WO2024070614A1 - Récipient de test, dispositif d'amplification d'acide nucléique et procédé de test d'amplification d'acide nucléique - Google Patents

Récipient de test, dispositif d'amplification d'acide nucléique et procédé de test d'amplification d'acide nucléique Download PDF

Info

Publication number
WO2024070614A1
WO2024070614A1 PCT/JP2023/032960 JP2023032960W WO2024070614A1 WO 2024070614 A1 WO2024070614 A1 WO 2024070614A1 JP 2023032960 W JP2023032960 W JP 2023032960W WO 2024070614 A1 WO2024070614 A1 WO 2024070614A1
Authority
WO
WIPO (PCT)
Prior art keywords
flow path
sample
nucleic acid
acid amplification
temperature side
Prior art date
Application number
PCT/JP2023/032960
Other languages
English (en)
Japanese (ja)
Inventor
隆次 清水
薫 重松
誠司 大西
薫 上之園
Original Assignee
Ｐｈｃホールディングス株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ｐｈｃホールディングス株式会社 filed Critical Ｐｈｃホールディングス株式会社
Publication of WO2024070614A1 publication Critical patent/WO2024070614A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

Definitions

  • the present invention relates to a test container, a nucleic acid amplification device, and a nucleic acid amplification test method.
  • a nucleic acid amplification device that performs a so-called thermal cycle to cause a polymerase chain reaction (PCR) in nucleic acids (genes) such as DNA (Deoxyribonucleic Acid) through a thermal denaturation step, an annealing step, and an extension reaction step, thereby amplifying the nucleic acid.
  • PCR polymerase chain reaction
  • nucleic acid amplification device that is equipped with a mechanism for amplifying nucleic acids as well as a detection unit for detecting nucleic acids, and is capable of detecting amplified nucleic acids in real time (see, for example, Patent Document 1).
  • a nucleic acid amplification device is called a real-time PCR device.
  • the real-time PCR device amplifies the nucleic acid by controlling the temperature of the reaction sample, which contains the nucleic acid to be tested as well as test reagents such as primers and fluorescent probes.
  • the real-time PCR device then irradiates the reaction sample with excitation light that excites a fluorescent dye, and quantitatively measures the amplified nucleic acid based on the fluorescence generated by the fluorescent dye.
  • a thermal cycle including a thermal denaturation step, an annealing step, and an extension reaction step is repeated multiple times to amplify nucleic acids.
  • the present invention was made in consideration of these circumstances, and aims to provide a test container, a nucleic acid amplification device, and a nucleic acid amplification test method that can shorten the time required for testing.
  • test container used in a nucleic acid amplification test performed in a nucleic acid amplification device, A substrate;
  • a flow channel is provided on the substrate and accommodates a sample containing nucleic acid;
  • the flow path is a thermal flow path having a circulation flow path through which the sample circulates; and a pump provided in the circulation flow path for moving the sample.
  • a nucleic acid amplification device that performs a nucleic acid amplification test in a state in which a test container having a circulation flow path through which a sample circulates is incorporated, a pump driving unit that drives a pump provided in the circulation flow path; a low-temperature heater section that heats a low-temperature side heat section of the circulation flow path to a first predetermined temperature; a high-temperature heater section that heats a high-temperature side heat section of the circulation flow path to a second predetermined temperature that is higher than the first predetermined temperature; and a fluorescence detection unit that detects the fluorescence of a fluorescent dye contained in the sample.
  • the nucleic acid amplification test method is to A nucleic acid amplification test method performed in a nucleic acid amplification device that performs a nucleic acid amplification test in a state in which a test container having a flow path in which a sample is contained is attached, comprising:
  • the flow path is a thermal flow path including a circulation flow path through which a sample circulates during a nucleic acid amplification test; a dispensing channel connected to the thermal channel; a pump provided in the circulation flow path for moving the sample; a first valve provided in the thermal flow path between the circulation flow path and the dispensing flow path; A second valve provided in the circulation flow path,
  • the nucleic acid amplification test method is performing a dispensing process in which the pump is driven while the first valve is open and the second valve is closed to move the sample from the dispensing flow path to the circulation flow path; and performing a thermal cycle process of heating the sample while circulating it in the circulation channel by driving the pump with the first
  • the present invention provides a test container, a nucleic acid amplification device, and a nucleic acid amplification test method that can reduce the time required for testing.
