JP2016114409A - Substrate for sample analysis, sample analysis device, sample analysis system, and program for sample analysis system - Google Patents

Abstract

Provided are a sample analysis substrate and a sample analysis system that can cope with an analysis method in which a component in a specimen is analyzed through more complicated reaction steps.
A sample analysis substrate is a sample analysis substrate that transfers liquid by rotational movement, and includes a substrate 100 ′ having a plate shape having a rotation axis and a predetermined thickness, and a position within the substrate. A first chamber 102 having a first space for holding the liquid, a second chamber 103 having a second space for holding the liquid discharged from the first chamber in the substrate, and the substrate. And a first flow path 112 having a path connecting the first chamber and the second chamber, and at least a part of the liquid disposed in the second space of the second chamber and discharged into the second chamber Liquid absorbing members 121, 122 capable of absorbing water.
[Selection] Figure 3B

Description

  The present application relates to a sample analysis substrate, a sample analysis apparatus, a sample analysis system, and a sample analysis system program.

  Conventionally, a technique using a sample analysis substrate for analyzing a specific component in a specimen such as urine or blood is known. For example, Patent Document 1 uses a disk-shaped sample analysis substrate on which a channel, a chamber, and the like are formed, and rotates the sample analysis substrate to transfer, distribute, and mix the components in the sample solution. The technology which performs analysis etc. of this is disclosed.

JP 7-500910 Gazette

  Analysis of a specific component in a specimen includes an analysis method using complicated reaction steps using an enzyme reaction, an immune reaction, or the like. There has been a demand for a technique capable of performing an analysis method through such a complicated reaction step in a sample analysis substrate.

  Non-limiting exemplary embodiments of the present application provide a sample analysis substrate, a sample analysis apparatus, a sample analysis system, and a sample analysis that can be applied to an analysis method in which components in a specimen are analyzed through more complicated reaction steps. Provide system programs.

  A sample analysis substrate according to the present disclosure is a sample analysis substrate that transfers a liquid by a rotational motion, the substrate having a plate shape having a rotation axis and a predetermined thickness, and located in the substrate. A first chamber having a first space for holding a liquid, a second chamber having a second space for holding the liquid discharged from the first chamber in the substrate, and the substrate A first channel having a path connecting the first chamber and the second chamber, and a liquid disposed in the second space of the second chamber and discharged into the second chamber A liquid absorbing member capable of absorbing at least a part of the liquid absorbing member.

  According to the sample analysis substrate, the sample analysis apparatus, the sample analysis system, and the sample analysis system program according to one aspect of the present application, it is possible to cope with an analysis method in which components in a specimen are analyzed through complicated reaction steps.

FIG. 1 is an example of a schematic diagram illustrating a sandwich immunoassay method using magnetic particles. FIG. 2 is an example of a schematic diagram illustrating the configuration of the sample analysis system of the embodiment. FIG. 3A is an example of an exploded perspective view of a sample analysis substrate. FIG. 3B is an example of a plan view of the sample analysis substrate. FIG. 3C is an example showing the thickness of each chamber and flow path in the cross section of the thick broken line portion in FIG. 3B. FIG. 3D is an example showing a positional relationship from the rotation axis on the sample analysis substrate in the reaction chamber, the B / F separation chamber, the recovery chamber, the second path, and the fourth path. FIG. 4 is an example of a flowchart for explaining the operation of the sample analysis system. FIG. 5 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 6 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 7 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 8 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 9 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 10 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 11 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 12 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 13 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 14 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 15 is an example of a diagram schematically showing the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system. FIG. 16A is an example of a schematic diagram illustrating a state in which the reaction liquid or the cleaning liquid remains in the first sub chamber when there is no liquid absorbing member. FIG. 16B is an example of a schematic diagram illustrating a state in which the reaction liquid or the cleaning liquid in the second sub chamber flows back to the B flow path when there is no liquid absorbing member. FIG. 17 is another example of a plan view of the sample analysis substrate. FIG. 18 is another example of a plan view of the sample analysis substrate.

  As a method for analyzing a component of a specimen such as urine or blood, a binding reaction between an analyte as an analysis target and a ligand that specifically binds to the analyte may be used. Examples of such analysis methods include immunoassay methods and genetic diagnosis methods.

  As an example of the immunoassay method, there are a competitive method and a non-competitive method (sandwich immunoassay method). An example of a gene diagnosis method is a gene detection method by hybridization. As an example of these analysis methods, a sandwich immunoassay method using magnetic particles (sometimes referred to as “magnetic beads”, “magnetic particles” or “magnetic beads”) will be specifically described.

  As shown in FIG. 1, first, an antigen-antibody reaction between a primary antibody 304 immobilized on the surface of a magnetic particle 302 (hereinafter referred to as “magnetic particle-immobilized antibody 305”) and an antigen 306 as a measurement object. And combine with. Next, the secondary antibody to which the labeling substance 307 is bound (hereinafter referred to as “labeled antibody 308”) and the antigen 306 are bound by an antigen-antibody reaction. As a result, a complex 310 in which the magnetic particle-immobilized antibody 305 and the labeled antibody 308 are bound to the antigen 306 is obtained.

  A signal based on the labeling substance 307 of the labeled antibody 308 bound to the complex 310 is detected, and the antigen concentration is measured according to the amount of the detected signal. Examples of the labeling substance 307 include enzymes (for example, peroxidase, alkaline phosphatase, luciferase, etc.), chemiluminescent substances, electrochemiluminescent substances, fluorescent substances, etc., and dyes corresponding to the respective labeling substances 307, Signals such as luminescence and fluorescence are detected.

  In this series of reactions, in order to obtain the complex 310 as a reactant, an unreacted substance in the sample, a substance adsorbed non-specifically to magnetic particles, etc., a labeled antibody 308 that was not involved in the formation of the complex 310 It is necessary to separate unreacted substances that are equal to each other. This separation is called B / F separation (Bound / Free Separation). Similarly, the B / F separation step is necessary in the immunoassay method by the competitive method and the gene detection method by hybridization.

  As described above, the sandwich immunoassay method using magnetic particles has been described as an example, but B / F separation is performed by immunoassay or hybridization using competitive or non-competitive methods regardless of the presence or absence of magnetic particles. Necessary for gene detection. When magnetic particles are not used, for example, a ligand immobilized on a solid phase composed of a material such as polystyrene or polycarbonate (immobilization by physical adsorption), a metal substrate surface composed of gold or the like (for example, Examples include the case of using a ligand that is self-assembled monolayer (immobilized using SAM: self-assembled monolayer).

  In order to sufficiently perform the B / F separation, it is preferable to wash the magnetic particles containing the composite 310 multiple times with a washing liquid. Specifically, first, in the reaction solution containing the complex 310, the unreacted antigen 306, the labeled antibody 308, and the like, only the reaction solution is removed while the complex 310 containing the magnetic particles is captured by the magnet. . Thereafter, a cleaning liquid is added to clean the composite 310, and the cleaning liquid is removed. By repeating this washing a plurality of times, B / F separation in which unreacted substances are sufficiently removed can be achieved. This point is also described in an example using magnetic particles, but this is true for sandwich-type assays in general, regardless of whether magnetic particles are used.

  Conventionally, such an operation of performing multiple times of washing has been realized manually by an operator using an analytical instrument or a large analytical instrument having a complicated mechanism. For this reason, there has been a demand for a technique for performing cleaning more than once more easily.

