in FIG. 2, the first flow path 31 extends from the first liquid pool 11 toward the reaction field 20 in a straight portion 31a, a bent portion 31b, a straight portion 31c, a bent portion 31d, a straight portion 31e, and a bent portion 31f. And a straight portion 31g in this order. However, the first flow path 31 is not limited to this, and may have a shape different from the above from the viewpoint of miniaturization and layout of the sensor cartridge 1.

  The second flow path 32 has a straight portion 32a and a curved portion 32b in this order from the second liquid pool 12 to the first connection point 41. However, the second flow path 32 is not limited to this, and may have a shape different from the above from the viewpoint of miniaturization and layout of the sensor cartridge 1.

  The third flow path 33 has a straight portion 33a. However, the third flow path 33 is not limited to this, and may have a shape different from the above from the viewpoint of miniaturization and layout of the sensor cartridge 1.

In the present embodiment, at the second connection point 42, the direction from one end 33-1 of the third flow path 33 to the other end 33-2 is from the first liquid pool 11 of the first flow path 31. It has a direction component to the reaction field 20. The width L1 _d2 of the first flow path 31 at the downstream portion 42A of the second connection point 42 is larger than the width L1 _u2 of the first flow path 31 at the upstream portion 42B of the second connection point 42. “Upstream part” means a part immediately before the connection point of interest in the liquid flow direction, and “Downstream part” means a part immediately after the connection point in the liquid flow direction. I do. It is preferable that the third flow path 33 is connected to the first flow path 31 at the second connection point 42 in a direction equivalent to the direction in which the linear portion 31e of the first flow path 31 extends.

As described above, at the second connection point 42, the direction from one end 33-1 of the third flow path 33 to the other end 33-2 is changed from the first liquid pool 11 of the first flow path 31 to the reaction. The first channel 31 is connected to the first channel 31 with a directional component toward the field 20, and the width L1 _d2 of the first channel 31 at the downstream portion 42A is equal to the width L1 _u2 of the first channel 31 at the upstream portion 42B. When the liquid flows from the upstream portion 42B toward the downstream portion 42A in the first flow path 31, it is difficult for the liquid to enter the third flow path 33.

The width L1 _d2 of the first flow path 31 at the downstream portion 42A of the second connection point 42 is the width L1 _u2 of the first flow path 31 and the width of the third flow path 33 at the upstream portion 42B of the second connection point 42. it is preferably larger than the same or該合meter and the sum of L _3. Thus, when the liquid flows from the upstream portion 42B toward the downstream portion 42A in the first flow path 31, the liquid is less likely to enter the third flow path 33.

  Further, the angle between the axial direction of the first flow path 31 and the axial direction of the third flow path 33 at the second connection point 42 is preferably 45 ° or less, more preferably 30 ° or less, and 0 ° (ie, 0 °). , The first flow path 31 is parallel to the third flow path 33 at the second connection point 42). In particular, as shown in FIG. 3B, at the upstream portion 42 </ b> B of the second connection portion 42, the axial direction of the first flow passage 31 is parallel to the axial direction of the linear portion 33 a of the third flow passage 33. Is preferred. As described above, the first flow path 31 is connected in parallel with the third flow path 33 at the second connection point 42, so that the liquid in the first liquid pool 11 or the liquid in the third liquid pool 13 is reacted in the reaction field. When flowing toward 20, the liquid flowing through one flow path is less likely to penetrate into the other flow path.

At the first connection point 41, the direction from one end 32-1 of the second flow path 32 to the other end 32-2 is changed from the first liquid pool 11 of the first flow path 31 to the reaction field 20. It is preferred to have a directional component to At this time, the width L1 _d1 of the first flow path 31 at the downstream portion 41A of the first connection point 41 is equal to the width L1 _u1 of the first flow path 31 at the upstream portion 41B of the first connection point 41 and the width L1 _u1 of the second flow path 32. It is preferable that the width is larger than the width L2. It is more preferable that the second flow path 32 is connected to the first flow path 31 at the first connection point 41 in a direction equivalent to the direction in which the linear portion 31c of the first flow path 31 extends.