  • FIG. 1 is a plan view of a container for inspection according to a first embodiment.
  • FIG. 2 is a bottom view of the container for inspection.
  • FIG. 3 is a schematic cross-sectional view of the first high temperature side valve, the first low temperature side valve, and the second valve.
  • FIG. 4 is a cross-sectional perspective view showing an example of a pump.
  • FIG. 5 is an exploded perspective view showing a first modified example of the pump.
  • FIG. 6 is a cross-sectional perspective view showing a first modified example of the pump.
  • FIG. 7 is a cross-sectional perspective view showing a second modified example of the pump.
  • FIG. 8 is a schematic cross-sectional view of the nucleic acid amplification device according to the first embodiment.
  • FIG. 1 is a plan view of a container for inspection according to a first embodiment.
  • FIG. 2 is a bottom view of the container for inspection.
  • FIG. 3 is a schematic cross-sectional view of the first high temperature side valve, the
  • FIG. 9 is a perspective view for explaining the configuration of the pump drive unit.
  • FIG. 10 is a perspective view for explaining the configuration of a rotating member of the pump drive unit.
  • FIG. 11 is a schematic diagram for explaining the operation of the pump driving unit.
  • FIG. 12 is a schematic diagram showing the configuration of a fluorescence detection device.
  • FIG. 13 is a time chart showing the temperature cycle in a nucleic acid amplification test.
  • FIG. 14A is a diagram for explaining the state of the cartridge in a nucleic acid amplification test.
  • FIG. 14B is a diagram for explaining the state of the cartridge in a nucleic acid amplification test.
  • FIG. 14C is a diagram for explaining the state of the cartridge in a nucleic acid amplification test.
  • FIG. 14A is a diagram for explaining the state of the cartridge in a nucleic acid amplification test.
  • FIG. 14B is a diagram for explaining the state of the cartridge in a nucleic acid amplification
  • FIG. 14D is a diagram for explaining the state of the cartridge in a nucleic acid amplification test.
  • FIG. 15 is a schematic cross-sectional view of the nucleic acid amplification device according to the second embodiment.
  • FIG. 16 is a perspective view of a pump driver of the nucleic acid amplification apparatus according to the third embodiment.
  • FIG. 17 is a cross-sectional perspective view of the pump drive unit.
  • FIG. 18 is a schematic diagram for explaining the operation of the pump driving unit.
  • test container The test container, nucleic acid amplification device, and nucleic acid amplification test method according to the present invention will be described below with reference to the drawings. Note that the same components are given the same reference numerals.
  • FIGS. 1 to 14D A container for testing, a nucleic acid amplification device, and a nucleic acid amplification testing method according to a first embodiment of the present invention will be described with reference to FIGS. 1 to 14D.
  • FIG. 1 is a plan view of a cartridge C according to a first embodiment of the present invention.
  • FIG. 2 is a bottom view of the cartridge C.
  • the cartridge C is an example of a test container.
  • Cartridge C is used in a nucleic acid amplification test carried out in a nucleic acid amplification device such as a PCR device or a real-time PCR device.
  • a nucleic acid amplification device such as a PCR device or a real-time PCR device.
  • Cartridge C is set in the nucleic acid amplifier 6 while containing a reaction sample (hereinafter simply referred to as "sample”).
  • sample includes, for example, the nucleic acid (hereinafter also referred to as "target nucleic acid”) of the test subject (e.g., a virus) and test reagents such as primers and fluorescent probes.
  • target nucleic acid the nucleic acid
  • a thermal cycle process is performed, thereby carrying out processes such as a thermal denaturation process, an annealing process, and an extension reaction process.
  • the target nucleic acid is amplified in cartridge C.
  • the cartridge C has a base portion 1 and a flow path 2.
  • the base part 1 is made of a colorless and transparent synthetic resin and has a rectangular plate shape.
  • the synthetic resin that constitutes the base part 1 is, for example, polypropylene, polycarbonate, or cycloolefin polymer.
  • elements such as a first low-temperature side valve 30, a first high-temperature side valve 31, a pump 34, and a second valve 37 that constitute the flow path 2 described below are fixed to the base part 1.
  • the base part 1 and each of these elements are made by a molding method such as two-color molding.
  • the upper surface (front surface) of the base part 1 (cartridge C) is the first main surface.
  • the lower surface (rear surface) of the base part 1 (cartridge C) is the second main surface.