The inventors of the present application have studied in detail a technique that enables a plurality of cleaning steps using a sample analysis substrate as disclosed in Patent Document 1, and have developed a novel sample analysis substrate, a sample analysis device, A sample analysis system and a program for the sample analysis system were conceived. A sample analysis substrate, a sample analysis device, a sample analysis system, and a sample analysis system program according to an aspect of the present application are as follows.
[Item 1]
A sample analysis substrate for transferring a liquid by a rotational motion, the substrate having a plate shape having a rotation axis and a predetermined thickness, and a first space located in the substrate for holding the liquid A first chamber having a second space for holding the liquid discharged from the first chamber in the substrate, the first chamber being located in the substrate, A first flow path having a path connecting the second chamber and the second flow path disposed in the second space of the second chamber and absorbing at least part of the liquid discharged into the second chamber. A sample analysis substrate comprising a liquid absorbing member that can be formed. According to this configuration, since the liquid absorbing member can absorb the liquid transferred to the second chamber, the liquid in the second chamber flows back to the first chamber even when the sample analysis substrate stops at an unintended angle. Can be suppressed.
[Item 2]
The liquid absorbing member has a third space having a predetermined volume between the liquid absorbing member and a connection portion between the first flow path and the second chamber in the second space. Item 2. The sample analysis substrate according to Item 1, which is arranged. According to this configuration, since the liquid transferred from the first flow path can move into the second chamber through the third space, the liquid is transferred even when the liquid is transferred from the first chamber a plurality of times. Can be securely held in the second chamber.
[Item 3]
Item 3. The sample analysis substrate according to Item 1 or 2, wherein the first channel can fill the first channel with a liquid held in the first space by a capillary phenomenon.
[Item 4]
The second chamber includes a first sub chamber, a second sub chamber, and a connecting portion that connects the first sub chamber and the second sub chamber,
4. The sample analysis substrate according to any one of items 1 to 3, wherein the second sub chamber is located farther from the rotation axis than the first sub chamber.
[Item 5]
Item 5. The sample analysis substrate according to Item 4, wherein the liquid absorbing member is disposed in the first sub chamber. According to this structure, when a liquid remains in a 1st subchamber, it can suppress that a liquid flows backward to a 1st flow path.
[Item 6]
Item 5. The sample analysis substrate according to Item 4, wherein the liquid absorbing member is disposed in the second sub chamber. According to this structure, when a liquid remains in a 2nd subchamber, it can suppress that a liquid flows backward to a 1st flow path.
[Item 7]
A capillary chamber having a third chamber located in the substrate and having a third space for holding a liquid; and a path located in the substrate and connecting the third chamber and the first chamber; And a second flow path that can be filled with a liquid held in the third space due to a phenomenon, the second flow path has a first opening and a second opening, and the first opening and the first flow 2 openings are connected to the third chamber and the first chamber, respectively, the first opening is located closer to the rotation axis than the second opening, and the first space includes the first area and the second area. The first region includes a portion connected to the first opening and extending from the first opening toward a side farther from the rotation axis, and the second region rotates more than the first opening. The first region at a position far from the axis; Being connected to the portion of extension, said third space of the third chamber is greater than the volume of the second flow path, the substrate for sample analysis as described in any one of 1 to 6. According to this configuration, when the liquid held in the third chamber is moved to the first chamber in multiple steps and further moved to the second chamber, the sample analysis substrate is stopped at various angular positions. However, since the liquid absorbing member in the second chamber absorbs at least a part of the liquid in the second chamber, it is possible to prevent the liquid in the second chamber from flowing backward from the first flow path to the first chamber. it can.
[Item 8]
8. The sample analysis substrate according to any one of items 1 to 7, further comprising a magnet positioned in proximity to the first chamber.
[Item 9]
The sample analysis substrate according to any one of items 1 to 8 and the sample analysis substrate are rotated around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity. A rotation angle detector for detecting a rotation angle of a rotation shaft of the motor, a driver for controlling a rotation angle at the time of rotation and stop of the motor based on a detection result of the rotation angle detector, an arithmetic unit, and a memory And a controller stored in the memory and configured to be executable by the computing unit, and based on the program, a controller for controlling operations of the motor, the rotation angle detector, the origin detector, and the driver A sample analysis system comprising: a sample analysis system comprising: a sample analysis substrate in which the first chamber is filled with a liquid; (A) When the sample analysis substrate is rotated, at least a part of the liquid in the first chamber is transferred to the second chamber via the first flow path. Let the sample analysis system.
[Item 10]
A motor for rotating the sample analysis substrate according to any one of items 1 to 8 around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity. Rotation angle detector that detects the rotation angle of the rotation shaft, a driver that controls the rotation angle of the motor at the time of rotation and stop based on the detection result of the rotation angle detector, and an arithmetic unit, memory, and memory The sample analyzer includes a program configured to be executable by the computing unit, and includes a controller that controls operations of the motor, the rotation angle detector, the origin detector, and the driver based on the program And when the sample analysis substrate in which the liquid is filled in the first chamber is loaded in the sample analyzer, (a) Fee analytical substrate is rotated, said at least a portion of the liquid in the first chamber, the sample analyzing apparatus for transferring via the first passage to the second chamber.
[Item 11]
The sample analysis substrate according to any one of items 1 to 8 and the sample analysis substrate are rotated around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity. A rotation angle detector that detects the rotation angle of the rotation shaft of the motor, a driver that controls the rotation angle of the motor and the rotation angle when stopped based on the detection result of the rotation angle detector, and an arithmetic unit, A memory and a program stored in the memory and configured to be executable by the computing unit, and based on the program, control for controlling operations of the motor, the rotation angle detector, the origin detector, and the driver A sample analysis system program comprising a sample analysis device having a vessel, wherein the program is for sample analysis in which the first chamber is filled with a liquid When the plate is loaded in the sample analyzer, (a) by rotating the sample analysis substrate, at least a part of the liquid in the first chamber is allowed to pass through the first channel. Sample analysis system program to be transferred to two chambers.

  Hereinafter, a sample analysis substrate, a sample analysis device, a sample analysis system, and a sample analysis system program according to the present embodiment will be described in detail with reference to the drawings. The sample analysis substrate, sample analysis apparatus, sample analysis system, and sample analysis system program of this embodiment weigh a certain amount of liquid held in one chamber, divide it into multiple chambers, and transfer it to another chamber. Can be transported. In the embodiment, it is described that the liquid is a cleaning liquid, but the liquid is not limited to the cleaning liquid, and may be various liquids used for sample analysis.

  FIG. 2 is a schematic diagram showing the overall configuration of the sample analysis system 501. The sample analysis system 501 includes a sample analysis substrate 100 and a sample analysis device 200.

(Configuration of sample analyzer 200)
The sample analyzer 200 includes a motor 201, an origin detector 203, an angle detector 204, a controller 205, a driver 206, and an optical measuring device 207.

  The motor 201 has a rotation axis A inclined from the direction of gravity at an angle θ of greater than 0 ° and 90 ° or less with respect to the turntable 201a and the direction of gravity, and the sample analysis substrate 100 placed on the turntable 201a. Rotate around the rotation axis A. Since the rotation axis A is inclined, gravity can be used in addition to centrifugal force due to rotation for transferring the solution on the sample analysis substrate 100. For example, the motor 201 may be a DC motor, a brushless motor, or the like.

  The origin detector 203 detects the origin of the sample analysis substrate 100 attached to the motor 201. For example, the origin detector 203 includes a light source and a light receiving element, and is arranged such that the sample analysis substrate 100 is positioned between the light source and the light receiving element. The sample analysis substrate 100 has a light shielding portion that shields light at a specific position. When the sample analysis substrate 100 is rotated by the motor 201, the light emitted from the light source of the origin detector 203 is blocked by the light shielding unit at a specific angle of the sample analysis substrate 100. At this time, the light receiving element outputs a change in the detected light. As a result, the origin position of the sample analysis substrate 100 (the angular position serving as the reference of the sample analysis substrate 100) is detected. The origin detector 203 may have other configurations. For example, the sample analysis substrate 100 may include an origin detection magnet, and the origin detector 203 may be a magnetic detection element that detects the magnetism of the magnet. Moreover, you may use the magnet for catching the magnetic particle mentioned later for origin detection. When the sample analysis substrate 100 can be attached to the turntable 201a only at a specific angle, the origin detector 203 may not be provided.

  The angle detector 204 detects the angle of the rotation axis A of the motor 201. For example, the angle detector 204 may be a rotary encoder attached to the rotation axis A. When the motor 201 is a brushless motor, the angle detector 204 may be a Hall element provided in the brushless motor.

  The driver 206 rotates the motor 201. Specifically, the sample analysis substrate 100 is rotated clockwise or counterclockwise based on a command from the controller. Further, based on the detection results of the angle detector 204 and the origin detector 203 and the command from the controller, the sample analysis substrate 100 is stopped from swinging and rotating.

  The optical measuring device 207 detects a signal (eg, dye, luminescence, fluorescence, etc.) corresponding to the labeling substance 307 of the labeled antibody 308 bound to the complex 310 (FIG. 1) held on the sample analysis substrate 100.

  The controller 205 includes a calculator, a memory, and a program stored in the memory and configured to be executable by the calculator. The controller 205 controls operations of the motor 201, the origin detector 203, the angle detector 204, the driver 206, and the optical measuring device 207 based on the program.

(Sample analysis substrate 100)
FIG. 3A is an exploded perspective view of the sample analysis substrate 100. The sample analysis substrate 100 includes a rotating shaft 110 and a plate-shaped substrate 100 ′ having a predetermined thickness along the rotating shaft. In the present embodiment, the substrate 100 ′ of the sample analysis substrate 100 has a circular shape, but may have a polygonal shape or an elliptical shape. In the present embodiment, the substrate 100 ′ is described as having two main surfaces parallel to each other and a predetermined thickness (distance between the two main surfaces) being the same plate shape. The two major surfaces need not be parallel. For example, a part of two main surfaces may be parallel, or may be non-parallel as a whole. In the present embodiment, the substrate 100 ′ of the sample analysis substrate 100 is composed of a base substrate 100a and a cover substrate 100b.