Thus, also at the first connection point 41, the direction from one end 32-1 of the second flow path 32 to the other end 32-2 is from the first liquid pool 11 of the first flow path 31. is connected to the first flow path 31 has a direction component of the reaction field 20, further, the width L1 of the first flow path 31 width _{L1 d1} of the first flow path 31 at the downstream portion 41A is in the upstream portion 41B By being larger than _u1, when the liquid flows from the upstream portion 41B toward the downstream portion 41A in the first flow path 31, the liquid is less likely to enter the second flow path 32.

Further, the width L1 _d1 of the first flow path 31 at the downstream portion 41A of the first connection point 41 is the width L1 _u1 of the first flow path 31 and the width of the second flow path 32 at the upstream portion 41B of the first connection point 41. Preferably, it is equal to or greater than the sum of L2. Accordingly, when the liquid flows from the upstream portion 41B toward the downstream portion 41A in the first flow path 31, the liquid is less likely to enter the second flow path 32.

  In addition, the angle between the axial direction of the first flow path 31 and the axial direction of the second flow path 32 at the first connection point 41 is preferably 45 ° or less, more preferably 30 ° or less, and 0 ° (ie, 0 °). , The second flow path 32 is parallel to the first flow path 31 at the first connection point 41). In particular, as shown in FIG. 3B, at the upstream portion 41 </ b> B of the first connection point 41, the axial direction of the second flow path 32 is parallel to the axial direction of the linear portion 31 c of the first flow path 31. Is preferred. As described above, the second flow path 32 is connected in parallel to the first flow path 31 at the first connection point 41, so that the liquid in the first liquid pool 11 or the liquid in the second liquid pool 12 is reacted to the reaction field. When flowing toward 20, the liquid flowing through one flow path is less likely to penetrate into the other flow path.

  The second connection point 42 is preferably provided at a position closer to the reaction field 20 than the first connection point 41 in the first flow path 31 (FIG. 2). Further, the first flow path 31 preferably has a curved portion 31 d on the upstream side of the second connection point 42, and is preferably connected to the non-curved portion of the third flow path 33 at the second connection point 42. In the present embodiment, the curved portion 31d is provided between the first connection portion 41 and the second connection portion 42, and the non-curved portion of the third flow path 33 corresponds to a straight portion 33a. Thus, for example, when a liquid containing magnetic beads or a detection substrate is stored in the third liquid pool 13 and the liquid flows to the reaction field 20 through the third flow path 33, the liquid in the third flow path 33 It is possible to eliminate or reduce the number of times that passes through the curved portion, and it is possible to smoothly supply the magnetic beads or the detection substrate to the reaction field 20.

  The second flow path 32 preferably has a curved portion 32 b upstream of the first connection point 41, and is preferably connected to the non-curved part of the first flow path 31 at the first connection point 41. In the present embodiment, the non-curved portion of the first flow path 31 corresponds to the straight portion 31c. Thereby, when storing the biomarker dispersion liquid in the first liquid pool and flowing the biomarker dispersion liquid to the reaction field 20 via the first flow path 31, the biomarker can be smoothly supplied to the reaction field 20. Becomes possible.

[Specific configuration of magnetic sensor]
The magnetic sensor includes, for example, a substrate, a magnetoresistive element disposed on the substrate, and a protective film formed to cover the magnetoresistive element and the substrate.

  The magnetoresistive element is configured such that a detected resistance value changes according to an input magnetic field.

The protective film has an affinity substance that recognizes the biomarker on the outer surface (hereinafter, also referred to as “first affinity substance”).
The outer surface of the protective film is a surface that comes into contact with the biomarker in the sample. This outer surface is provided with a first affinity substance that specifically binds to the biomarker to be measured. The first affinity substance is, for example, single-stranded DNA or RNA. For example, when the biomarker has single-stranded DNA or RNA, the first affinity substance and the biomarker are specifically bound by hybridization between single-stranded DNA, between RNA, or between single-stranded DNA and RNA. To join. Further, the magnetic beads also include a second affinity substance that specifically binds to the biomarker.

  The configuration of the protective film is not particularly limited as long as it can protect the magnetoresistive element. Materials for the protective film include, for example, oxides such as alumina, silica, titanium oxide, zirconium oxide, indium oxide, tartar oxide, zinc oxide, gallium oxide, and tin oxide; and noble metals such as gold, silver, platinum, rhodium, ruthenium, and palladium. Inorganic materials such as aluminum nitride and silicon nitride, and organic materials such as polyimide.