  • the first and second main surfaces of the base part 1 (cartridge C) are a pair of opposing surfaces.
  • the base portion 1 has a high temperature region R1, a low temperature region R2, and an intermediate region R3.
  • the high-temperature region R1 is a region that is heated by a high-temperature heater unit 641 of the nucleic acid amplifier 6 described below during a nucleic acid amplification test (hereinafter also simply referred to as "during the test"). Specifically, the high-temperature region R1 is a region that includes the right end portion of the base portion 1.
  • the directions indicated by the arrows labeled with up and down characters in each figure are described as up or down for convenience.
  • the directions indicated by the arrows labeled with left and right characters in each figure are described as right or left for convenience.
  • the left and right direction of the base part 1 is also the longitudinal direction of the base part 1.
  • the directions indicated by the arrows labeled with front and back characters in each figure are described as front or back for convenience.
  • the front and back direction of the base part 1 is also the lateral direction of the base part 1.
  • the directions indicated by the arrows labeled with up and down characters in each figure may or may not match the vertical direction when the cartridge C is set in the nucleic acid amplification device.
  • the position of the high temperature region is not limited to the position of the high temperature region R1 in this embodiment.
  • the position of the high temperature region may be set appropriately in relation to the position of the high temperature heater unit 641 of the nucleic acid amplification device 6.
  • the high temperature region R1 has a first high temperature region R11 and a second high temperature region R12.
  • the first high temperature region R11 is a portion including the right end portion of the base portion 1.
  • the second high temperature region R12 is a region to the left of the second high temperature region R12 (the side closer to the low temperature region R2).
  • the first high temperature region R11 is an area that is heated by the first high temperature heater 641a of the high temperature heater section 641 during inspection.
  • the second high temperature region R12 is an area that is heated by the second high temperature heater 641b of the high temperature heater section 641 during inspection. During inspection, the temperature of the second high temperature region R12 becomes higher than the temperature of the first high temperature region R11.
  • the low-temperature region R2 is an area that is heated by the low-temperature heater 642a of the nucleic acid amplifier 6 during testing. Specifically, the low-temperature region R2 is an area that includes the left end of the base portion 1.
  • the position of the low temperature region is not limited to the position of the low temperature region R2 in this embodiment.
  • the position of the low temperature region may be set appropriately depending on its relationship with the position of the low temperature heater 642a of the nucleic acid amplification device 6.
  • the intermediate region R3 is a region provided between the high-temperature region R1 and the low-temperature region R2 in the left-right direction of the base portion 1.
  • the intermediate region R3 may be considered as a portion that does not face the high-temperature heater portion 641 and the low-temperature heater portion 642 of the nucleic acid amplifier 6 during testing.
  • the flow path 2 is composed of a groove formed on the lower surface (second main surface) of the base part 1, and a number of elements fixed to the base part 1.
  • the groove constituting the flow path 2 opens downward.
  • the lower opening of the groove is blocked by a film-like sealing member 50 (see Figure 4) fixed to the lower surface of the base part 1.
  • the flow path 2 has a dispensing flow path 2a and thermal flow paths 3a, 3b, 3c, 3d, and 3e.
  • the dispensing flow path 2a is a flow path through which the sample moves during the dispensing process in the test.
  • the dispensing flow path 2a has a sample storage section 20 and multiple (five in this embodiment) individual reagent holding sections 21.
  • the sample storage section 20 is provided in the high temperature region R1 (specifically, the first high temperature region R11) of the base section 1.
  • the sample storage section 20 has a sample storage space 20a, a sample dropping port 20b, and an air hole 20c.
  • the sample storage space 20a is a space surrounded by a long groove in the front-rear direction formed on the underside of the base part 1.
  • the sample storage space 20a is a space in which the sample dropped into the sample dropping port 20b by the operator is temporarily stored.
  • the sample storage space 20a is connected to the thermal channels 3a to 3e via channel elements L1 and L2.
  • the channel elements are elements that make up the channel 2, and are spaces defined by grooves formed on the underside of the base 1.
  • the first end (rear end in this embodiment) of the sample storage space 20a is connected to the low temperature ends of the thermal channels 3a to 3e via channel element L1.
  • the second end (front end in this embodiment) of the sample storage space 20a is connected to the high temperature ends of the thermal channels 3a to 3e via channel element L2.
  • the sample inlet 20b and the air hole 20c are each formed by a through hole provided in the base portion 1.
