WO2023090446A1 - Analyseur d'échantillon et procédé d'analyse d'échantillon - Google Patents

Analyseur d'échantillon et procédé d'analyse d'échantillon Download PDF

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Publication number
WO2023090446A1
WO2023090446A1 PCT/JP2022/043035 JP2022043035W WO2023090446A1 WO 2023090446 A1 WO2023090446 A1 WO 2023090446A1 JP 2022043035 W JP2022043035 W JP 2022043035W WO 2023090446 A1 WO2023090446 A1 WO 2023090446A1
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WIPO (PCT)
Prior art keywords
container
solid phase
reagent
sample analyzer
sample
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PCT/JP2022/043035
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English (en)
Japanese (ja)
Inventor
敏博 笠間
亮 三宅
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国立大学法人東京大学
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Application filed by 国立大学法人東京大学 filed Critical 国立大学法人東京大学
Priority to JP2023562431A priority Critical patent/JPWO2023090446A1/ja
Publication of WO2023090446A1 publication Critical patent/WO2023090446A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations

Definitions

  • the present disclosure relates to sample analyzers and sample analysis methods.
  • This sample analysis method is a method of analyzing a sample using a microdevice provided with at least one channel.
  • a microstructure holding a specific binding reagent is provided in the channel.
  • the channel is washed with a cleaning liquid.
  • the channel is washed with a washing liquid.
  • the components of the sample are analyzed based on whether or not the reagent containing the enzyme substrate develops color when the channel of the microdevice is filled.
  • An object of the present disclosure is to provide a sample analyzer and sample analysis method capable of improving the accuracy of analysis results.
  • a sample analyzer is a sample analyzer that analyzes a sample, and includes a solid phase, a container, and a drive unit.
  • a specific binding reagent that specifically binds to the target substance contained in the specimen is immobilized on the solid phase.
  • the container is provided with a plurality of storage units in which a specimen, a detection reagent used for detecting a target substance, and a washing liquid are individually placed.
  • the driving section moves at least one of the solid phase and the container to immerse and take out the solid phase from each of the plurality of storage sections.
  • a specimen detection method is a specimen analysis method for analyzing a specimen, comprising a step of immobilizing a specific binding reagent that specifically binds to a target substance contained in the specimen on a solid phase; a step of individually placing the sample, a detection reagent used for detecting a target substance contained in the sample, and a washing solution into a plurality of storage units provided in the plurality of storage units; and immersing and removing the solid phase from each of the receptacles of the.
  • the solid phase is immersed in and removed from each of the plurality of storage units by the driving unit, so manual work by the operator is almost unnecessary.
  • it is possible to eliminate the influence of differences in work skills of workers on analysis results as much as possible, so that it is possible to improve the accuracy of analysis results.
  • FIG. 1 is a perspective view showing the perspective structure of the sample analyzer of the first embodiment.
  • FIGS. 2A to 2E schematically show how the target substance, the detection antibody, and the enzyme-labeled antibody bind to the primary antibody immobilized on the solid phase, and how the enzyme substrate activates the enzyme. It is a diagram.
  • FIG. 3 is a plan view showing the planar structure of the container of the first embodiment.
  • 4 is a side view showing the side structure of the container viewed from arrow V1 in FIG. 3.
  • FIG. 5 is a side view showing the side structure of the container as seen from arrow V2 in FIG. 3.
  • FIG. FIG. 6 is a block diagram showing the electrical configuration of the sample analyzer of the first embodiment.
  • FIG. 3 is a plan view showing the planar structure of the container of the first embodiment.
  • 4 is a side view showing the side structure of the container viewed from arrow V1 in FIG. 3.
  • FIG. 5 is a side view showing the side structure of the container as seen from
  • FIGS. 7 is a flowchart illustrating the procedure of processing executed by a control unit according to the first embodiment
  • FIGS. 8A and 8B are plan views showing an operation example of the container of the first embodiment.
  • FIGS. 9A to 9C are side views showing an example of the operation of the solid phase of the first embodiment.
  • 10A and 10B are plan views showing an operation example of the container of the first embodiment.
  • FIGS. 11A to 11C are side views showing an operation example of the solid phase of the first embodiment.
  • 12(A) and (B) are plan views showing an operation example of the container of the first embodiment.
  • 13A and 13B are plan views showing an operation example of the container of the first embodiment.
  • FIG. 14 is a plan view showing the planar structure of the container of the second embodiment.
  • FIG. 15 is a front view showing the front structure of the tube of the second embodiment.
  • FIG. 16 is a block diagram showing the electrical configuration of the sample analyzer of the third embodiment.
  • FIG. 17 is a front view showing the front structure of the sample analyzer of the fourth embodiment.
  • FIG. 18 is a plan view showing the planar structure of the container of the fourth embodiment.
  • FIG. 19 is a front view showing the front structure of the sampling instrument of the fourth embodiment.
  • FIG. 20 is a cross-sectional view showing the cross-sectional structure of the container and rotating portion of the fourth embodiment.
  • FIG. 21 is a bottom view showing the bottom structure of the container of the fourth embodiment.
  • FIG. 22 is a plan view showing the planar structure of the rotating portion of the fourth embodiment.
  • FIG. 23 is a front view showing an operation example of the sample analyzer of the fourth embodiment.
  • FIG. 24 is a plan view showing a planar structure of a container of a modified example of the fourth embodiment.
  • FIG. 25 is an enlarged side view showing the side structure around the well of the container of the fifth embodiment.
  • FIG. 26 is a front view showing an enlarged structure around the tip of the solid phase of the fifth embodiment.
  • FIG. 27 is a cross-sectional view showing cross-sectional structures of a container and a solid phase of the fifth embodiment.
  • FIG. 28 is a front view showing the front structure of the sample analyzer of the sixth embodiment.
  • FIG. 29 is a plan view showing the planar structure of the container of the sixth embodiment.
  • FIG. 30 is a plan view showing the planar structure of the sample analyzer of the seventh embodiment.