  FIG. 3B is a plan view of the base substrate 100a. As shown in FIG. 3B, the sample analysis substrate 100 includes a cleaning liquid chamber 101 (third chamber), a B / F separation chamber (first chamber) 102, a recovery chamber 103 ( A second chamber), a storage chamber 104, and a reaction chamber 105. The shape of each chamber is not limited as long as it is not specifically mentioned below, and may have any shape. Each chamber generally has a space defined by an upper surface and a lower surface parallel to the two major surfaces of the substrate 100 ′ and four side surfaces located therebetween. Two adjacent surfaces of the upper surface, the lower surface, and the side surface may not be separated by a clear ridge line. For example, the shape of each chamber may be a flat sphere or a spheroid. The sample analysis substrate 100 includes an A channel 111 (second channel), a B channel (first channel) 112, a C channel 113, and a D channel 114, which are located in the substrate 100 ′. It has further. The A channel 111 has a path connecting the cleaning liquid chamber 101 and the B / F separation chamber 102. Similarly, the B channel 112 has a path connecting the B / F separation chamber 102 and the recovery chamber 103. The C flow path 113 has a path connecting the storage chamber 104 and the cleaning liquid chamber 101. The D channel 114 has a path connecting the reaction chamber 105 and the B / F separation chamber 102. Alphabets A to D attached to the head of the names of the channels are simply symbols for distinguishing a plurality of channels, and do not mean the shape of the channels or the characteristics of the channels.

  The sample analysis substrate 100 further includes liquid absorbing members 121 and 122 arranged in the space of the recovery chamber 103.

  The spaces of the cleaning liquid chamber 101, the B / F separation chamber 102, the recovery chamber 103, the storage chamber 104, and the reaction chamber 105 are formed in the base substrate 100a, and the base substrate 100a is covered with the cover substrate 100b. The upper part or the lower part is formed. That is, these spaces are defined by the inner surface of the substrate 100 '. The A channel 111, the B channel 112, the C channel 113, and the D channel 114 are also formed in the base substrate 100a, and by covering the base substrate 100a with the cover substrate 100b, A lower part is formed. In this embodiment, the base substrate 100a and the cover substrate 100b are used as the upper surface and the lower surface, respectively. The substrate 100 ′ can be made of a resin such as acrylic, polycarbonate, or polystyrene.

  As described with reference to FIG. 1, the reaction chamber 105 is a reaction field in which a magnetic particle-immobilized antibody 305, a specimen containing an antigen 306, and a labeled antibody 308 are reacted simultaneously to form a complex 310. is there.

  In this embodiment mode, the structure in which the reaction chamber 105 is separately provided as a reaction field for forming the complex 310 is shown. However, the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308 are supplied to the reaction chamber 107. Various means can be used for the transfer of the above.

  For example, a means is used in which a mixed solution in which a specimen containing the magnetic particle-immobilized antibody 305, the antigen 306, and the labeled antibody 308 is mixed in advance into the sample analysis substrate 100 to form a complex in the reaction chamber 105. obtain.

  For example, a magnetic particle-immobilized antibody 305, a chamber holding each of the specimen containing the antigen 306 and the labeled antibody 308, and a flow path (for example, a capillary flow path) connecting each chamber and the reaction chamber 105, The sample containing the magnetic particle-immobilized antibody 305, the antigen 306, and the labeled antibody 308 can be transferred from the chamber to the reaction chamber 105 and mixed in the reaction chamber 105 to form a complex 310.

  The solution containing the composite 310 is transferred to the B / F separation chamber 102 via the D channel 114.

  The storage chamber 104 stores a cleaning liquid used for cleaning at the time of B / F separation. As will be described in detail below, in the sample analysis system of the present embodiment, the complex 310 can be washed multiple times during the B / F separation. Therefore, the storage chamber 104 can hold a total volume of cleaning liquid corresponding to the number of cleanings.

  The cleaning liquid chamber 101 holds all the cleaning liquid stored in the storage chamber 104. Thereafter, in order to clean the composite 310 in the B / F separation chamber 102, a part of the cleaning liquid is transferred to the B / F separation chamber 102 and the rest is retained. The amount of the cleaning liquid used for one cleaning is weighed by the A channel 111 as described below. For this reason, the cleaning liquid chamber 101 has a volume larger than the A channel 111 and has a volume equal to or larger than the total amount of the cleaning liquid corresponding to the number of times of cleaning (for example, twice or more than the A channel 111 in the case of two cleanings). 3 volumes or more than 3 times the volume of the A channel 111 in the case of washing three times.

  The space (first space) of the cleaning liquid chamber 101 includes a first region 101a and a second region 101b. The first region 101a is connected to a first opening 111c of the A channel 111 described below. The first region 101a includes a portion extending from the first opening 111c toward the side farther from the rotation shaft 110. Thereby, the cleaning liquid located in the first region 101 a can be transferred to the B / F separation chamber 102 via the A channel 111. The second region 101b is connected to the first region 101a) at a position farther from the rotation shaft 110 than the first opening 111c. That is, the second region 101b includes a portion located farther from the rotation axis 110 than the first region 101a, and the second region 101b portion connected to the first region 101a is further away from the rotation shaft 110 than the first region 101a. It is a part located far away. Moreover, it is possible to hold the cleaning liquid in excess in the amount used for one cleaning in the second region 101b. In order to transfer the cleaning liquid from the storage chamber 104 to the cleaning liquid chamber 101, at least a part of the second region 101 b is located farther from the rotating shaft 110 than the storage chamber 104.

  As will be described in detail below, the first region 101a includes a connection portion 101c connected to the first opening 111c in order to smoothly move the cleaning liquid to the A channel 111. The connecting portion 101c is at least a part of the first region 101a and extends from the first opening 111c to the side farther from the rotating shaft 110. The connecting part 101c can suck the cleaning liquid held in the cleaning liquid chamber 101 by capillary action and hold it in the connecting part 101c. This makes it possible to move the cleaning liquid to the A channel 111 more reliably.

  The B / F separation chamber 102 is a place where B / F separation of the solution containing the composite 310 is performed. For B / F separation, the sample analysis substrate 100 includes a magnet 116 disposed in the substrate 100 ′. The magnet 116 is positioned in the vicinity of the space of the B / F separation chamber 102 in the sample analysis substrate 100. More specifically, the magnet 116 is disposed in the vicinity of the side surface 102 s located farthest from the rotation axis among the plurality of side surfaces of the B / F separation chamber 102. However, the magnet 116 in the sample analysis substrate 100 may be disposed at a position close to the upper surface or the lower surface other than the side surface 102 s of the B / F separation chamber 102. That is, the position is not particularly limited as long as the magnetic particles can be captured on the wall surface of the B / F separation chamber 102 by the magnet 116. The magnet 116 may be configured to be removable according to B / F separation, may be attached to the substrate 100 ′ in a non-detachable manner, or may be configured to be provided on the sample analyzer 200 side. . When the magnet 116 is provided on the sample analyzer 200 side, for example, the turntable 201a of the sample analyzer 200 may include a magnet unit including the magnet 116. In this case, when the user arranges the sample analysis substrate 100 at a predetermined position of the turntable 201 a (magnet unit), the magnet 116 is arranged at a position where the magnetic particles can be captured on the wall surface of the B / F separation chamber 102. Further, for example, the sample analyzer 200 may be provided with a drive mechanism that can attach and detach the magnet 116 in the sample analysis substrate 100 in accordance with B / F separation.

  When the reaction solution is transferred to the B / F separation chamber 102 via the D channel 114, the complex 310 in the reaction solution and the unreacted magnetic particle-immobilized antibody 305 (hereinafter, both of them are referred to) The magnetic particles 311 are simply captured on the side of the side surface 102s by the magnetic force of the magnet 116 disposed close to the side surface 102s.

  The reaction liquid excluding the magnetic particles 311 is transferred to the recovery chamber 103 via the B channel 112. A certain amount of cleaning liquid is transferred from the A channel 111 to the B / F separation chamber 102, and the captured magnetic particles 311 are washed in the B / F separation chamber 102. The cleaning liquid is transferred to the collection chamber 103 via the B channel 112. As will be described in detail below, the space (second space) of the B / F separation chamber 102 includes a first region 102a and a second region 102b. The second region 102b is formed by B / F by capillary action. The reaction solution or the cleaning solution held in the separation chamber 102 can be sucked and held in the second region 102b. The side surface 102s is located in the second region 102b, and the B channel 112 is connected to the side surface 102s.

  The recovery chamber 103 stores a reaction liquid other than the magnetic particles 311 transferred from the B / F separation chamber 102 via the B channel 112 and a used cleaning liquid. In order to prevent these liquids from returning to the B / F separation chamber 102 more reliably, in the present embodiment, the recovery chamber 103 includes the first sub chamber 103A, the second sub chamber 103B, the first sub chamber 103A, and the second sub chamber 103A. A connecting portion 115 for connecting the sub chamber 103B is included. The connection part 115 is a flow path in which the liquid can move depending on the weight. The first sub chamber 103 </ b> A and the second sub chamber 103 </ b> B are each constituted by another independent space, and are connected by a connecting portion 115. That is, the space (third space) of the recovery chamber 103 includes the space of the first sub chamber 103A and the space of the second sub chamber 103B. In the present embodiment, the second sub chamber 103B is located farther from the rotation shaft 110 than the first sub chamber 103A. Further, the first sub chamber 103 </ b> A is located farther from the rotation shaft 110 than the B / F separation chamber 102. That is, the collection chamber 103 is located farther from the rotation shaft 110 than the B / F separation chamber 102 as a whole. The space of the first sub chamber 103A has a larger capacity than the larger amount of the reaction liquid and the amount of the cleaning liquid for one time. The second sub chamber 103B has a capacity larger than the sum of the reaction liquid and the cleaning liquid for a plurality of times.