  • the sample inlet 20b and the air hole 20c each communicate between the sample storage space 20a and the external space.
  • the openings of the sample inlet 20b and the air hole 20c on the external space side are blocked by film-like sealing members 51, 52 (see Figure 1) during testing.
  • the individual reagent carrying section 21 is an example of a reagent carrying section, and is provided at the left end of the base section 1 shown in FIG. 2. In other words, the individual reagent carrying section 21 is provided between the left end of the base section 1 and the low-temperature side heat section 33 of the thermal flow paths 3a to 3e described below.
  • the individual reagent carrying section 21 is provided in an area that is not heated by the heater section 64 of the nucleic acid amplification device 6 (is not easily affected by the heat of the heater section 64).
  • the individual reagent holding section 21 is connected to the sample storage section 20 via the flow path element L1.
  • the individual reagent holding section 21 holds a reagent (hereinafter referred to as an "individual reagent") in advance.
  • the individual reagents are reagents that correspond to the target nucleic acid.
  • the individual reagents include, for example, a dried enzyme (e.g., DNA polymerase), a primer, and a probe having a fluorescent dye and a quencher.
  • the type of the individual reagent may be appropriately determined depending on the target nucleic acid.
  • the individual reagent may also be a liquid.
  • the flow path 2 may have one common reagent carrying section 22 between the sample storage section 20 and the individual reagent carrying section 21.
  • a reagent (hereinafter referred to as a "common reagent") is carried in advance in the common reagent carrying section 22.
  • the common reagent is a reagent common to all the target nucleic acids targeted by the thermal flow paths 3a to 3e.
  • the common reagent may be carried in the individual reagent carrying section 21. In this case, the common reagent carrying section 22 may be omitted.
  • the thermal flow paths 3a to 3e are arranged in parallel to one another and are flow paths through which the sample moves during thermal cycle processing in testing.
  • the thermal flow paths 3a to 3e are arranged in a line in a predetermined direction (in this embodiment, the front-to-back direction) in the base portion 1. All of the thermal flow paths 3a to 3e can be considered collectively as the thermal flow path.
  • the thermal flow paths 3a to 3e are parallel flow paths having the same configuration.
  • the flow path 2 has five thermal flow paths 3a to 3e.
  • the number of thermal flow paths may be considered as the number of test items.
  • thermal flow path 3a The configuration of thermal flow path 3a will be described below.
  • the configuration of thermal flow paths 3b to 3e can be understood by appropriately interpreting the description of the configuration of thermal flow path 3a. Note that the same reference numerals are used for the configurations common to thermal flow paths 3a to 3e.
  • Thermal flow path 3a is a flow path provided in correspondence with individual reagent holder 21. Specifically, thermal flow path 3a has a first low-temperature side valve 30, a first high-temperature side valve 31, and a circulation flow path 32.
  • the circulation flow path 32 is sandwiched between the first low-temperature side valve 30 and the first high-temperature side valve 31. When the first low-temperature side valve 30 and the first high-temperature side valve 31 are closed, the circulation flow path 32 becomes a closed flow path.
  • the specific configurations of the first low-temperature side valve 30, the first high-temperature side valve 31, and the circulation flow path 32 are described below.
  • the first low-temperature side valve 30 is an example of a first valve, and is a valve fixed to the base portion 1.
  • the first low-temperature side valve 30 is provided between the dispensing flow path 2a and the thermal flow path 3a.
  • the first low-temperature side valve 30 is provided between the individual reagent holding section 21 and the circulation flow path 32. Specifically, the first low-temperature side valve 30 is provided in the flow path element L3 that connects the individual reagent holding section 21 and the circulation flow path 32.
  • the flow path element L3 corresponds to an example of a first connecting flow path.
  • the first low-temperature side valve 30 is provided in the low-temperature region R2 of the base part 1.
  • the portion of the base part 1 to which the first low-temperature side valve 30 is fixed is a through-hole that passes through the base part 1 in the vertical direction.
  • the state of the first low-temperature side valve 30 is controlled by the nucleic acid amplifier 6 during testing. Specifically, the first low-temperature side valve 30 is switched between an open state and a closed state by the first low-temperature side valve drive unit 651 (see FIG. 8) of the nucleic acid amplifier 6.
  • the sample and air are permitted to pass through the first low-temperature side valve 30.
  • the first low-temperature side valve 30 is closed, the sample and air are prohibited from passing through the first low-temperature side valve 30.