  • FIG. 31 is a side view showing the side structure of the sample analyzer of the seventh embodiment.
  • FIG. 32 is a perspective view showing the perspective structure of the sample analyzer of the eighth embodiment.
  • FIG. 33 is a side view showing the side structure of the container of the eighth embodiment.
  • a sample analyzer 10 of the present embodiment shown in FIG. 1 is a device that analyzes components contained in a sample. Any specimen can be used as long as it may contain the test substance. For example, blood, saliva, nasopharyngeal swab, pleural effusion tissue section lysate from lung cancer patients, brain tumor lysate, urine, milk and the like can be used as specimens.
  • the sample analyzer 10 includes a solid phase 20 , a container 30 , a first driving device 40 and a second driving device 50 .
  • the solid phase 20 is made of acrylic resin or the like and formed in a cylindrical rod shape.
  • the outer diameter of the solid phase 20 of this embodiment is set to 1 [mm].
  • a specific binding reagent is immobilized on the solid phase 20 .
  • a specific binding reagent specifically binds to a target substance (molecule) such as a target protein contained in a specimen.
  • Specific binding reagents are antibodies, single-stranded nucleic acids, enzymes, and the like.
  • a primary antibody 101 contained in a specific binding reagent is immobilized on the surface of solid phase 20 .
  • the proximal end of the solid phase 20 is attached to the distal end of a cylindrical mounting member 21 .
  • a proximal end portion of the mounting member 21 is gripped and fixed by the second driving device 50 .
  • the container 30 contains a sample to be analyzed, a detection reagent used for detecting a target substance contained in the sample, a washing liquid, and the like.
  • a detection antibody reagent is a reagent containing an unlabeled detection antibody (secondary antibody) that binds to the target substance of the specimen.
  • the labeled antibody reagent is a reagent containing an enzyme-labeled antibody (tertiary antibody), such as an HRP (Horse Radish Peroxidase)-labeled antibody, that binds to the detection antibody.
  • a substrate reagent is a reagent that contains an enzyme substrate, such as the TMB (3,3',5,5'-Tetramethyl Benzidine) substrate.
  • an enzyme substrate such as the TMB (3,3',5,5'-Tetramethyl Benzidine) substrate.
  • the target substance 100 contained in the specimen binds to the primary antibody 101 as shown in FIG. 2(B).
  • the detection antibody 102 contained in the detection antibody reagent binds to the target substance 100 as shown in FIG. 2(C).
  • the enzyme-labeled antibody 103 contained in the labeled antibody reagent further binds to the detection antibody 102 as shown in FIG. 2(D). .
  • the substrate reagent is dissolved by the enzymatic activity of the enzyme-labeled antibody 103 against the substrate 104 contained in the substrate reagent, as shown in FIG. 2(E). develop color.
  • the more the target substance contained in the sample that is, the higher the concentration of the target substance in the sample, the more the detection antibody 102 and the enzyme-labeled antibody 103 that bind to the target substance. Therefore, the color development of the substrate reagent is enhanced, in other words, the absorbance is increased.
  • the washing liquid is used to wash the solid phase 20 removed from the specimen, the detection antibody reagent, and the labeled antibody reagent to wash off unreacted target substances, the detection antibody, and the labeled antibody from the solid phase 20.
  • buffers such as phosphate buffer, Tris buffer, carbonate buffer, PBS (Phosphate Buffered Saline), and TBS (Tris Buffered Saline) can be used.
  • the washing solution may contain a surfactant such as Tween or TritonX.
  • the container 30 is shaped like a disk around a predetermined axis m11.
  • First to eighth wells 31 to 38 are provided on the outer edge of the upper surface 300 of the container 30 .
  • the first to eighth wells 31 to 38 are arranged side by side in the circumferential direction A around the axis m11.
  • the first to eighth wells 31 to 38 are open to the upper surface 300 of the container 30 and are formed in a concave shape.
  • a specimen is placed in the first well 31 .
  • a third well 33 contains a detection antibody reagent.
  • a fifth well 35 contains a labeled antibody reagent.
  • a seventh well 37 contains a substrate reagent.
  • a second well 32, a fourth well 34, and a sixth well 36 contain a washing solution.
  • the eighth well 38 does not contain any reagent.
  • the upper surface 300 of the container 30 corresponds to the plane of the container 30 provided in the direction Z in which the predetermined axis m11 shown in FIG. 1 extends.
  • the wells 31 to 37 correspond to the accommodating portion. Specifically, the well 31 corresponds to the specimen containing portion, the wells 32, 34 and 36 correspond to the washing solution containing portion, and the wells 33, 35 and 37 correspond to the reagent containing portion. Of the wells 33, 35, and 37, the well 33 further corresponds to the detection antibody containing portion, the well 35 further corresponds to the labeled antibody containing portion, and the well 37 further corresponds to the substrate containing portion.
  • the direction Z in which the axis m11 extends is set to the vertical direction.
  • the vertical direction Z is hereinafter also referred to as the “vertical direction Z”.
  • the vertically upward direction Z1 is called “upward Z1”
  • the vertically downward direction Z2 is called “downward Z2”.
  • the well 31 is a concave groove extending in a semicircular shape around the axis m11.
  • the width of the well 31 in this embodiment is set to 2 [mm].
  • the wells 32 to 38 are arranged in order from the end 311 of the well 31 in the circumferential direction A at predetermined angular intervals.
  • Wells 32-38 are circular concave grooves.
  • the inner diameters of the wells 32 to 38 of this embodiment are set to 2 [mm].
  • FIG. 4 shows the side structure of the container 30 when viewed from the direction V1 shown in FIG. 5 shows the side structure of the container 30 when viewed from the direction V2 shown in FIG.
  • wells 31, 33, 35 and 37 have the same depth D1.
  • wells 32, 34, 36 and 38 have the same depth D2.
  • the depth D1 is set to 6 [mm] and the depth D2 is set to 11 [mm].
  • the volumes of the wells 33, 35 and 37 are set to 10 [ ⁇ L]
  • the volumes of the wells 32, 34 and 36 containing the washing liquid are set to 20 [ ⁇ L].