  In the present embodiment, the first sub chamber 103A and the second sub chamber 103B each have a space extending along the circumferential direction of the substrate 100 ′, and the space inside the first sub chamber 103A and the second sub chamber 103B. The liquid absorbing member 121 and the liquid absorbing member 122 are respectively disposed. The sample analysis substrate 100 may include only one of the liquid absorbing member 121 and the liquid absorbing member 122.

  The sample analysis substrate 100 includes the liquid absorption member 121 and the liquid absorption member 122, so that the reaction liquid or the cleaning liquid transferred to the recovery chamber 103 flows back to the B / F separation chamber 102 via the B channel 112. Suppress. That is, the liquid transferred to the recovery chamber 103 is more reliably held inside.

  The liquid absorbing member 121 and the liquid absorbing member 122 are made of a material that absorbs a liquid such as a reaction liquid and a used cleaning liquid. For example, an absorbent polymer typified by sodium polyacrylate, a synthetic resin such as polyvinyl alcohol and urethane, or a sponge made of a natural material such as sponge, water absorbent paper such as filter paper, and the like are used as the liquid absorbent member 121 and the liquid absorbent member 122. Can be used. Here, absorption means that the liquid absorbing member can hold the liquid without moving the liquid due to the force even if a force of at least about gravity is applied.

  The liquid absorbing member 121 and the liquid absorbing member 122 are disposed at one end of a space extending along the circumferential direction of the first sub chamber 103A and the second sub chamber 103B. Preferably, the liquid absorbing member 121 is a space having a predetermined volume in the space of the first sub chamber 103A between the liquid absorbing member 121 and the connection portion between the B flow path 112 and the first sub chamber 103A. 103Aa. That is, the space 103 </ b> Aa in which the liquid absorbing member 121 is not provided is provided so that the opening of the B channel 112 connected to the first sub chamber 103 </ b> A is not blocked by the liquid absorbing member 121.

  Similarly, in the space of the second sub chamber 103B, the liquid absorbing member 122 is a space 103Ba having a predetermined volume between the liquid absorbing member 122 and a connection portion between the connecting portion 115 and the second sub chamber 103B. It is arranged to have. That is, the space 103Ba in which the liquid absorbing member 122 is not provided is provided so that the opening of the coupling portion 115 connected to the second sub chamber 103B is not blocked by the liquid absorbing member 122.

  Preferably, in the space of the first sub chamber 103A, the liquid absorbing member 121 is a space 103Ab having a predetermined volume between the liquid absorbing member 121 and a connection portion between the connecting portion 115 and the first sub chamber 103A. It is also arranged to have. That is, the space 103Ab in which the liquid absorbing member 121 is not provided is provided so that the opening of the connecting portion 115 connected to the first sub chamber 103A is not blocked by the liquid absorbing member 121.

  By providing the spaces 103Aa, 103Ab, and 103Ba, the reaction liquid and the cleaning liquid transferred from the B channel 112 can flow into the first sub chamber 103A without being blocked by the liquid absorbing member 121 that has absorbed the liquid. Further, the reaction liquid and the cleaning liquid in the first sub chamber 103A can flow into the second sub chamber 103B without being blocked by the liquid absorbing member 121 that has absorbed liquid and the liquid absorbing member 122 that has absorbed liquid.

  The liquid absorption member 121 and the liquid absorption member 122 are prevented from moving in the space of the first sub chamber 103A and the second sub chamber 103B depending on the rotation of the sample analysis substrate 100 or the angular position of the sample analysis substrate 100. It is preferable that the liquid absorbing member 121 and the liquid absorbing member 122 are fixed, or the movement of the liquid absorbing member 121 and the liquid absorbing member 122 is restricted so as not to enter the above-described spaces 103Aa, 103Ab, and 103Ba. For example, the liquid absorbing member 121 and the liquid absorbing member 122 are an adhesive or the like at one end of a space extending along the circumferential direction of the first sub chamber 103A and the second sub chamber 103B. It may be fixed by. Further, around the spaces 103Aa, 103Ab, and 103Ba, mesh-like partitions or walls that prevent the liquid absorbing member 121 and the liquid absorbing member 122 from entering and allow the liquid to move freely may be provided. .

  The size (volume) of the liquid absorbing member 121 and the liquid absorbing member 122 is not particularly limited, and is accommodated in a state where the above-described spaces 103Aa, 103Ab, and 103Ba are secured in the spaces of the first sub chamber 103A and the second sub chamber 103B. It only has to be done. As long as at least one of the liquid absorbing member 121 and the liquid absorbing member 122 is disposed, at least a part of the reaction liquid or the cleaning liquid transferred to the recovery chamber 103 is absorbed by the liquid absorbing member 121 or the liquid absorbing member 122. It is possible to suppress the liquid transferred to the recovery chamber 103 from flowing back to the B / F separation chamber 102 via the B channel 112 or the B channel 112 to the extent corresponding to the absorption amount. Further, if the absorbing member 121 and / or the liquid absorbing member 122 have a size capable of absorbing the entire amount of the reaction liquid and the cleaning liquid transferred to the recovery chamber 103, the above-described B channel 112 or B / F The back flow to the separation chamber 102 can be more reliably suppressed. Therefore, the sizes of the liquid absorbing member 121 and the liquid absorbing member 122 are based on specifications and designs such as the amount of reaction liquid and cleaning liquid transferred to the recovery chamber 103 and how much backflow should be suppressed. Can be arbitrarily determined.

  Next, each flow path will be described. The A channel 111 transfers the cleaning liquid stored in the cleaning liquid chamber 101 to the B / F separation chamber 102. At this time, instead of the total amount of cleaning liquid in the cleaning liquid chamber 101, the cleaning liquid is weighed once based on the capacity of the space defined by the A channel 111, and the weighed cleaning liquid is transferred to the B / F separation chamber 102. The A channel 111 includes a first opening 11c and a second opening 111d, the first opening 111c is connected to the cleaning liquid chamber 101, and the second opening 111d is connected to the B / F separation chamber 102. More specifically, the A channel 111 includes a first portion 111a having a first opening 111c and a second portion 111b having a second opening 111d. The first portion 111a and the second portion 111b are respectively connected at one end where the first opening 111c and the second opening 111d are not located.

  The first opening 111c is located closer to the rotation shaft 110 than the second opening 111d. Each part of the A channel 111 is preferably located at the same position as the first opening 111c from the rotation shaft 110 or farther from the rotation shaft 110 than the first opening 111c. Thus, when a centrifugal force acts on the cleaning liquid in a state where the A channel 111 is filled with the cleaning liquid, all the cleaning liquid in the A channel 111 is transferred to the B / F separation chamber 102 without returning to the cleaning liquid chamber 101. The

  The total capacity of the first portion 111a and the second portion 111b corresponds to the amount of the cleaning liquid for one time, and the space between the first opening 111c and the second opening 111d of the A channel 111 is seen by the cleaning liquid. Thus, the washing liquid for one time is weighed. Both the first portion 111a and the second portion 111b of the A channel 111 can be filled with the cleaning held in the cleaning liquid chamber 101 by capillary action. 3C shows a first portion 111a and a second portion 111b of the A channel 111 from the first region 101a of the cleaning liquid chamber 101, the first region 102a of the B / F separation chamber 102, and The thickness (depth) in the direction parallel to the thickness of the substrate 100 ′ of the space in the path connected to the B flow path 112 via the second region 102b is shown. In FIG. 3C, the horizontal axis indicates the distance from one end of the first region 101a, and the vertical axis indicates the thickness. In FIG. 3C, the horizontal axis is an example for explanation, and is not shown accurately. Similarly, the vertical axis shows the relative thickness relationship between adjacent regions, but the thickness values are not accurately shown.

  As shown in FIG. 3C, in the present embodiment, in the A channel 111, the thickness of the second portion 111b is smaller than the thickness of the first portion 111a, and the second portion 111b has a greater capillary force than the first portion 111a. Prepare. For this reason, when the cleaning liquid held in the first region 101a of the cleaning liquid chamber is sucked into the first portion 111a of the A channel 111 from the first opening 111c, the cleaning liquid reaches the second portion 111b on which a larger capillary force works. Go around. As a result, the entire A channel 111 is filled with the cleaning liquid.