  • FIG. 3 is a schematic cross-sectional view of the first cold side valve 30.
  • the first cold side valve 30 is made of an elastic material that is elastic and heat resistant.
  • the elastic material is, for example, a thermoplastic elastomer (TPE).
  • TPE thermoplastic elastomer
  • the first cold side valve 30 has a main body portion 30a, a valve flow path 30b, and a pressed portion 30c.
  • the main body 30a is substantially rectangular and is fixed to the base 1.
  • the valve flow path 30b is a tunnel-shaped space defined by a groove formed on the underside of the main body 30a.
  • the pressed portion 30c is a portion that is pressed downward by the first low-temperature side valve driving unit 651 (see FIG. 8) of the nucleic acid amplification device 6 during testing.
  • the pressed portion 30c is configured as a convex portion provided on the upper surface of the main body portion 30a.
  • the structure of the pressed portion 30c is not limited to a convex portion.
  • the configuration of the pressed portion 30c may be a concave shape.
  • the valve flow path 30b is open.
  • the first low-temperature side valve 30 is in the open state.
  • the sample and air are allowed to pass through the valve flow path 30b.
  • the pressed portion 30c When the pressed portion 30c is pressed downward by the first low-temperature side valve drive portion 651 (hereinafter referred to as the "pressed state of the pressed portion 30c"), the groove that defines the valve flow path 30b elastically deforms, and the valve flow path 30b is closed.
  • the first low-temperature side valve 30 When the pressed portion 30c is in the pressed state, the first low-temperature side valve 30 is in the closed state.
  • the sample and air are prohibited from passing through the valve flow path 30b.
  • the first low-temperature side valve 30 is in a closed state when pressed by the first low-temperature side valve drive unit 651.
  • the first low-temperature side valve 30 opens due to its own restoring force.
  • the first low-temperature side valves 30 of the thermal channels 3a to 3e are connected to each other by the connection parts 30d on the upper surface of the base part 1, as shown in FIG. 1. This configuration contributes to improving the manufacturing work efficiency of the cartridge C.
  • the first high temperature side valve 31 is a valve fixed to the base portion 1, and is provided between the sample storage portion 20 and the circulation flow path 32. Specifically, the first high temperature side valve 31 is provided in the flow path element L4 that connects the sample storage portion 20 and the circulation flow path 32.
  • the flow path element L4 corresponds to an example of a second connection flow path.
  • the flow path element L4 is connected to the flow path element L2.
  • the first high temperature side valve 31 is provided in the high temperature region R1 of the base part 1.
  • the portion of the base part 1 where the first high temperature side valve 31 is fixed is a through hole that passes through the base part 1 in the vertical direction.
  • the state of the first high temperature side valve 31 is controlled by the nucleic acid amplifier 6 during testing. Specifically, the first high temperature side valve 31 is switched between an open state and a closed state by the first high temperature side valve drive unit 652 (see FIG. 8) of the nucleic acid amplifier 6.
  • the sample and air are permitted to pass through the first high temperature side valve 31.
  • the first high temperature side valve 31 is closed, the sample and air are prohibited from passing through the first high temperature side valve 31.
  • FIG. 3 is also a schematic cross-sectional view of the first high temperature side valve 31.
  • the first high temperature side valve 31 is made of an elastic material that is elastic and heat resistant.
  • the elastic material is, for example, a thermoplastic elastomer (TPE).
  • TPE thermoplastic elastomer
  • the first high temperature side valve 31 has a main body portion 31a, a valve flow path 31b, and a pressed portion 31c.
  • the configurations of the main body 31a, the valve flow path 31b, and the pressed portion 31c are similar to the configurations of the main body 30a, the valve flow path 30b, and the pressed portion 30c of the first low-temperature side valve 30.
  • the pressed portion 31c is the portion that is pressed downward by the first high temperature side valve driving portion 652 (see FIG. 8) of the nucleic acid amplifier 6 during testing.
  • valve flow path 31b is open.
  • the first high temperature side valve 31 When the pressed portion 31c is not pressed, the first high temperature side valve 31 is open. When the first high temperature side valve 31 is open, the sample and air are allowed to pass through the valve flow path 31b.
  • the groove that defines the valve flow path 31b elastically deforms and the valve flow path 31b closes.
  • the first high-temperature side valve 31 is in the closed state.
  • the sample and air are prohibited from passing through the valve flow path 31b.