  When the protective film includes the first affinity substance, the biomarker is immobilized on the outer surface of the protective film via the first affinity substance only when the biomarker to be measured is present in the sample. Further, the magnetic beads bind to the biomarker via the second affinity substance, so that the magnetic beads are fixed on the surface of the protective film.

  Further, the sensor cartridge 1 is provided with an electrode terminal 1D (FIG. 1), and the electrode terminal 1D is electrically connected to the magnetoresistive element.

[Specific configuration of magnetic beads]
The magnetic beads include a second affinity substance that recognizes a site different from the biomarker recognition site of the first affinity substance. The second affinity substance is, for example, streptavidin. For example, when the biomarker has biotin at a site different from single-stranded DNA or RNA, the second affinity substance, streptavidin, specifically binds to biotin. Magnetic beads accumulate on the protective film via a first affinity substance-biomarker-second affinity substance complex.

  In other words, the magnetic beads include a second affinity substance that specifically binds to the biomarker, and binds to the biomarker via the second affinity substance. The magnetic beads may be those to which a second affinity substance has been added by a coating process or the like, or may be composed of the second affinity substance itself.

  The magnetic beads are not particularly limited as long as they are magnetic particles, and include, for example, iron oxide particles. The diameter of the magnetic beads depends on the area of the protective film, but is, for example, preferably from 0.01 μm to 100 μm, more preferably from 0.05 μm to 50 μm, and particularly preferably from 0.1 μm to 5 μm.

[Specific composition of biomarkers]
Examples of the biomarker to be measured or the biomolecule to be detected include, for example, nucleic acids (naturally occurring) such as DNA, mRNA, miRNA, siRNA, and artificial nucleic acids (eg, LNA (Locked Nucleic Acid), BNA (Bridged Nucleic Acid)). And peptides such as ligands, cytokines and hormones; proteins such as receptors, enzymes, antigens and antibodies; cells, viruses, bacteria, fungi and the like.

  Samples (liquids) containing biomolecules to be detected include blood, serum, plasma, urine, puffy coat, saliva, semen, chest exudate, cerebrospinal fluid, tears, sputum, mucus, lymph, ascites, pleural effusion, Amniotic fluid, bladder lavage fluid, bronchoalveolar lavage fluid, cell extract, cell culture supernatant and the like.

  In addition, the biomarker to be measured may be a biomolecule obtained by complexing another biomolecule with a biomolecule to be detected, or a biomolecule obtained by converting a biomolecule to be detected into another biomolecule. Good. For example, a complex of RNA with biotin-terminated DNA by hybridization (hereinafter, may be referred to as “RNA-DNA-biotin complex”) may be used. The addition of biotin to the RNA by complexation allows specific binding to streptavidin.

  In the sensor cartridge 1 configured as described above, different liquids are stored in three liquid pools provided in the sensor cartridge 1 respectively. For example, the first liquid pool 11 stores a biomarker dispersion (for example, a target DNA dispersion), the second liquid pool 12 stores a washing liquid, and the third liquid pool 13 stores a magnetic bead dispersion. As described above, the biomarker dispersion liquid is stored in the first liquid pool 11 by being injected into the first liquid pool 11 using a device such as a syringe.

  Then, with the sensor cartridge 1 set in the magnetic property measuring device (not shown), the floating liquid is applied to the surface of the magnetic sensor as the reaction field 20 in the order of the biomarker dispersion liquid, the washing liquid, and the magnetic bead dispersion liquid. The solution is sent, and the concentration of the biomarker is measured based on the number of magnetic beads.

[How to use the sensor cartridge]
FIGS. 4A to 4F are schematic diagrams for explaining an example of a method of using the sensor cartridge 1 of FIG.

  As shown in FIG. 4A, a first affinity substance 61 such as DNA or an antibody is fixed on the surface of the magnetic sensor that is the reaction field 20, that is, on the outer surface of the protective film.

  First, a biomarker dispersion is supplied from the first liquid pool 11 to the reaction field 20 via the first flow path 31 (FIG. 4B), and a biomarker 62 is attached to the first affinity substance 61. It is fixed (FIG. 4C).