  • the volume of the well 31 into which the sample is placed is set to 1 [mL].
  • the depth D2 of the wells 32, 34, and 36 is deeper than the depth D1 of the wells 31, 33, 35, and 37.
  • a larger portion of the solid phase 20 is immersed in the washing liquid than in the case of immersing the solid phase 20 in the sample and detection reagent. Therefore, for example, by immersing the solid phase 20 in the well 31 and then immersing the solid phase 20 in the well 32, the specimen adhering to the solid phase 20 can be more reliably washed with the washing liquid.
  • a projecting portion 39 is formed on the outer peripheral surface of the container 30 .
  • Projection 39 is used to detect the rotational position of container 30 .
  • the projecting portion 39 corresponds to the marking portion.
  • the first drive device 40 includes a rotating portion 41 and a motor device 42 .
  • the first driving device 40 corresponds to the first driving section.
  • the rotating portion 41 is formed in a disk shape centering on the axis m11.
  • a container 30 is detachably attached to the upper surface of the rotating portion 41 .
  • the motor device 42 rotates the rotating portion 41 around the axis m11 based on the supply of electric power.
  • the motor device 42 is, for example, a stepping motor. As the rotating portion 41 rotates, the container 30 rotates together with the rotating portion 41 around the axis m11.
  • the second driving device 50 includes a pedestal 51 , a movable portion 52 and a motor device 53 .
  • the second driving device 50 corresponds to the second driving section.
  • the movable portion 52 is formed in a substantially rectangular parallelepiped shape.
  • a gripping portion 520 for gripping the proximal end portion of the mounting member 21 is formed on the front surface of the movable portion 52 .
  • a support member 510 , a rotating portion 511 and a linear rail 512 are provided on the upper surface of the pedestal 51 .
  • the support member 510 is provided to extend upward Z1 from the upper surface of the pedestal 51 .
  • the movable portion 52 is attached to the support member 510 so as to be slidable in the vertical direction Z. As shown in FIG.
  • the rotating part 511 is provided so as to protrude from the upper surface of the pedestal 51 .
  • the rotating portion 511 is formed in an annular shape around the axis m12.
  • a female screw portion is formed on the inner peripheral surface of the rotating portion 511 .
  • the rotating portion 511 rotates in the rotation direction B around the axis m12 based on the power transmitted from the motor device 53 .
  • the linear rail 512 is a rod-shaped member provided to extend upward Z1 from the inside of the rotating portion 511 .
  • An upper end portion of the linear rail 512 is fixed to the bottom surface of the movable portion 52 .
  • a male threaded portion 512 a is formed on the outer peripheral surface of the linear rail 512 .
  • the male threaded portion 512 a of the linear rail 512 is fitted to the female threaded portion of the inner peripheral surface of the rotating portion 511 .
  • the motor device 53 rotates the rotating portion 511 around the axis m12 based on the supply of electric power.
  • the motor device 53 is, for example, a stepping motor.
  • the rotating portion 511 rotates in the rotating direction B about the axis m12
  • the linear rail 512 fitted to the inner peripheral surface of the rotating portion 511 via the female threaded portion and the male threaded portion 512a moves straight in the vertical direction Z. physically displaced.
  • the movable portion 52 is displaced in the vertical direction Z together with the linear rail 512 .
  • the solid phase 20 is displaced in the vertical direction Z together with the movable portion 52 .
  • the sample analyzer 10 further includes a rotation sensor 60 and a control device 70.
  • the rotation sensor 60 is a sensor that detects the rotational position of the container 30 about the axis m11. As shown in FIG. 3 , the rotation sensor 60 is arranged so as to face the outer peripheral surface of the container 30 .
  • the rotation sensor 60 outputs an ON signal when the projecting portion 39 of the container 30 is positioned within its front detection range, and outputs an ON signal when the projecting portion 39 of the container 30 is not positioned within its front detection range. outputs an off signal. Therefore, based on whether the output signal of the rotation sensor 60 is an ON signal or an OFF signal, it can be determined whether the container 30 is positioned at the rotation position shown in FIG.
  • the rotational position of the container 30 shown in FIG. 3 is the position where the solid phase 20 faces one end 310 of the well 31 as shown in FIG. In this embodiment, the rotational position of the container 30 shown in FIGS. 1 and 3 is set as the initial position.
  • the control device 70 has a processor, a storage device, etc. as a hardware configuration.
  • the processor is a CPU (Central Processing Unit), a GPU (Graphical Processing Unit), or the like.
  • the storage devices are memories, HDDs (Hard Disk Drives) and/or SSDs (Solid State Drives).
  • the control device 70 has a position detection section 71 and a control section 72 as functional configurations realized by the processor executing a program stored in the storage device. .
  • the position detection unit 71 detects whether the container 30 is positioned at the initial position based on whether the output signal of the rotation sensor 60 is an ON signal or an OFF signal.
  • the control unit 72 controls the first driving device 40 and the second driving device 50 . Specifically, the control unit 72 drives the first driving device 40 to rotate the container 30 so that the wells 31 to 37 are sequentially positioned below the solid phase 20 . Further, the control unit 72 drives the second driving device 50 to displace the solid phase 20 in the vertical direction Z, thereby immersing and removing the solid phase 20 from the well positioned below the solid phase 20. conduct.
  • the control unit 72 places the wells 31 to 37 in order below the solid phase 20 and immerses and takes out the solid phase 20 from each of the wells 31 to 37 . As a result, the solid phase 20 is immersed in and taken out of each of the sample, washing solution, detection antibody reagent, washing solution, labeled antibody reagent, washing solution, and substrate reagent in order.
  • the controller 72 first drives the first driving device 40 to move the container 30 to the initial position shown in FIG. 8A (step S10). Subsequently, the controller 72 drives the second driving device 50 to immerse the solid phase 20 in the one end 310 of the first well 31 as shown in FIG. 9A (step S11 in FIG. 7). ). Subsequently, the control unit 72 drives the first driving device 40 to rotate the container 30 in the clockwise direction C shown in FIG. While maintaining this state, the solid phase 20 is moved to the other end 311 of the first well 31 as shown in FIGS.