  • the first high temperature side valve 31 is in a closed state when it is pressed by the first high temperature side valve drive unit 652. Then, when the pressure from the first high temperature side valve drive unit 652 is released, the first high temperature side valve 31 opens due to its own restoring force.
  • the first high temperature side valves 31 of the thermal flow paths 3a to 3e are connected to each other by the connection part 31d on the upper surface of the base part 1, as shown in FIG. 1. This configuration contributes to improving the manufacturing work efficiency of the cartridge C.
  • the circulation flow path 32 is a flow path through which the sample circulates during the thermal cycle process in the test.
  • the circulation flow path 32 is a flow path provided between the first low-temperature side valve 30 and the first high-temperature side valve 31. Such a circulation flow path 32 becomes a closed flow path when the first low-temperature side valve 30 is closed and the first high-temperature side valve 31 is closed.
  • the circulation flow path 32 is composed of a first parallel flow path 32a and a second parallel flow path 32b whose ends are connected to each other.
  • a flow path element L3 is also connected to the position where the first end (the left end in this embodiment) of the first parallel flow path 32a and the first end (the left end in this embodiment) of the second parallel flow path 32b are connected.
  • a flow path element L4 is connected to the position where the second end (the right end in this embodiment) of the first parallel flow path 32a and the second end (the right end in this embodiment) of the second parallel flow path 32b are connected.
  • the circulation flow path 32 has a low-temperature side heating section 33, a pump 34, a preheating section 35, a high-temperature side heating section 36, and a second valve 37.
  • the sample moves cyclically through the circulation flow path 32 in a predetermined direction (the direction indicated by the arrow Y1 in FIG. 2).
  • This predetermined direction is also referred to as the circulation direction.
  • upstream and downstream refer to the upstream and downstream sides in the circulation direction.
  • the pump 34, preheat section 35, high-temperature side heat section 36, and second valve 37 are arranged in this order from the upstream side in the circulation direction.
  • the specific configurations of the low-temperature side heat section 33, pump 34, preheat section 35, high-temperature side heat section 36, and second valve 37 will be described below.
  • the low-temperature side heat section 33 is provided in the low-temperature region R2 of the base section 1. In other words, the low-temperature side heat section 33 is provided at the first end (the left end in this embodiment) of the first parallel flow path 32a.
  • the low-temperature side heat section 33 is a space defined by a groove formed on the underside of the base section 1.
  • the low-temperature side heat section 33 is a serpentine space that is folded back at least once (twice in this embodiment).
  • the folding angle of the low-temperature side heat section 33 is 180 degrees. Note that the folding angle and number of folding times of the low-temperature side heat section 33 are not limited to those of this embodiment.
  • the low-temperature side heat section 33 is heated by the low-temperature heater section 642 (specifically, the low-temperature heater 642a) of the nucleic acid amplifier 6 during testing.
  • the heating temperature of the low-temperature heater section 642 is the first predetermined temperature T1.
  • the predetermined area including the low-temperature side heat section 33 in the cartridge C is the area to be detected (hereinafter referred to as the "detection area") when the amplification results of the nucleic acid are determined by the fluorescence detection device 7 described below.
  • a detection area is set for each of the thermal flow paths 3a to 3e. Therefore, the cartridge C has five detection areas.
  • the pump 34 is a member for moving the sample in the flow path 2.
  • the pump 34 is provided downstream of the low-temperature side heat section 33 in the sample circulation direction. In other words, the pump 34 is provided in the intermediate region R3 of the base section 1. In further other words, the pump 34 is provided downstream of the low-temperature side heat section 33 in the first parallel flow path 32a.
  • the portion of the base 1 where the pump 34 is fixed is a through hole that passes through the base 1 in the vertical direction.
  • the pump 34 and the low-temperature side heat section 33 are connected by a flow path element L5.
  • FIG. 4 is a cross-sectional perspective view of the pump 34.
  • the pump 34 is made of an elastic material that is elastic and heat-resistant.
  • the elastic material is, for example, a thermoplastic elastomer (TPE).
  • TPE thermoplastic elastomer
  • the pump 34 has a main body portion 34a and a pump space forming portion 34b.
  • the main body 34a is a generally rectangular plate and is fixed to the base 1.
  • the pump space forming portion 34b is provided on the main body 34a.
  • the pump space forming portion 34b extends in a predetermined direction (left-right direction in this embodiment) and is configured with a protrusion that is concave on the lower surface and convex on the upper surface.