  Here, as shown in the above-mentioned prior art, when the connection point between the first flow path in which the biomarker dispersion flows and the third flow path in which the magnetic beads dispersion flows has a T-shape, that is, the third flow path When the biomarker is connected vertically to the first channel, the biomarker flows into the third channel and remains in the third channel when the biomarker dispersion liquid flows through the first channel, and the biomarker is removed. It may react with magnetic beads. Further, since the width of the first channel is constant, there is a high possibility that the biomarker flows from the first channel into the third channel and remains in the third channel.

On the other hand, in the present embodiment, at the second connection point 42, the direction from one end 33-1 of the third flow path 33 to the other end 33-2 is the first liquid pool of the first flow path 31. 11 to the reaction field 20, and the width L1 _d2 of the first flow path 31 in the downstream portion 42A of the second connection point 42 is the first component in the upstream portion 42B of the second connection point 42. Since the width of the flow path 31 is larger than the width L1 _u2 , the flow of the biomarker dispersion liquid into the first flow path 31 makes it difficult for the biomarker to flow into the third flow path 33.

  Next, the washing liquid is supplied from the second liquid pool 12 to the reaction field 20 via the second flow path 32 and the first flow path 31 to remove the biomarker remaining in the first flow path 31 and to remove the first affinity. Excess biomarkers 62 not captured by the substance 61 are removed from the magnetic sensor or its vicinity (FIG. 4D).

Also in this step, the direction from one end 32-1 of the second flow path 32 to the other end 32-2 of the first connection point 41 is shifted from the first liquid pool 11 of the first flow path 31. It has a directional component toward the reaction field 20, and the width L1 _d1 of the first flow path 31 at the downstream part 41A of the first connection point 41 is equal to the width L1 of the first flow path 31 at the upstream part 41B of the first connection point 41. _Since it is larger than _u1, it is difficult for the biomarker 62 to flow into the second flow path 32 when the biomarker dispersion liquid flows into the first flow path 31, and the mixing of the biomarker 62 into the cleaning liquid is suppressed. Therefore, the surface of the first flow path 31 and the surface of the magnetic sensor, which is the reaction field 20, can be sufficiently washed with the original washing liquid in which the biomarker 62 is not mixed.

  Thereafter, a magnetic bead dispersion is supplied from the third liquid pool 13 to the reaction field 20 via the third flow path 33 and the first flow path 31 (FIG. 4E), and the magnetic beads 63 are supplied to the biomarker 62. Affix and fix (FIG. 4 (f)). Since the first flow path 31 is washed, the mixing of the magnetic bead dispersion liquid and the biomarker dispersion liquid is suppressed on the upstream side of the reaction field. The magnetic beads 63 used in the present embodiment are covered with this specific protein, and when the magnetic beads 63 are supplied on the magnetic sensor to which the biomarkers 62 are fixed, the magnetic beads 63 bind to the biomarkers 62. Stay with the biomarker 62 on the magnetic sensor.

  In this state, an external magnetic field is applied near the magnetic sensor. Since the resistance value of the magnetic sensor changes in accordance with the number of magnetic beads 63 on the magnetic sensor, the number of magnetic beads on the magnetic sensor is measured by the amount of change in resistance, and the biometric value is determined from the measured value of the magnetic beads. The concentration of the marker 62 is measured.

As described above, according to the present embodiment, the direction from one end 33-1 to the other end 33-2 of the third flow path 33 at the second connection point 42 is the same as that of the first flow path 31. Has a directional component from the first liquid pool 11 to the reaction field 20, and the width L1 _d2 of the first flow path 31 at the downstream portion 42A of the second connection point 42 is upstream of the second connection point 42. Since the biomarker dispersion liquid flows from the upstream part 42B toward the downstream part 42A in the first flow path 31 because the width of the biomarker dispersion liquid is larger than the width L1 _{u2 of} the first flow path 31 in the part 42B, It is difficult to enter the road 33. This can prevent the magnetic beads 63 that have reacted with the biomarker 62 from being supplied to the surface of the magnetic sensor that is the reaction field 20, and can improve the measurement accuracy of the biomarker 62. Thereby, for example, the concentration of a biomarker whose concentration changes depending on the disease state, which is contained in a sample derived from a living body such as blood, plasma, saliva and sweat, is measured simply, quickly and with high accuracy, and the measurement result is obtained. Can be used for diagnosis of humans and the like.