  • step S12 in FIG. 7 the solid phase 20 comes into contact with the specimen placed in the first well 31 , and the target substance 100 contained in the specimen binds to the primary antibody 101 immobilized on the solid phase 20 .
  • the control unit 72 drives the second driving device 50 to remove the solid phase 20 from the first well 31 as shown in FIG. 9(C) (step S13 in FIG. 7).
  • the control unit 72 drives the first driving device 40 to move the container 30 to a position where the solid phase 20 faces the second well 32 as shown in FIGS. 10(A) and 11(A). Rotate (step S14 in FIG. 7).
  • the control unit 72 drives the second driving device 50 to perform the immersion treatment for the second well 32 (step S15 in FIG. 7). Specifically, the controller 72 immerses the solid phase 20 in the second well 32 for a predetermined period of time as shown in FIG. is removed from the second well 32 .
  • control unit 72 controls the immersion and removal of the solid phase 20 shown in FIGS. It may be repeated multiple times.
  • the controller 72 drives the first driving device 40 to rotate the container 30 to the position where the solid phase 20 faces the third well 33 as shown in FIG. step S16).
  • the controller 72 drives the second driving device 50 to perform the immersion treatment for the third well 33 (step S17 in FIG. 7).
  • the immersion treatment in step S17 is the same as or similar to the immersion treatment in step S15. Since the solid phase 20 is added to the detection antibody reagent in the third well 33 by the processing of step S17, the detection antibody 102 binds to the target substance 100 immobilized on the solid phase 20.
  • the controller 72 drives the first driving device 40 to rotate the container 30 to the position where the solid phase 20 faces the fourth well 34 as shown in FIG. step S18).
  • the controller 72 drives the second driving device 50 to perform the immersion treatment for the fourth well 34 (step S19 in FIG. 7).
  • the immersion treatment in step S19 is the same as or similar to the immersion treatment in step S15. Since the solid phase 20 is put into the washing liquid of the fourth well 34 by the processing of step S19, the detection antibody reagent adhering to the solid phase 20 is washed off.
  • the controller 72 drives the first driving device 40 to rotate the container 30 to the position where the solid phase 20 faces the fifth well 35 as shown in FIG. step S20).
  • the controller 72 drives the second driving device 50 to perform the immersion treatment for the fifth well 35 (step S21 in FIG. 7).
  • the immersion treatment in step S21 is the same as or similar to the immersion treatment in step S15. Since the solid phase 20 is added to the labeled antibody reagent in the fifth well 35 by the processing in step S21, the enzyme-labeled antibody 103 binds to the detection antibody 102 immobilized on the solid phase 20.
  • the controller 72 drives the first driving device 40 to rotate the container 30 to the position where the solid phase 20 faces the sixth well 36 as shown in FIG. step S22).
  • the control unit 72 drives the second driving device 50 to perform the immersion treatment for the sixth well 36 (step S23 in FIG. 7).
  • the immersion treatment in step S23 is the same as or similar to the immersion treatment in step S15. Since the solid phase 20 is put into the washing liquid of the sixth well 36 by the processing of step S23, the labeled antibody reagent adhering to the solid phase 20 is washed off.
  • the controller 72 drives the first driving device 40 to rotate the container 30 to the position where the solid phase 20 faces the seventh well 37 as shown in FIG. step S24).
  • the controller 72 drives the second driving device 50 to perform the immersion treatment for the seventh well 37 (step S25 in FIG. 7).
  • the immersion treatment in step S25 is the same as or similar to the immersion treatment in step S15. Since the solid phase 20 is added to the substrate reagent in the seventh well 37 by the processing in step S24, the enzymatic activity of the enzyme-labeled antibody 103 immobilized on the solid phase 20 causes the dye to be generated from the surface of the solid phase 20. It spreads to substrate reagents. The substrate reagent is therefore colored.
  • the substrate reagent is agitated by moving the solid phase 20 up and down with respect to the substrate reagent, so that the entire color of the substrate reagent can be homogenized in a short period of time.
  • the solid phase 20 may be further displaced in the lateral direction relative to the substrate reagent.
  • a method of laterally displacing the solid phase 20 relative to the substrate reagent for example, a method of slightly rotating the container 30 alternately clockwise and counterclockwise relative to the solid phase 20. can be used.
  • the substrate reagent Based on the coloring state of the substrate reagent thus obtained, it is possible to confirm whether or not the target substance 100 is contained in the specimen, and if the target substance 100 is contained, its concentration. For example, when the target substance 100 is contained in the sample, the substrate reagent develops color, so whether or not the sample contains the target substance 100 can be confirmed by the presence or absence of color development. In addition, the higher the concentration of the target substance 100 contained in the sample, the more intensely the substrate reagent develops color. Therefore, the concentration of the target substance 100 in the sample can be confirmed based on the intensity or absorbance of the color development.
  • the sample analyzer 10 comprises a solid phase 20, a container 30, and driving devices 40,50.
  • a specific binding reagent that specifically binds to the target substance contained in the specimen is immobilized on the solid phase 20 .
  • the container 30 is provided with a plurality of wells 31 to 37 in which specimens, detection reagents used for detection of target substances, and washing solutions are individually placed.
  • the driving devices 40 and 50 move the solid phase 20 and the container 30, respectively, to immerse and remove the solid phase 20 from each of the plurality of wells 31-37.
  • the specimen analysis method of this embodiment includes the step of immobilizing a specific binding reagent on the solid phase 20, and the specimen, detection reagent, and washing solution are individually placed in a plurality of wells 31 to 37 provided in the container 30. and immersing and removing the solid phase 20 into and out of each of the plurality of wells 31-37 by moving the solid phase 20 and the container 30 with the driving devices 40,50.