  • the tunnel-shaped space defined by the lower surface of the pump space forming portion 34b is the pump space 34c.
  • the lower opening of the pump space forming portion 34b is blocked by a sealing member 50.
  • the first end (upstream end) of the pump space 34c is connected to the flow path element L5.
  • the second end (downstream end) of the pump space 34c is connected to the flow path element L6.
  • the pump space 34c, the flow path element L5, and the flow path element L6 are located at the same height.
  • the pump space 34c, the flow path element L5, and the flow path element L6 are connected in a straight line.
  • the pumps 34 of the thermal flow paths 3a to 3e are connected to each other by the connection part 34d on the upper surface of the base part 1, as shown in FIG. 1. This configuration contributes to improving the manufacturing work efficiency of the cartridge C.
  • the pump 34 as described above is driven by the pump drive unit 661 of the nucleic acid amplification device 6.
  • the operation of the pump 34 will be described later together with an explanation of the pump drive unit 661.
  • the pump 34 is made of a single, integrally molded material.
  • the pump may be made of a combination of multiple components, as shown in Figures 5 and 6.
  • FIGS. 5 and 6 are diagrams showing a pump 34A according to the first modified example.
  • the pump 34A has an upper pump space forming member 34e and a lower pump space forming member 34f.
  • the upper pump space forming member 34e and the lower pump space forming member 34f are each fixed to the base portion 1.
  • the upper pump space forming member 34e is fixed to the upper surface of the base part 1.
  • the lower pump space forming member 34f is fixed to the lower surface of the base part 1, facing the upper pump space forming member 34e in the vertical direction.
  • the upper pump space forming member 34e is a protruding member that extends in a predetermined direction (left-right direction in this embodiment) and has a concave lower surface and a convex upper surface.
  • the upper pump space forming member 34e is made of an elastic material that is elastic and heat resistant.
  • the elastic material is, for example, a thermoplastic elastomer (TPE).
  • the lower pump space forming member 34f has a base 34g and a protrusion 34h.
  • the base 34g is a plate-shaped member that extends in a predetermined direction (in this embodiment, the left-right direction).
  • the base 34g is fixed to the base portion 1.
  • the protrusion 34h extends in a predetermined direction (left and right in this embodiment) and is located at the center in the width direction of the upper surface of the base 34g.
  • the cross-sectional shape of the protrusion 34h is approximately rectangular. Note that the cross-section refers to the cross section of the protrusion 34h cut by a plane perpendicular to the extension direction of the protrusion 34h.
  • the lower pump space forming member 34f is made of the same synthetic resin as the base portion 1.
  • the synthetic resin constituting the lower pump space forming member 34f is, for example, polypropylene, polycarbonate, or cycloolefin polymer.
  • the lower pump space forming member 34f does not have to be colorless and transparent, and may be colored.
  • pump space 34i the space defined by the lower surface of upper pump space forming member 34e and the upper surface of protrusion portion 34h of lower pump space forming member 34f is pump space 34i.
  • the first end (upstream end) of the pump space 34i is connected to the flow path element L5 (see FIG. 2).
  • the second end (downstream end) of the pump space 34i is connected to the flow path element L6 (see FIG. 2).
  • the pump space 34i is located at a higher position than the flow path elements L5 and L6. Therefore, the pump space 34i is connected to the flow path elements L5 and L6 in a stepped manner.
  • FIG. 7 is a cross-sectional perspective view of pump 34B according to variant example 2.
  • Pump 34B is a tubular member made of an elastic material having elasticity and heat resistance.
  • the elastic material may be, for example, a thermoplastic elastomer (TPE) or a silicone resin.
  • Both ends of the pump 34B are fixed to a pair of pump fixing parts 11a provided on the base part 1. Note that in FIG. 7, only the pump fixing part 11a on one side (left side) is shown, and the pump fixing part 11a on the other side (right side) is omitted.
  • pump space 34j In the case of pump 34B, the space defined by the inner circumferential surface of pump 34B is pump space 34j. A first end (upstream end) of pump space 34j is connected to flow path element L5 (see FIG. 2). A second end (downstream end) of pump space 34j is connected to flow path element L6 (see FIG. 2).
  • the pump space 34j is located at a higher position than the flow path elements L5 and L6. Therefore, the pump space 34j, the flow path elements L5 and L6 are connected in a stepped manner via a pair of pump fixing parts 11a.