[Modification of Sensor Cartridge and How to Use It]
FIGS. 5A to 5D are schematic diagrams for explaining a modified example of the sensor cartridge 1 of FIG. 1 and a method of using the modified example. This modification is different from the above embodiment in that a substrate is provided instead of a magnetic sensor, a liquid containing a biomarker having an enzyme is used, and a liquid containing a detection substrate is used instead of a magnetic bead dispersion. The substrate provided in place of the magnetic sensor has a first affinity substance on its surface, and the substrate surface becomes a reaction field.

  In a modified example of the sensor cartridge 1, for example, a biomarker dispersion liquid in which a biomarker having an enzyme is dispersed is stored in the first liquid pool 11, and a cleaning liquid is stored in the second liquid pool 12, respectively. The third liquid pool 13 stores a liquid containing a detection substrate, preferably a detection substrate dispersion in which a detection substrate that reacts with the specific enzyme is dispersed.

  Examples of the above specific enzyme include alkaline phosphatase and horseradish peroxidase. The biomarker having an enzyme may be, for example, a biomarker in which an enzyme is directly bound to a biomarker, or a biomarker in which streptavidin added to the enzyme and biotin added to the biomarker are bound.

  Examples of the detection substrate include a chromogenic substrate, a chemiluminescent substrate, and a chemiluminescent substrate. From the viewpoint of stable color development, a chromogenic substrate is preferable. Examples of the chromogenic substrate include, for example, when using alkaline phosphatase as the enzyme, paranitrophenyl phosphate, and when using horseradish peroxidase as the enzyme, 3,3 ′, 5,5′-tetramethyl Benzidine.

  An example of how to use the modified example of the sensor cartridge will be described. As shown in FIG. 5A, a first affinity substance 61 such as DNA or antibody is immobilized on the surface of the substrate which is the reaction field 20.

  First, a dispersion containing a biomarker 62 having an enzyme 71 is supplied from the first liquid pool 11 to the reaction field 20 via the first flow path 31 (FIG. 5B). Then, a biomarker 62 having an enzyme 71 is attached and fixed (FIG. 5C). Next, a cleaning liquid is supplied from the second liquid pool 12 to the reaction field 20 via the second flow path 32 and the first flow path 31, and the biomarker 62 having the enzyme 71 remaining in the first flow path 31 is removed. At the same time, excess biomarkers 62 not captured by the first affinity substance 61 are removed from the substrate or its vicinity (FIG. 5D).

  Thereafter, the detection substrate dispersion is supplied from the third liquid pool 13 to the reaction field 20 via the third flow path 33 (FIG. 5E), and the detection substrate 72 is reacted with the enzyme 71 (FIG. 5F). ). For example, when the detection substrate dispersion is a chromogenic substrate dispersion, the amount of coloring (shading of the color) of the detection substrate 72 is determined according to the amount of the enzyme 71. The value of light absorption at a specific wavelength is measured from the amount of coloring (color density) of the detection substrate 72. That is, absorbance detection (colorimetric analysis) is performed, and the concentration of the biomarker 62 is measured from the value of light absorption at a specific wavelength on the substrate.

Also in the present modified example, the direction from one end 33-1 of the third flow path 33 to the other end 33-2 of the third flow path 33 at the second connection point 42 is the first liquid pool 11 of the first flow path 31. And the width L1 _d2 of the first flow path 31 at the downstream portion 42A of the second connection point 42 is different from the first flow rate at the upstream portion 42B of the second connection point 42. Since the width of the passage 31 is larger than the width L1 _u2, when the biomarker dispersion flows from the upstream portion 42B toward the downstream portion 42A in the first channel 31, the biomarker dispersion does not easily enter the third channel 33. . If the enzyme 71 and the detection substrate 72 are mixed and reacted upstream of the reaction field 20, the detection substrate 72 that has reacted with the “biomarker 62 having the enzyme 71” that does not bind to the first affinity substance 61 As a result, the amount of light by the detection substrate 72 changes, and the accuracy is greatly reduced. On the other hand, according to the present modification, the detection substrate 72 that has reacted with the biomarker 62 having the enzyme 71 can be prevented from being supplied to the surface of the substrate that is the reaction field 20, and the biomarker 62 having the enzyme 71 can be prevented. Measurement accuracy can be improved.