  In the first region 101a of the cleaning liquid chamber 101, in FIG. 3C, the thickness of the connection portion 101c is smaller than the thickness of the other portion of the first region 101a. Further, the thickness of the connecting portion 101c is the same as the thickness of the first portion 111a. Thereby, it is possible to make capillary force act on the connection part 101c. As shown in FIG. 3B, the connecting portion 101c has an opening 101d (shown by a thick line) larger than the first opening 111c, and contacts the remaining portion of the first region 101a through the opening 101d. For this reason, even when the sample analysis substrate 100 is stopped at various rotation angles, a part of the opening 101d of the connection part 101c comes into contact with the cleaning liquid in the cleaning liquid chamber 101, so that the connection part 101c sucks the cleaning liquid, The connecting portion 101c can be filled with the cleaning liquid.

  In addition, the thickness of the connection part 101c may differ from the 1st part 111a. Further, the entire first region 101a of the cleaning liquid chamber 101 may be the connection part 101c or may not have the connection part 101c.

  The capillary force of the first portion 111a of the A channel 111 is larger than the capillary force of the connecting portion 101c. Therefore, the cleaning liquid held in the connecting portion 101c can be sucked and moved by the first portion 111a of the A channel 111. Further, since the opening 101d of the connecting portion 101c is larger than the first opening 111c of the A channel 111, the connecting portion 101c functions as a funnel, and a large amount of cleaning liquid smoothly passes through the connecting portion 101c. Can be aspirated.

  The B channel 112 can also suck the liquid held in the B / F separation chamber 102 by capillary action. As shown in FIG. 3C, the thickness of the B channel 112 is smaller than the thickness of the second region 102 b of the B / F separation chamber 102. In the B / F separation chamber 102, the second region 102 b has a thickness smaller than that of the first region 102 a and larger than that of the B channel 112. Therefore, it is possible to apply a capillary force to the second region 102b, and the cleaning liquid transferred from the A channel 111 is sucked into the second region 102b of the B / F separation chamber 102 by a capillary phenomenon. Since the B flow path 112 is connected to the second region 102b of the B / F separation chamber 102, a part of the cleaning liquid becomes B / F due to a capillary force larger than that of the second region 102b of the B / F separation chamber 102. Suction is performed from the F separation chamber 102 to the B flow path 112.

  The C channel 113 and the D channel 114 can also be filled with liquid by capillary action. Specifically, the C channel 113 and the D channel 114 can each be filled with a liquid filled in the storage chamber 104 and the reaction chamber 105 by capillary action.

  The B channel 112 and the D channel 114 can further control the movement of the liquid by the siphon principle. For this purpose, the B channel 112 has a first bent portion 112a and a second bent portion 112b. The first bent portion 112a has a convex shape on the side opposite to the rotary shaft 110, and the second bent portion 112b has a convex shape on the rotary shaft 110 side. The first bent portion 112a is between the B / F separation chamber 102 located on the side closer to the rotation shaft 110 in the B / F separation chamber 102 and the recovery chamber 103 to which the flow path is connected and the second bent portion 112b. Is located. Similarly, the D channel 114 has a first bent portion 114a and a second bent portion 114b. The first bent portion 114a has a convex shape on the side opposite to the rotary shaft 110, and the second bent portion 114b has a convex shape on the rotary shaft 110 side. The first bent portion 114a is located between the reaction chamber 105 and the B / F separation chamber 102 to which the flow path is connected, between the reaction chamber 105 located on the side closer to the rotation shaft 110 and the second bent portion 114b. ing.

  The distance between the rotation axis 110 and the side surface closest to the rotation axis of the chamber located far from the rotation axis 110 is R1, and from the rotation axis 110 to the point located farthest from the rotation axis 110 of the first bent portion When R2 is R2, it is preferable that R1> R2 (condition 1) is satisfied.

  In addition, when the liquid held in the rotation shaft 110 and the chamber located near the rotation shaft 110 is held biased to the side surface by centrifugal force, the distance from the rotation shaft to the liquid surface of the liquid is expressed as R4. When the distance from the rotating shaft 110 to the point located closest to the rotating shaft 110 of the second bent portion is R3, it is preferable that R4> R3 (condition 2) is satisfied.

As shown in FIG. 3D, conditions 1 and 2 are expressed as follows when distances R1 to R4 are set to distances 2R1 to 2R4 and 4R1 to 4R4 for the B channel 112 and the D channel 114, respectively.
B channel 112
Condition 1: 2R1> 2R2
Condition 2: 2R4> 2R3
D channel 114
Condition 1: 4R1> 4R2
Condition 2: 4R4> 4R3

  When the B channel 112 satisfies the conditions 1 and 2, when the reaction solution is transferred from the reaction chamber 105 to the B / F separation chamber 102 by centrifugal force, or from the A channel 111 to the B / F separation chamber When the cleaning liquid is transferred to 102, the reaction liquid or the cleaning liquid transferred to the B / F separation chamber 102 can be prevented from being transferred to the recovery chamber 103 as it is.

  Further, when the D channel 114 satisfies the conditions 1 and 2, the composite 310 can be formed in the reaction chamber 105, and the reaction liquid can be prevented from being transferred to the B / F separation chamber 102.

  When utilizing the capillary phenomenon, each flow path or chamber has a thickness of 50 μm to 300 μm, for example. In the case of forming chamber regions and flow paths having different thicknesses, for example, different thicknesses can be realized by changing the depths of the spaces provided in the base substrate 100a. Alternatively, the depth of the space provided in the base substrate 100a is made constant, and convex portions having different heights are provided at positions corresponding to the respective chambers and flow paths of the cover substrate 100b, so that the thicknesses of the respective flow paths and chambers are different. It may be allowed.

  Each of the cleaning liquid chamber 101, the B / F separation chamber 102, the recovery chamber 103, the storage chamber 104, and the reaction chamber 105 is provided with at least one air hole 108. Thereby, the inside of each chamber is maintained at atmospheric pressure, and each flow path can be moved by the capillary phenomenon and siphon principle. In addition, the reaction chamber 105 and the storage chamber 104 may be provided with an opening 109 for injecting a liquid such as a sample solution, a reaction solution, or a cleaning solution. The air hole 108 may also serve as the opening 109.

(Operation of sample analysis system 501)
The operation of the sample analysis system 501 will be described. FIG. 4 is a flowchart showing the operation of the sample analysis system 501. A program that defines the procedure for controlling each part of the sample analysis system 501 for operating the sample analysis system 501 is stored in, for example, the memory of the controller 205, and the following operations are performed by executing the program by the arithmetic unit. Realize. Prior to the following steps, the sample analysis substrate 100 is loaded into the sample analysis apparatus 200, and the origin of the sample analysis substrate 100 is detected.

[Step S1]
First, as shown in FIG. 5, the cleaning liquid is introduced into the storage chamber 104 of the sample analysis substrate 100. In addition, the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308 are introduced into the reaction chamber 105. For example, a liquid containing the magnetic particle-immobilized antibody 305 is held in the reaction chamber 105, and a chamber (not shown) provided on the sample analysis substrate 100 holds the liquid containing the antigen 306 and the labeled antibody 308 separately. These may be transferred to the reaction chamber 105 by a centrifugal force generated by the rotation of the sample analysis substrate 100. In the reaction chamber 105, the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308 are simultaneously reacted by an antigen-antibody reaction to form a complex 310. At this time, the C channel 113 and the D channel 114 are filled with the reaction solution containing the cleaning solution and the complex 310, respectively, by capillary action.

[Step S2]
After the complex 310 is generated, the sample analysis substrate 100 is rotated, and the reaction solution containing the complex 310 is moved to the B / F separation chamber 102. At this time, the D channel 114 is filled with the reaction solution by capillary action. For this reason, when the centrifugal force by rotation acts on the reaction liquid containing the complex 310 of the reaction chamber 105, the reaction liquid is transferred to the B / F separation chamber 102. The reaction liquid transferred to the B / F separation chamber 102 is not subsequently transferred to the recovery chamber 103 while the sample analysis substrate 100 is rotating. This is because, as described above, the B channel 112 forms a siphon, so that the liquid does not move in the direction toward the rotating shaft 110 against the centrifugal force against the centrifugal force. Of the reaction liquid containing the composite 310 transferred to the B / F separation chamber 102, most of the magnetic particles 311 are captured on the side surface 102 s by the magnetic force of the magnet 116.

  The rotation speed of the sample analysis substrate 100 is set such that a liquid such as a reaction liquid does not move due to gravity due to the centrifugal force generated by the rotation. Hereinafter, this rotation speed is set for rotation using centrifugal force.

  Simultaneously with the movement of the reaction liquid, the cleaning liquid is transferred from the storage chamber 104 to the cleaning liquid chamber 101 through the C channel 113.