  • the solid phase 20 is immersed in and removed from each of the plurality of wells 31 to 37 by the driving devices 40 and 50, so manual work by the operator is almost unnecessary.
  • the sample analyzer 10 of the present embodiment has a simple configuration, it is possible to reduce the price.
  • the sample analyzer 10 of the present embodiment is easily available, it can be used in various situations according to user needs, such as analyzing samples at home or remotely. It becomes possible to realize an analysis environment.
  • the container 30 is formed in a disc shape around the axis m11.
  • a plurality of wells 31 to 37 are arranged side by side in the circumferential direction A of the container 30 . According to this configuration, the relative positions of the plurality of wells 31 to 37 and the solid phase 20 can be easily changed by rotating the container 30 around the axis m11.
  • the container 30 includes a well 31 containing a sample, a well 32 containing a washing solution, a well 33 containing a detection antibody reagent, a well 34 containing a washing solution, a well 35 containing a labeled antibody reagent, and a well 36 containing a washing solution. , and a well 37 in which a substrate reagent is placed are provided in the circumferential direction A in this order. According to this configuration, the same or similar analysis as the indirect sandwich method of the ELISA (Enzyme-Linked Immunosorbent Assay) method can be automatically performed.
  • ELISA Enzyme-Linked Immunosorbent Assay
  • the sample analyzer 10 has a first driving device 40 that rotates the container 30 about the axis m11, and a solid phase 20 that reciprocates in a direction parallel to the axis m11, thereby moving each of the plurality of wells 31 to 37. and a second driving device 50 for dipping and removing the solid phase 20 . According to this configuration, it is possible to easily realize a configuration that allows the solid phase 20 to be immersed in and taken out from the plurality of wells 31 to 37 .
  • the sample analyzer 10 further includes a control section 72 that controls the first driving device 40 and the second driving device 50 .
  • the control unit 72 drives the first driving device 40 to perform position control for matching the rotational position of the container 30 to a predetermined initial position, and then drives the first driving device 40 and the second driving device 50 to rotate a plurality of An immersion control for immersing and removing the solid phase 20 is performed for each of the wells 31 to 37 of .
  • An immersion control for immersing and removing the solid phase 20 is performed for each of the wells 31 to 37 of .
  • a projecting portion 39 is formed on the outer peripheral surface of the container 30 .
  • the sample analyzer 10 further includes a position detection section 71 that detects the position of the projecting section 39 .
  • the control unit 72 performs position control to match the rotational position of the container 30 with the initial position based on the position of the protrusion 39 detected by the position detection unit 71 . According to this configuration, the rotational position of the container 30 can be easily matched with the initial position.
  • a detection reagent containing an enzyme-labeled detection antibody is used as the detection reagent placed in the well 33 .
  • the well 35 is filled with a substrate reagent. In this case, wells 36 and 37 are unnecessary.
  • the target substance detection method includes the direct antibody method, the indirect antibody method, the chemiluminescent enzyme immunoassay method (CLEIA), the chemiluminescence immunoassay method (CLIA), and the indirect fluorescence method. Any detection method such as an antibody method (IIF; indirect immunofluorescence) can be used.
  • the label is not limited to an enzyme, and any label such as a fluorescent dye, a quantum dot, or a radioisotope can be used.
  • a substance other than the primary antibody 101 may be used.
  • the novel coronavirus spike protein may be used instead of the primary antibody 101 .
  • a container 30 shown in FIG. 14 is used in the sample analyzer 10 of the present embodiment.
  • the container 30 of this embodiment includes a rotating body 330 and multiple tubes 340 .
  • the rotating body 330 is formed in a disc shape around the axis m11.
  • a plurality of gripping portions 331 capable of gripping the tube 340 are formed at equal angular intervals on the outer periphery of the rotating body 330 .
  • the rotating body 330 is detachably attached to the rotating portion 41 of the first driving device 40 . Therefore, when the motor device 42 of the first driving device 40 rotates the rotating portion 41, the rotating body 330 rotates integrally with the rotating portion 41 about the axis m11.
  • tube 340 has an opening 341 at one end.
  • a plurality of tubes 340 contain any of a specimen, a detection antibody reagent, a labeled antibody reagent, a substrate reagent, and a wash solution.
  • the solid phase 20 is immersed and removed via an opening 341 .
  • the plurality of tubes 340 can be removed from the rotating body 330, so maintenance of the portion where samples and the like are put is facilitated.
  • the sample analyzer 10 of the third embodiment further includes an imaging device 80, a display device 81, and an irradiation device .
  • the imaging device 80 captures an image of the substrate reagent contained in the well 37 to acquire image data thereof, and transmits the acquired image data of the substrate reagent to the control device 70 .
  • the imaging device 80 corresponds to the imaging section.
  • the imaging target of the imaging device 80 is the substrate reagent contained in the well 37 .
  • the display device 81 is a device that displays the analysis result as an image, characters, or the like.
  • a liquid crystal display, an organic EL (Electro Luminescence), a touch panel, or the like can be used as the display device 81.
  • the irradiation device 82 is a device that irradiates the substrate reagent contained in the well 37 with light.
  • the sample analyzer 10 of this embodiment is installed, for example, in a dark room. Therefore, when the imaging device 80 acquires the image data of the substrate reagent while the well 37 is irradiated with light from the irradiation device 82, the image data in which only the substrate reagent is emphasized, in other words, the objects around the well 37 It is possible to acquire image data in which the image is not reflected as much as possible.
  • the control device 70 further has an image analysis section 73 and a display section 74 as functional components realized by the processor executing a program stored in the storage device.
  • the image analysis unit 73 determines the degree of enzymatic activity of the solid phase 20 by image recognition based on the image data acquired by the imaging device 80 . Specifically, after the solid phase 20 has been immersed in and taken out of the well 37, the image analysis unit 73 captures the image data of the substrate reagent contained in the well 37 after the solid phase 20 has been taken out. The presence or absence of the target substance 100 in the sample and the concentration of the target substance 100 are detected by acquiring the image data from the sample 80 and analyzing the acquired image data.