  As described above, the embodiments of the present invention have been described in detail. However, the present invention is not limited to the above embodiments, and various modifications and changes may be made within the scope of the present invention described in the appended claims. Is possible.

  For example, in the above embodiment, the sensor cartridge 1 has three liquid pools and three flow paths corresponding to the liquid pools. However, the present invention is not limited to this, and four or more liquid pools and four or more May be provided. Also in this case, it is possible to prevent liquids from being mixed in each flow path.

  Further, in the above embodiment, the first liquid pool 11 stores the liquid containing the biomarker 62, and the second liquid pool 12 stores the cleaning liquid. However, the present invention is not limited to this, and the first liquid pool 11 stores the cleaning liquid. The second liquid pool 12 may store liquid containing the biomarker 62.

  The sensor cartridge of the present invention can be applied to, for example, cancer diagnosis, diagnosis by cancer type, diagnosis of cancer progression, detection of influenza virus, identification of influenza virus type, observation of influenza pathology, and the like. .

DESCRIPTION OF SYMBOLS 1 Sensor cartridge 1A Base plate 1A-1 Upper surface 1B Substrate 1B-1 Upper surface 1C Cover plate 1C-1 Upper surface 1C-2 Lower surface 1D Electrode terminal 11 1st liquid pool 12 2nd liquid pool 13 3rd liquid pool 14 Drainage pool 20 Reaction Field 21 hole 31 First flow path 31a Straight part 31b Curved part 31c Straight part 31d Curved part 31e Straight part 31f Curved part 31g Straight part 32 Second flow path 32a Straight part 32b Curved part 32-1 One end 32-2 The other end 33 Third flow path 33-1 One end 33-2 The other end 34 Fourth flow path 41 First connection point 41A Downstream section 41B Upstream section 42 Second connection point 42A Downstream section 42B Upstream section 51 cap member 52 cap member 53 cap member 61 first affinity substance 62 biomarker 3 magnetic beads 71 enzyme 72 detection substrate

Claims (9)

A first liquid pool, a second liquid pool, and a third liquid pool;
A reaction field,
A first flow path connected to the first liquid pool and configured to be able to transport liquid from the first liquid pool onto the reaction field;
A second flow path having one end connected to the second liquid pool and the other end connected to the first flow path at a first connection point;
A third flow path having one end connected to the third liquid pool and the other end connected to the first flow path at a second connection point;
At the second connection point, the direction from the one end of the third flow path to the other end has a directional component from the first liquid pool of the first flow path to the reaction field. And
A sensor cartridge in which a width of the first flow path at a downstream part of the second connection point is larger than a width of the first flow path at an upstream part of the second connection point.   The width of the first flow path at the downstream part of the second connection point is equal to or the same as the sum of the width of the first flow path and the width of the third flow path at the upstream part of the second connection point. The sensor cartridge according to claim 1, wherein the sensor cartridge is larger than the sensor cartridge. At the first connection point, the direction from the one end of the second flow path to the other end has a directional component from the first liquid pool of the first flow path to the reaction field. And
3. The sensor cartridge according to claim 1, wherein a width of the first flow path at a downstream part of the first connection point is larger than a width of the first flow path at an upstream part of the first connection point. 4.   The width of the first flow path at the downstream part of the first connection point is equal to or the same as the sum of the width of the first flow path and the width of the second flow path at the upstream part of the first connection point. The sensor cartridge according to claim 3, wherein the sensor cartridge is larger than the sensor cartridge. The second connection point is provided at a position closer to the reaction field than the first connection point in the first flow path,
5. The first flow path according to claim 1, wherein the first flow path has a curved portion on an upstream side of the second connection point, and is connected to a non-curved portion of the third flow path at the second connection point. 6. The sensor cartridge according to claim 1.   The sensor according to claim 5, wherein the second flow path has a curved portion on an upstream side of a first connection point, and is connected to a non-curved portion of the first flow path at the first connection point. cartridge.   The sensor cartridge according to claim 1, wherein the first flow path is parallel to the third flow path at the second connection point.   The sensor cartridge according to claim 1, wherein the second flow path is parallel to the first flow path at the first connection location.   The sensor cartridge according to claim 1, wherein the third liquid pool stores magnetic beads or a detection substrate.

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