  After the reaction solution and the cleaning solution are all transferred to the B / F separation chamber 102 and the cleaning solution chamber 101, the sample analysis substrate 100 is stopped at a predetermined angle. As shown in FIG. 6, the predetermined first angle means that the cleaning liquid transferred to the cleaning liquid chamber 101 in the sample analysis substrate 100 does not come into contact with the connection portion 101 c of the cleaning liquid chamber 101, and B / F This is the angle at which the reaction solution transferred to the separation chamber 102 can come into contact with the opening of the B channel 112. This angle depends on the shape of the cleaning liquid chamber 101 and the B / F separation chamber 102, the position in the substrate 100 ', the amounts of the cleaning liquid and the reaction liquid, the inclination angle θ of the sample analysis substrate 100, and the like. For example, in the example shown in FIG. 6, if the vertical direction (indicated by an arrow) in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 is within the angle range indicated by δ1 of the sample analysis substrate 100. Good.

  The reaction liquid in the B / F separation chamber 102 fills the B channel 112 by capillary action by contacting the opening of the B channel 112.

[Step S3 (Step (a))]
The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation, and acts on the reaction solution and the magnetic particles 311 (complex 310 and unreacted magnetic particle-immobilized antibody 305) in the B / F separation chamber 102. This centrifugal force acts so that the liquid and the complex move toward the side surface 102s of the B / F separation chamber 102. For this reason, as shown in FIG. 7, the magnetic particles 311 are pressed against the side surface 102s.

  The reaction solution that has received the centrifugal force is discharged from the B flow path 112 and transferred to the first sub chamber 103 </ b> A of the recovery chamber 103. At this time, a part of the reaction liquid may come into contact with the liquid absorbing member 121 located in the first sub chamber 103 </ b> A and be absorbed by the liquid absorbing member 121. The reaction solution is further transferred to the second sub chamber 103B through the connecting portion 115 from the opening of the connecting portion 115 connected to the first sub chamber 103A by centrifugal force. Part or all of the reaction liquid transferred to the second sub chamber 103B comes into contact with the liquid absorbing member 122 and is absorbed.

  Due to the sum of the centrifugal force and the attractive force of the magnet 116, the magnetic particles 311 are strongly pressed against the side surface 102s and captured. As a result, only the reaction solution is discharged from the B channel 112, and the magnetic particles 311 remain in the B / F separation chamber 102. The cleaning liquid in the cleaning liquid chamber 101 receives a centrifugal force due to rotation and moves to the second region 101b. After the transfer of the reaction liquid to the second sub chamber 103B is completed, the rotation of the sample analysis substrate 100 is stopped.

  Thereby, the reaction solution and the magnetic particles 311 are separated. Specifically, the reaction liquid moves to the second sub chamber 103 </ b> B of the recovery chamber 103, and the magnetic particles 311 remain in the B / F separation chamber 102. Even when the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the side surface 102s by the attractive force received from the magnet 116. The stop angle at this time may be the first angle, the second angle of the next step, or another angle.

[Step S4]
As shown in FIG. 8, the sample analysis substrate 100 is slightly rotated and stopped at a predetermined second angle. The second angle is an angle at which the cleaning liquid transferred to the cleaning liquid chamber 101 comes into contact with the connection portion 101 c of the cleaning liquid chamber 101. For example, in the example illustrated in FIG. 8, the vertical direction is an angle within the angle range indicated by δ <b> 2 of the sample analysis substrate 100.

  The cleaning liquid is sucked from the cleaning liquid chamber 101 by the capillary force in the connecting portion 101c, the first part 111a and the second part 111b of the A channel 111, and the first part 111a and the second part 111b of the A channel 111 are filled with the cleaning liquid. . Thereby, the washing | cleaning liquid for 1 time is weighed.

  In order to ensure that the A channel 111 is filled with the cleaning liquid, it may be rotated by several degrees alternately, that is, swung around the second angle clockwise and counterclockwise. Since capillary force acts on the A channel 111, the cleaning liquid does not move from the second portion 111 b of the A channel 111 to the B / F separation chamber 102 at this time.

  When the cleaning liquid moved to the second sub chamber 103B comes into contact with the liquid absorbing member 122 by stopping the sample analysis substrate 100 at the second angle, the cleaning liquid may be further absorbed by the liquid absorbing member 122.

[Step S5]
Subsequently, the sample analysis substrate 100 is rotated. The centrifugal force due to the rotation acts on the cleaning liquid in the A channel 111 and the cleaning liquid chamber 101. As shown in FIG. 9, the cleaning liquid in the A channel 111 is transferred to the B / F separation chamber 102 by centrifugal force. On the other hand, the excess cleaning liquid located in the first area 101a in the cleaning liquid chamber 101 moves to the second area 101b in the cleaning liquid chamber 101 by centrifugal force. Therefore, only the cleaning liquid weighed by the A channel 111 is transferred to the B / F separation chamber 102. Since the centrifugal force also acts on the cleaning liquid transferred to the B / F separation chamber 102, the cleaning liquid does not move in the direction of the rotation axis 110 in the B channel 112, and the cleaning liquid substantially remains in the B / F separation chamber 102. Thereby, the magnetic particles 311 in the B / F separation chamber 102 come into contact with the cleaning liquid, and the first cleaning is performed.

  As shown in FIG. 10, after all the cleaning liquid in the A channel 111 has moved to the B / F separation chamber 102, the sample analysis substrate 100 is stopped at a predetermined third angle. The third angle is an angle at which the cleaning liquid in the cleaning liquid chamber 101 does not come into contact with the connection part 101 c and the cleaning liquid transferred to the B / F separation chamber 102 can come into contact with the opening of the B flow path 112. . For example, in the example shown in FIG. 10, the vertical direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by δ3 on the sample analysis substrate 100.

  The cleaning liquid in the B / F separation chamber 102 contacts the opening of the B channel 112 and fills the B channel 112 by capillary action.

[Step S6]
The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation, and acts on the cleaning liquid and the magnetic particles 311 in the B / F separation chamber 102. This centrifugal force acts so that the cleaning liquid and the magnetic particles 311 move to the side surface 102 s of the B / F separation chamber 102, and the magnetic particles 311 are captured on the side surface 102 s by the centrifugal force and the attractive force by the magnet 116.

  The cleaning liquid that has received the centrifugal force is discharged from the B flow path 112 and transferred to the first sub chamber 103 </ b> A of the recovery chamber 103. At this time, a part of the cleaning liquid may come into contact with the liquid absorbing member 121 located in the first sub chamber 103 </ b> A and be absorbed by the liquid absorbing member 121. Further, the cleaning liquid is transferred to the second sub chamber 103B through the connecting portion 115. When the capacity of the liquid absorbing member 122 is large and a space capable of absorbing the liquid remains, a part or all of the transferred cleaning liquid is further absorbed by the liquid absorbing member 122.

  For this reason, as shown in FIG. 11, only the cleaning liquid is discharged from the B channel 112, and the magnetic particles 311 remain in the B / F separation chamber 102. The cleaning liquid in the cleaning liquid chamber 101 receives a centrifugal force due to rotation and moves to the second region 101b. After the transfer of the cleaning liquid to the second sub chamber 103B is completed, the rotation of the sample analysis substrate 100 is stopped. As a result, the cleaning liquid and the magnetic particles 311 are separated. Specifically, the cleaning liquid moves to the second sub chamber 103 </ b> B of the recovery chamber 103, and the magnetic particles 311 remain in the B / F separation chamber 102. Even when the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the side surface 102s by the attractive force received from the magnet 116. The stop angle at this time may be the third angle or the fourth angle of the next step.

[Step S7 (Process (e))]
As shown in FIG. 12, the sample analysis substrate 100 is slightly rotated and stopped at a predetermined fourth angle. The fourth angle is an angle at which the cleaning liquid transferred to the cleaning liquid chamber 101 comes into contact with the connection portion 101 c of the cleaning liquid chamber 101. For example, in the example shown in FIG. 12, the vertical direction is an angle within the angle range indicated by δ4 of the sample analysis substrate 100. Since the amount of the cleaning liquid remaining in the cleaning liquid chamber 101 in step S4 is different, the angle range δ4 may be different from the angle range δ2.

  The cleaning liquid is sucked from the cleaning liquid chamber 101 by the capillary force in the connecting portion 101c, the first part 111a and the second part 111b of the A channel 111, and the first part 111a and the second part 111b of the A channel 111 are filled with the cleaning liquid. . Thereby, the cleaning liquid for one time is weighed again.

  The sample analysis substrate 100 may be swung around the second angle so that the A channel 111 is reliably filled with the cleaning liquid. Since capillary force acts on the A channel 111, the cleaning liquid does not move from the second portion 111 b of the A channel 111 to the B / F separation chamber 102 at this time.

[Step S8]
Subsequently, the sample analysis substrate 100 is rotated. The centrifugal force due to the rotation acts on the cleaning liquid in the A channel 111 and the cleaning liquid chamber 101. As shown in FIG. 13, the cleaning liquid in the A channel 111 is transferred to the B / F separation chamber 102 by centrifugal force. On the other hand, the excess cleaning liquid located in the first area 101a in the cleaning liquid chamber 101 moves to the second area 101b in the cleaning liquid chamber 101 by centrifugal force. Therefore, only the cleaning liquid weighed by the A channel 111 is transferred to the B / F separation chamber 102. Since the centrifugal force also acts on the cleaning liquid transferred to the B / F separation chamber 102, the cleaning liquid does not move in the direction of the rotation axis 110 in the B channel 112, and the cleaning liquid substantially remains in the B / F separation chamber 102. Thereby, the magnetic particles 311 in the B / F separation chamber 102 come into contact with the cleaning liquid, and the second cleaning is performed.