  • the image analysis unit 73 determines whether the sample contains the target substance 100 based on the color of the substrate reagent. For example, the image analysis unit 73 determines whether the sample contains the target substance 100 by comparing the RGB values of the substrate reagent with a predetermined threshold value. The predetermined threshold value is set according to the coloration of the enzyme-labeled antibody 103 due to its enzymatic activity. Further, when the image analysis unit 73 determines that the sample contains the target substance 100, the image analysis unit 73 determines the degree of color development of the target substance 100 by comparing the RGB values of the substrate reagent with a plurality of threshold values. At the same time, the concentration of the target substance 100 is detected according to the determined degree of color development.
  • the display unit 74 displays the analysis result of the image analysis unit 73 , specifically the presence or absence of the target substance 100 in the specimen and the concentration of the target substance 100 on the display device 81 .
  • the imaging device 80 and the image analysis section 73 function as an enzyme activity detection section that detects the degree of enzymatic activity of the enzyme-labeled antibody 103 with respect to the enzyme substrate. According to this configuration, the degree of enzymatic activity can be automatically detected, so convenience can be improved.
  • the sample analyzer 10 of the fourth embodiment will be described. The following description focuses on the differences from the sample analyzer 10 of the first embodiment.
  • the sample analyzer 10 of the present embodiment differs from the sample analyzer 10 of the first embodiment in that the gear structure of the rack gear 55 and the pinion gear 56 is used instead of the linear rail 512. different.
  • the rack gear 55 is arranged to extend in the vertical direction Z.
  • the rack gear 55 is supported so as to be relatively displaceable in the vertical direction Z with respect to the support portion 54 .
  • a grip portion 520 is fixed to the lower portion of the rack gear 55 . Therefore, the grip portion 520 is displaced in the vertical direction Z together with the rack gear 55 .
  • the grip portion 520 grips the proximal end portion of the mounting member 21 .
  • the proximal end of the solid phase 20 is attached to the attachment member 21 .
  • the pinion gear 56 is meshed with the rack gear 55.
  • the pinion gear 56 rotates clockwise indicated by arrow D1 and counterclockwise indicated by arrow D2 based on power transmitted from the motor device 53 .
  • the rack gear 55 is displaced in the vertical direction Z by rotating the pinion gear 56 . Specifically, when the pinion gear 56 rotates clockwise D1, the rack gear 55 is displaced upward Z1, and when the pinion gear 56 rotates counterclockwise D2, the rack gear 55 is displaced downward Z2.
  • a plurality of wells 321 and recesses 322 are formed on the outer edge of the container 30 .
  • a plurality of wells 321 contain detection antibody reagents, labeled antibody reagents, substrate reagents, washing solutions, and the like.
  • a sampling device 350 is assembled in the recess 322 .
  • a sample such as blood is put in the sampling instrument 350 .
  • the sample analyzer 10 further includes a columnar portion 90 that contacts the projecting portion 39 of the container 30 when the container 30 rotates about the axis m11, and fixing portions 91 and 92 for fixing the imaging device 80.
  • the sampling instrument 350 is formed in a substantially cylindrical shape with a bottom.
  • a flange portion 351 is formed on the upper portion of the sampling instrument 350 .
  • the bottom surface of the flange portion 351 and the upper surface 300 of the container 30 are brought into contact with each other in the vertical direction Z, so that the sampling instrument 250 is supported with respect to the container 30 at the position shown in FIG. there is
  • annular recess 360 is formed in the bottom surface of the container 30 .
  • annular convex portion 410 is formed on the top surface of the rotating portion 41 .
  • the circular convex portion 410 of the rotating portion 41 is fitted into the circular concave portion 360 of the container 30 .
  • the motor device 42 After aligning the initial positions of the rotating part 41 and the solid phase 20 as described above, the motor device 42 is driven to rotate the rotating part 41, and the motor device 53 is driven to move the solid phase 20 in the vertical direction Z.
  • the solid phase 20 is immersed in and taken out of the specimen of the collection instrument 350, and the solid phase 20 is immersed in and taken out of the detection antibody reagent, labeling antibody reagent, substrate reagent, and washing liquid of each of the plurality of wells 321.
  • the container 30 shown in FIG. 24 is used in the sample analyzer 10 of this modified example. Instead of the wells 321, the container 30 is formed with a plurality of concave grips 323 to which the tubes 340 of the second embodiment can be attached. Even with such a configuration, it is possible to obtain actions and effects similar to those of the sample analyzer 10 of the fourth embodiment.
  • FIG. 25 shows an enlarged structure around one well 130 out of a plurality of wells formed in the container 30 of this embodiment.
  • a recess 131 is formed in the upper surface 300 of the container 30 so as to surround the opening of the well 130 .
  • the recess 131 is formed to extend annularly around the central axis m21 of the well 130 .
  • the recess 131 opens at the upper surface 300 of the container 30 .
  • FIG. 26 shows an enlarged structure around the tip of the solid phase 20 of this embodiment.
  • the intermediate portion of the solid phase 20 is provided with a flange portion 22 .
  • the flange portion 22 is formed in a cylindrical shape around the central axis m22 of the solid phase 20.
  • An insertion hole 220 is formed through the flange portion 22 along its central axis. A solid phase 20 is inserted into the insertion hole 220 .
  • the tip of the solid phase 20 passes through the flange portion 22 and is exposed below the flange portion 22 .
  • a convex portion 221 is formed on the bottom surface 222 of the flange portion 22 .
  • the convex portion 221 is formed along the outer edge of the bottom surface 222 of the flange portion 22 .
  • the convex portion 221 is formed to extend annularly around the central axis m22.
  • the convex portion 211 formed on the bottom surface 222 of the flange portion 22 is It fits into the recess 131 formed around the opening of the well 130 in the container 30 .
  • the recess 131 is formed around the opening of the well 130 in the container 30 .
  • the solid phase 20 is formed with a convex portion 221 that fits into the concave portion 131 of the container 30 when the solid phase 20 is immersed in the well 130 . According to this configuration, as shown in FIG.