  After all the cleaning liquid in the A channel 111 has moved to the B / F separation chamber 102, the sample analysis substrate 100 is stopped at a predetermined fifth angle as shown in FIG. The fifth angle is an angle at which the cleaning liquid in the cleaning liquid chamber 101 does not come into contact with the connection portion 101 c and the cleaning liquid transferred to the B / F separation chamber 102 can come into contact with the opening of the B flow path 112. . For example, in the example illustrated in FIG. 14, the vertical direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by δ5 on the sample analysis substrate 100.

  The cleaning liquid in the B / F separation chamber 102 contacts the opening of the B channel 112 and fills the B channel 112 by capillary action.

[Step S9]
The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation, and acts on the cleaning liquid and the magnetic particles 311 in the B / F separation chamber 102. This centrifugal force acts so that the cleaning liquid and the magnetic particles 311 move to the side surface 102 s of the B / F separation chamber 102, and the magnetic particles 311 are captured on the side surface 102 s by the centrifugal force and the attractive force by the magnet 116.

  The cleaning liquid that has received the centrifugal force is discharged from the B flow path 112 and transferred to the first sub chamber 103 </ b> A of the recovery chamber 103. Further, the cleaning liquid is transferred to the second sub chamber 103B through the connecting portion 115.

  For this reason, as shown in FIG. 15, only the cleaning liquid is discharged from the B channel 112, and the magnetic particles 311 remain in the B / F separation chamber 102. The cleaning liquid in the cleaning liquid chamber 101 receives a centrifugal force due to rotation and moves to the second region 101b. After the transfer of the cleaning liquid to the second sub chamber 103B is completed, the rotation of the sample analysis substrate 100 is stopped. As a result, the cleaning liquid and the magnetic particles 311 are separated. Specifically, the cleaning liquid moves to the second sub chamber 103 </ b> B of the recovery chamber 103, and the magnetic particles 311 remain in the B / F separation chamber 102. Even when the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the side surface 102s by the attractive force received from the magnet 116.

  At this time, a part of the cleaning liquid may come into contact with the liquid absorbing member 121 located in the first sub chamber 103 </ b> A and be absorbed by the liquid absorbing member 121. Further, in the second sub chamber 103 </ b> B, a part or all of the transferred cleaning liquid may be further absorbed by the liquid absorbing member 122.

  Through the above steps, B / F separation, specifically, the magnetic particles 311 and various unreacted substances and cleaning liquid are separated.

  Thereafter, a signal such as a dye, luminescence, or fluorescence corresponding to the labeling substance 307 of the labeled antibody 308 bound to the complex 310 included in the magnetic particle 311 is detected using the optical measuring device 207. Thereby, detection of the antigen 306, quantification of the concentration of the antigen 306, and the like can be performed.

(Liquid absorbing member)
The functions of the liquid absorbing members 121 and 122 in the movement of the reaction solution and the cleaning solution in the sample analysis substrate 100 described above will be described. FIG. 16A shows a state in which at least one of the reaction liquid and the cleaning liquid is transferred to the second sub chamber 103B in the sample analysis substrate that does not include the liquid absorbing members 121 and 122. The reaction liquid and the cleaning liquid are first transferred to the first sub chamber 103A, and then transferred to the second sub chamber 103B. At this time, a part of the reaction liquid or the cleaning liquid may remain in the first sub chamber 103A for some reason. Further, since the liquid can move in the connecting portion 115 by gravity, depending on the stop angle position of the sample analysis substrate 100, the reaction liquid or the cleaning liquid in the second sub chamber 103B may be transferred via the connecting portion 115. There is a case where the gas flows backward to the chamber 103A.

  In this case, as shown in FIG. 16B, when the sample analysis substrate 100 is stopped so that the connection portion between the B channel 112 and the first sub chamber 103A is positioned below the vertical direction, the inside of the first sub chamber 103A is stopped. The reaction solution and the cleaning solution fill the B channel 112 by capillary force. For this reason, there is a possibility that the reaction liquid or the cleaning liquid in the B / F separation chamber 102 cannot be transferred to the first sub chamber 103 </ b> A of the recovery chamber 103.

  On the other hand, as shown in FIG. 3B, when the first sub chamber 103A includes the liquid absorbing member 121, at least a part of the reaction liquid or the cleaning liquid remaining in the first sub chamber 103A is removed from the liquid absorbing member 121. Absorbs. For this reason, it is possible to reduce the possibility that the reaction liquid or the cleaning liquid in the first sub chamber 103 </ b> A flows backward to the B channel 112.

  When the liquid absorption member 122 is provided in the second sub chamber 103B, the liquid absorption member 121 absorbs at least a part of the reaction liquid or the cleaning liquid in the second sub chamber 103B. For this reason, it is possible to reduce the possibility that the reaction liquid or the cleaning liquid in the second sub chamber 103B moves to the first sub chamber 103A through the connecting portion 115 and further flows back to the B flow path 112.

  That is, the liquid absorbing member 121 and the liquid absorbing member 122 hold the liquid transferred to the recovery chamber 103 more reliably inside. This function can be exhibited independently of other features of the sample analysis substrate 100 of the present embodiment, but is preferably used when the reaction solution, the cleaning solution, etc. are moved to the collection chamber 103 in two or more steps. It is done. When the reaction liquid or the cleaning liquid is transferred to the recovery chamber 103 a plurality of times, the sample analysis substrate 100 may stop at various angular positions after the reaction liquid or the first cleaning liquid has moved to the recovery chamber 103. This is because the reaction liquid or the cleaning liquid may flow back to the B / F separation chamber 102 as described above depending on the angle position.

  As described above, according to the sample analysis substrate, sample analysis apparatus, and sample analysis system of the present embodiment, the liquid can be introduced into the same chamber in a plurality of times. For this reason, when performing B / F separation using the sample analysis substrate, sufficient cleaning can be performed. When the liquid is weighed, a flow path that develops a capillary force is used, so that each weighing can be performed more reliably and accurately. This operation can be realized by controlling the rotation and stop of the sample analysis substrate and controlling the angle at the time of stop. For this reason, it can be suitably applied to analysis methods in which components in a sample are analyzed through complex reaction steps including B / F separation without using a large analytical instrument or manually operated by an operator. It is.

  In addition, the shape and arrangement of each chamber and flow path of the sample analysis substrate illustrated in the above embodiment are merely examples, and various modifications can be made. For example, the sample analysis substrate 150 shown in FIG. 17 has a cleaning liquid chamber 101 including a first region 101a 'and a second region 101b. Unlike the sample analysis substrate 100 shown in FIG. 3B, the first region 101a ′ does not have the connection portion 101c and extends from the first opening 111c in the first region toward the side farther from the rotation shaft 110. Contains only part. The entire first region 101a 'can suck the liquid held in the second region 101b by capillary action. The cleaning liquid held in the cleaning liquid chamber 101 can be sucked and held by the capillary action as a whole. One end of the first region 101a ′ is connected to the first opening 111c of the A channel 111, and the second region 101b is connected to the first region 101a at a position farther from the rotation shaft 110 than the first opening 111c. Yes. As described above, the sample analysis substrate 150 can weigh the cleaning liquid for one time by the A channel 111. According to the sample analysis substrate 150, if the portion of the first region 101a ′ connected to the second region 101b is in contact with the cleaning liquid, the first region 101a ′ and the A channel 111 are filled with the cleaning liquid by capillary force. Can do. For this reason, it becomes possible to fill the A channel 111 with the cleaning liquid in a rotation angle range wider than that of the sample analysis substrate 100, and the first angle range δ1 and the second angle range δ2 become wide. Therefore, the degree of freedom of control in the sample analysis system is increased.

  As shown in FIG. 18, the sample analysis substrate 160 may further include liquid absorbing members 121 'and 122' located in the first sub chamber 103A and the second sub chamber 13B, respectively.

  The liquid absorbing member 121 ′ is provided in the space in the first sub chamber 103A so as to leave the space 103Ab along the wall surface 103As located on the side far from the rotation axis. . Similarly, the liquid absorbing member 122 'is provided along the wall surface 103Bs located on the far side from the rotation axis in the space in the second sub chamber 103B.

  According to this configuration, the reaction liquid and a part of the cleaning liquid transferred to the first sub chamber 103A and the second sub chamber 13B are surely brought into contact with and absorbed by the liquid absorbing member 121 ′ and the shape absorbing member 122 ′. The Therefore, the possibility that the reaction liquid or the cleaning liquid in the first sub chamber 103 </ b> A and the second sub chamber 103 </ b> B flows backward to the B channel 112 can be further reduced.