  • the tip of the solid phase 20 when the tip of the solid phase 20 is immersed in the well 130, the central axis m21 of the well 130 and the central axis m22 of the solid phase 20 substantially coincide. .
  • the tip of the solid phase 20 can be immersed substantially in the center of the well 130, so that the tip of the solid phase 20 can be more reliably immersed in the specimen, various reagents, washing liquid, etc. contained in the well 130. can be immersed in Therefore, it is possible to improve analysis accuracy.
  • a rotating body 330 is provided with a plurality of tubes 342a, 342b, 343a, 343b, 344a, 344b, 345a, 345b, 346a, 346b, 347a, 347b, 348a and 348b are mounted.
  • a pair of tubes 342a and 342b contain, for example, specimens.
  • a pair of tubes 344a and 344b contain, for example, detection antibody reagents, respectively.
  • a pair of tubes 346a and 346b contain, for example, a labeled antibody reagent.
  • a pair of tubes 348a and 348b contain, for example, a substrate reagent.
  • the pair of tubes 343a, 343b, the pair of tubes 345a, 345b, and the pair of tubes 347a, 347b contain, for example, cleaning liquid.
  • the first driving device 40 rotates the container 30 to dispose the pair of tubes 342a and 342b below the pair of solid phases 20a and 20b, respectively.
  • the second driving device 50 displaces the pair of solid phases 20a and 20b downward Z2 and upward Z1, thereby immersing and removing the pair of solid phases 20a and 20b into and out of the pair of tubes 342a and 342b, respectively. conduct.
  • the rotation of the container 30 by the first driving device 40 and the displacement of the pair of solid phases 20a and 20b by the second driving device 50 are repeatedly performed, whereby the pair of tubes 343a and 343b and the pair of tubes 344a and 344b are repeatedly performed.
  • sample analyzer 10 of this embodiment for example, different samples can be analyzed simultaneously by putting different samples into the pair of tubes 342a and 342b respectively. Therefore, it is possible to improve the analysis efficiency, for example, compared to the configuration having only one solid phase 20, such as the sample analyzer 10 of the fourth embodiment.
  • the sample analyzer 10 of the seventh embodiment is obtained by applying the imaging device 80 and the irradiation device 82 of the third embodiment to the sample analyzer 10 of the sixth embodiment.
  • a plate-like support member 83 is provided above the portion of the container 30 through which the tubes 348a and 348b pass.
  • there is 30 and 31 illustrate the case where the tube 348a is positioned below the support member 83.
  • the support member 83 is fixed at the position shown in FIGS. 30 and 31 by a strut or the like (not shown).
  • the imaging device 80 and the irradiation device 82 are provided on the bottom surface 832 of the support member 83 facing the top surface 300 of the container 30 .
  • the imaging device 80 is, for example, a CCD camera or a CMOS camera.
  • the irradiation device 82 is, for example, an LED light. In this embodiment, the irradiation device 82 corresponds to the irradiation section.
  • a side wall portion of the support member 83 is provided with a cap portion 830 .
  • the cap portion 830 has a function of preventing an external object from being reflected in the substrate reagent contained in the tube 348a, for example.
  • the tube 348a and the tube 348b are positioned below Z2 of the imaging device 80 in this order.
  • the imaging device 80 acquires image data of the substrate reagent contained in the tube 348a by imaging the tube 348a when the tube 348a is positioned downward.
  • the imaging device 80 acquires image data of the substrate reagent contained in the tube 348b by imaging the tube 348b when the tube 348b is positioned downward.
  • the light emitted from the irradiation device 82 is reflected by the upper surface 300 of the container 30, then reflected by the cap 830, and directed toward the opening 841 of the tube 348a as shown in FIG.
  • the bottom surface 830a of cap 830 which is exposed to the surface, may be reflected in the substrate reagent in tube 348a.
  • the cap portion 830 will appear in the image data captured by the imaging device 80 .
  • the color of the substrate reagent is determined based on the image data, the color of the cap 830 reflected in the substrate reagent becomes a disturbance, and the color of the substrate reagent, specifically, the RGB values of the substrate reagent is accurately determined. It may not be detected well.
  • the bottom surface 830a of the cap 830 is coated with a low-reflection material or non-reflection material that makes it difficult for light to be reflected.
  • a low-reflection material or non-reflection material that makes it difficult for light to be reflected.
  • light is less likely to be reflected on the bottom surface 830a of the cap portion 830, so that the cap portion 830 is less likely to be reflected in the substrate reagent described above. Therefore, it becomes possible to detect the color of the substrate reagent with higher accuracy.
  • the bottom surface 830 a of the cap portion 830 corresponds to the surface of the support member 83 facing the container 30 .
  • the tubes 348a and 348b are made of a transparent material, the color of the rotating body 330 may be reflected in the substrate reagent inside the tubes 348a and 348b. In order to avoid this, it is preferable to form the tubes 348a and 348b from a non-transparent material or to apply a non-transparent paint to the tubes 348a and 348b.
  • the container 30 is shaped like a rectangular parallelepiped.
  • the container 30 is, for example, a 96-well plate made of polypropylene, polystyrene, or the like.
  • the container 30 has a longitudinal direction in the direction indicated by arrow X and a lateral direction in the direction indicated by arrow Y.
  • the direction indicated by arrow X will be referred to as "transverse direction X”
  • the direction indicated by arrow Y will be referred to as "longitudinal direction Y”.
  • the lateral direction X, longitudinal direction Y, and vertical direction Z are orthogonal to each other.
  • the lateral direction X corresponds to the first direction
  • the longitudinal direction Y corresponds to the second direction
  • the vertical direction corresponds to the third direction.
  • Well groups 141 to 148 are arranged side by side in the longitudinal direction Y in the container 30 at predetermined intervals. Each of the well groups 141 to 148 is composed of eight wells arranged side by side in the lateral direction X at predetermined intervals. In this embodiment, the well groups 141 to 148 correspond to the accommodation section group.
  • Each well of the well group 141 contains, for example, a specimen.
  • Each well of well group 143 contains, for example, a detection antibody reagent.
  • Each well of well group 145 contains, for example, a labeled antibody reagent.
  • Each well of well group 147 contains, for example, a substrate reagent.
  • Each well of well group 142, each well of well group 144, and each well of well group 146 contains, for example, a washing solution.
  • a rack gear 150 is integrally provided on the bottom surface of the container 30 .
  • a pinion gear 43 is meshed with the rack gear 150 .
  • the first driving device 40 can reciprocate the container 30 in the longitudinal direction Y by rotating the pinion gear 43 with the motor device 42 .
  • the grip portion 520 is formed so as to extend in the transverse direction X.
  • the gripping portion 520 grips eight mounting members 21 that are arranged side by side in the transverse direction X with a predetermined interval therebetween.
  • a solid phase 20 is attached to the tip of each attachment member 21 .
  • a solid phase group 200 is composed of eight solid phases 20 mounted on eight mounting members 21 respectively.
  • the solid phase group 200 is arranged above the container 30 .
  • the first driving device 40 displaces the container 30 in the longitudinal direction Y to displace the plurality of wells of the well group 141 below the plurality of solid phases 20 of the solid phase group 200 respectively. position each.
  • the second driving device 50 displaces the solid phase group 200 downward Z2 and upward Z1, thereby immersing and removing the plurality of solid phases 20 of the solid phase group 200 in each of the plurality of wells of the well group 141. I do.
  • each well of the well group 142 displacement of the container 30 in the longitudinal direction Y by the first driving device 40 and displacement of the solid-phase group 200 in the vertical direction Z by the second driving device 50 are repeatedly performed, whereby each well of the well group 142, Each solid phase of solid phase group 200 is dipped into and removed from each well of well group 143, each well of well group 144, each well of well group 145, each well of well group 146, and each well of well group 147 in order. done.
  • sample analyzer 10 of this embodiment for example, by putting different samples into each well of the well group 141, those different samples can be analyzed simultaneously. Therefore, it is possible to improve the analysis efficiency, for example, compared to the configuration having only one solid phase 20, such as the sample analyzer 10 of the fourth embodiment.
  • the said embodiment can also be implemented in the following forms, for example.
  • the shape of the solid phase 20 can be arbitrarily changed, such as a square prism shape.
  • the concave portion 131 may be formed in the flange portion 22 of the solid phase 20 and the convex portion 221 may be formed around the opening of the well 130 of the container 30 .
  • the configuration of the first driving device 40 can be arbitrarily changed as long as the container 30 can be rotated.
  • the configuration of the second driving device 50 can be arbitrarily changed as long as the solid phase 20 can be displaced in the vertical direction Z.
  • the marking portion formed on the container 30 is not limited to the projecting portion 39, and any configuration that can detect the rotational position of the container 30, such as a concave portion, can be used.
  • the sample analyzer 10 is not limited to moving both the solid phase 20 and the container 30. For example, by moving only the solid phase 20, the solid phase 20 is immersed and It may take out. Further, the sample analyzer 10 may immerse and take out the solid phase 20 from the plurality of wells 31 to 37 by moving only the container 30 . In short, the sample analyzer 10 should only immerse and remove the solid phase 20 from the plurality of wells 31 to 37 and the plurality of tubes 340 by moving at least one of the solid phase 20 and the container 30. .
  • the sample analyzer 10 of each embodiment may be operable by a personal computer, smart phone, or the like.
  • an appropriate method can be adopted, and for example, remote operation using wireless communication can be used.
  • the configuration of the above embodiment can be applied to any detection method as long as it uses a reagent that specifically reacts.
  • the configuration of the above embodiment can be used for a method of detecting a nucleic acid using complementary binding of bases, a method of detecting an enzyme using an enzyme-substrate reaction, or the like.

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Abstract

Un analyseur d'échantillon (10) comprend une phase solide (20), un récipient (30) et des dispositifs d'entraînement (40 et 50). Un réactif de liaison spécifique qui se lie spécifiquement à une substance cible contenue dans un échantillon est immobilisé sur la phase solide (20). Le récipient (30) est pourvu d'une pluralité de puits (31-37) dans laquelle des échantillons, des réactifs de détection utilisés pour la détection de substances cibles et des solutions de lavage sont placés individuellement. Les dispositifs d'entraînement (40 et 50) déplacent la phase solide (20) et le récipient (30) pour immerger et retirer la phase solide (20) dans et à partir de chacun de la pluralité de puits (31-37).
PCT/JP2022/043035 2021-11-22 2022-11-21 Analyseur d'échantillon et procédé d'analyse d'échantillon WO2023090446A1 (fr)

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JPH08506893A (ja) * 1993-02-01 1996-07-23 ラブシステムズ オユ サンプル用ウェルの形状と一致する担体を用いる固相イムノアッセイ
JP2009053125A (ja) * 2007-08-29 2009-03-12 Hitachi High-Technologies Corp 自動分析装置
JP2014501933A (ja) * 2011-01-08 2014-01-23 アクセス メディカル システムズ，リミティド 免疫学的検定試験のためのシステム
JP2017187496A (ja) * 2016-04-08 2017-10-12 アイセンス，インコーポレーテッド 遠心分離が可能な円形タイプカートリッジおよびこれを用いたモジュール式自動分析装置
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JPH08506893A (ja) * 1993-02-01 1996-07-23 ラブシステムズ オユ サンプル用ウェルの形状と一致する担体を用いる固相イムノアッセイ
JP2009053125A (ja) * 2007-08-29 2009-03-12 Hitachi High-Technologies Corp 自動分析装置
JP2014501933A (ja) * 2011-01-08 2014-01-23 アクセス メディカル システムズ，リミティド 免疫学的検定試験のためのシステム
JP2017187496A (ja) * 2016-04-08 2017-10-12 アイセンス，インコーポレーテッド 遠心分離が可能な円形タイプカートリッジおよびこれを用いたモジュール式自動分析装置
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