  3B and FIG. 18, the number of liquid absorbing members arranged in the first sub chamber 103A and the second sub chamber 13B is one, but two or more liquid absorbing members are arranged in the first sub chamber 103A. In one or both of the second sub chamber 13B and the second sub chamber 13B, the second sub chamber 13B may be disposed at an adjacent position or a different position.

  In the present embodiment, the measurement system using magnetic particles has been described. However, the sample analysis substrate, the sample analysis apparatus, the sample analysis system, and the sample analysis system program according to one embodiment of the present application are magnetic. It is not limited to a measurement system using particles. For example, the target to which the primary antibody is immobilized may be a wall surface in the chamber instead of the magnetic particles. That is, when the chamber is made of a material such as polystyrene or polycarbonate, the primary antibody can be immobilized on the wall surface in the chamber by physical adsorption, and sandwiched with antigen or labeled antibody in the chamber. The reaction can be triggered. Moreover, if it is the structure which equips the wall surface in a chamber with a metal substrate, a primary antibody can be couple | bonded and fixed to a metal substrate using SAM, for example, and it is a sandwich type with an antigen and a labeled antibody in a chamber. It is possible to cause a binding reaction. When the primary antibody is immobilized on the wall surface of the chamber by physical adsorption, it is mainly used in a system for detecting a dye, chemiluminescent or fluorescent signal. On the other hand, when the primary antibody is immobilized on a metal substrate, it is mainly used as a signal in a system for detecting an electrochemical signal (for example, current) or an electrochemiluminescence signal. In this case, the magnet 116 shown in FIG. 3B is unnecessary. The reaction field for forming the composite 310 is not the reaction chamber 105 but the B / F separation chamber 102. Therefore, the primary antibody needs to be immobilized on the wall surface of the B / F separation chamber 102.

  In the above embodiment, an example of cleaning for B / F separation has been described. However, the sample analysis substrate, the sample analysis apparatus, and the sample analysis system of this embodiment divide a solution other than the cleaning liquid into a plurality of times as described above. It is applicable to various sample analysis methods introduced into the same chamber. In the above embodiment, the liquid is continuously introduced into the chamber. However, by appropriately controlling the rotation and stop of the sample analysis substrate and the angle at the time of stop, the other can be performed in between. It is also possible to include a process.

  In the above embodiment, the cleaning is performed twice, but may be performed three or more times as necessary.

  In the above embodiment, the sample analysis substrate includes the liquid absorbing member positioned in the recovery chamber 103. However, as described above, the configuration including the liquid absorbing member and the A channel including the weighing function are provided. It can be implemented independently of the configuration and the like. For example, the sample analysis substrate may not include the liquid absorption member, and the sample analysis substrate may include the liquid absorption member, but may not include the configuration of the A channel having the weighing function. . In this case, the reaction liquid or the cleaning liquid can be moved from the B / F separation chamber 102 to the recovery chamber 103 only by centrifugal force without depending on the gravity, so that the liquid absorbing members 121 and 122 in the recovery chamber 103 can be moved. It is possible to hold a reaction solution or a washing solution. Therefore, the sample analysis substrate may be rotated around the rotation axis in a state where the rotation axis 110 is at an angle of 0 ° to 90 ° with respect to the direction of gravity.

  The sample analysis substrate, sample analysis apparatus, sample analysis system, and sample analysis system program disclosed in the present application can be applied to analysis of a specific component in a specimen using various reactions.

100, 150, 160 Sample analysis substrate 100 ′ Substrate 100a Base substrate 100b Cover substrate 101 Cleaning liquid chamber 101a First region 101a ′ First region 101b Second region 101c Connection portion 101d Opening 102 B / F separation chamber 102a First region 102b Second region 102s Side surface 103 Recovery chamber 103A First sub chamber 103B Second sub chamber 104 Storage chamber 105 Reaction chamber 108 Air hole 109 Opening 110 Rotating shaft 111 A Flow path 111a First portion 111b Second portion 111c First opening 111d First 2 opening 112 B flow path 112a 1st bending part 112b 2nd bending part 113 C flow path 114 D flow path 114a 1st bending part 114b 2nd bending part 115 Connection part 116 Magnet 121,122 Liquid absorption member 200 Sample analyzer 201 Motor 201a Turntable 203 Origin detector 204 Angle detector 205 Controller 206 Driver 207 Optical measuring device 302 Magnetic particle 304 Primary antibody 305 Magnetic particle immobilized antibody 306 Antigen 307 Labeled substance 308 Labeled antibody 310 Complex 311 Magnetic particle 501 Sample Analysis system

Claims (11)

A sample analysis substrate that transfers liquid by a rotational motion,
A substrate having a plate shape having a rotation axis and a predetermined thickness;
A first chamber located within the substrate and having a first space for holding a liquid;
A second chamber having a second space for holding the liquid discharged from the first chamber in the substrate;
A first flow path located in the substrate and having a path connecting the first chamber and the second chamber;
A liquid absorbing member disposed in the second space of the second chamber and capable of absorbing at least part of the liquid discharged into the second chamber;
A substrate for sample analysis comprising:   The liquid absorbing member has a third space having a predetermined volume between the liquid absorbing member and a connection portion between the first flow path and the second chamber in the second space. The sample analysis substrate according to claim 1, which is arranged.   The sample analysis substrate according to claim 1 or 2, wherein the first channel can fill the first channel with a liquid held in the first space by capillary action. The second chamber includes a first sub chamber, a second sub chamber, and a connecting portion that connects the first sub chamber and the second sub chamber,
4. The sample analysis substrate according to claim 1, wherein the second sub chamber is located farther from the rotation axis than the first sub chamber. 5.   The sample analysis substrate according to claim 4, wherein the liquid absorbing member is disposed in the first sub chamber.   The sample analysis substrate according to claim 4, wherein the liquid absorbing member is disposed in the second sub chamber. A third chamber located in the substrate and having a third space for holding a liquid;
A second flow path located in the substrate, having a path connecting the third chamber and the first chamber, and capable of being filled with a liquid held in the third space by capillary action; ,
With
The second flow path has a first opening and a second opening, and the first opening and the second opening are connected to the third chamber and the first chamber, respectively, and the first opening is the second opening. Located closer to the axis of rotation than
The first space includes a first region and a second region, and the first region includes a portion connected to the first opening and extending from the first opening toward a side farther from the rotation axis, The second region is connected to the extending portion of the first region at a position farther from the rotation axis than the first opening,
The sample analysis substrate according to claim 1, wherein the third space of the third chamber is larger than a volume of the second flow path.   The sample analysis substrate according to claim 1, further comprising a magnet positioned in proximity to the first chamber. The sample analysis substrate according to any one of claims 1 to 8,
A motor that rotates the sample analysis substrate around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity;
A rotation angle detector for detecting a rotation angle of the rotation shaft of the motor;
Based on the detection result of the rotation angle detector, a driver that controls the rotation angle of the motor when rotating and stopping, and an arithmetic unit, a memory, and a program stored in the memory and configured to be executable by the arithmetic unit A sample analyzer having a controller for controlling operations of the motor, the rotation angle detector, the origin detector, and the driver, based on the program,
A sample analysis system comprising:
The program is
When the sample analysis substrate in which the liquid in the first chamber is filled is loaded in the sample analysis device,
(A) By rotating the sample analysis substrate, at least a part of the liquid in the first chamber is transferred to the second chamber via the first channel;
Sample analysis system. A motor that rotates the sample analysis substrate according to any one of claims 1 to 8 around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity.
A rotation angle detector for detecting a rotation angle of the rotation shaft of the motor;
Based on the detection result of the rotation angle detector, a driver that controls the rotation angle of the motor when rotating and stopping, and an arithmetic unit, a memory, and a program stored in the memory and configured to be executable by the arithmetic unit A sample analyzer having a controller for controlling operations of the motor, the rotation angle detector, the origin detector, and the driver, based on the program,
With
The program is
When the sample analysis substrate in which the liquid in the first chamber is filled is loaded in the sample analysis device,
(A) A sample analyzer that transfers at least a part of the liquid in the first chamber to the second chamber via the first flow path by rotating the sample analysis substrate. The sample analysis substrate according to any one of claims 1 to 8,
A motor that rotates the sample analysis substrate around the rotation axis in a state where the rotation axis is at an angle of 0 ° to 90 ° with respect to the direction of gravity;
A rotation angle detector for detecting a rotation angle of the rotation shaft of the motor;
Based on the detection result of the rotation angle detector, a driver that controls the rotation angle of the motor when rotating and stopping, and an arithmetic unit, a memory, and a program stored in the memory and configured to be executable by the arithmetic unit A sample analyzer having a controller for controlling operations of the motor, the rotation angle detector, the origin detector, and the driver, based on the program,
A sample analysis system program comprising:
The program is
When the sample analysis substrate in which the liquid in the first chamber is filled is loaded in the sample analysis device,
(A) By rotating the sample analysis substrate, at least a part of the liquid in the first chamber is transferred to the second chamber via the first channel;
Sample analysis system program.

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