  • All of the pumps 34, 34A, and 34B described above are driven by the pump drive unit 661 of the nucleic acid amplifier 6.
  • the configuration of the pump drive unit 661 of the nucleic acid amplifier 6 is determined appropriately depending on the configuration of the pumps 34, 34A, and 34B.
  • the preheat section 35 raises the temperature of the sample circulating through the circulation flow path 32 to a predetermined temperature before the sample flows into the high-temperature side heat section 36.
  • the preheating section 35 is provided downstream of the pump 34 in the direction of sample circulation. In other words, the preheating section 35 is provided in the high-temperature region R1 (specifically, the second high-temperature region R12) of the base section 1.
  • the preheat section 35 is provided downstream of the pump 34 in the first parallel flow path 32a.
  • the preheat section 35 is connected to the pump 34 via the flow path element L6.
  • the preheat section 35 is also provided upstream of the high-temperature side heat section 36, which will be described later.
  • the preheat section 35 is a space defined by a groove formed on the underside of the base section 1.
  • the preheat section 35 is a serpentine space that is folded back at least once (once in this embodiment).
  • the folding angle of the preheat section 35 is 180 degrees. Note that the folding angle and number of folding times of the preheat section 35 are not limited to those of this embodiment.
  • the preheat section 35 is heated by the second high-temperature heater 641b of the nucleic acid amplifier 6 during testing.
  • the heating temperature of the second high-temperature heater 641b is a third predetermined temperature T3.
  • the third predetermined temperature T3 is higher than the first predetermined temperature T1, which is the heating temperature of the low-temperature heater 642a, and the second predetermined temperature T2, which is the heating temperature of the first high-temperature heater 641a.
  • the third predetermined temperature T3 is 1 to 3 degrees higher than the second predetermined temperature T2 (T2+1 degree ⁇ T3 ⁇ T2+3 degrees).
  • the preheat section 35 described above can increase the temperature of the sample as it flows into the high-temperature side heat section 36. This can shorten the time it takes for the sample to reach the second predetermined temperature T2 in the high-temperature side heat section 36 during testing. As a result, the time required for the thermal cycle process during testing can be shortened, and therefore the time required for testing can be shortened.
  • the high-temperature side heat section 36 is provided in the high-temperature region R1 (specifically, the first high-temperature region R11) of the base section 1. In other words, the high-temperature side heat section 36 is provided at the second end (the right end in this embodiment) of the second parallel flow path 32b.
  • the high-temperature side heat section 36 is connected to the preheat section 35 via the flow path element L7.
  • the high-temperature side heat section 36 is a space defined by a groove formed on the underside of the base section 1.
  • the high-temperature side heat section 36 is a serpentine space that is folded back at least once (twice in this embodiment).
  • the fold angle of the high-temperature side heat section 36 is 180 degrees. Note that the fold angle and number of folds of the high-temperature side heat section 36 are not limited to those of this embodiment.
  • the high-temperature side heating section 36 is heated by the first high-temperature heater 641a of the nucleic acid amplification device 6 during testing.
  • the heating temperature of the first high-temperature heater 641a is a second predetermined temperature T2.
  • the second predetermined temperature T2 is higher than the first predetermined temperature T1, which is the heating temperature of the low-temperature heater section 642, and lower than the third predetermined temperature T3, which is the heating temperature of the second high-temperature heater 641b.
  • the second predetermined temperature T2 is, for example, 90 degrees or higher and 97 degrees or lower.
  • the second valve 37 is a valve fixed to the base portion 1 and is provided in the flow path element L8 that connects the low-temperature side heat portion 33 and the high-temperature side heat portion 36.
  • the second valve 37 is provided in the intermediate region R3 of the base portion 1.
  • the second valve 37 is provided downstream of the high-temperature side heat portion 36 in the second parallel flow path 32b.
  • the portion of the base portion 1 where the second valve 37 is fixed is a through hole that passes through the base portion 1 in the vertical direction.
  • the operation of the second valve 37 is controlled by the nucleic acid amplifier 6 during testing. Specifically, the second valve 37 is switched between an open state and a closed state by a second valve drive unit 653 (see FIG. 8) of the nucleic acid amplifier 6.
  • FIG. 3 is also a schematic cross-sectional view of the second valve 37.
  • the second valve 37 is made of an elastic material that is elastic and heat-resistant.
  • the elastic material is, for example, a thermoplastic elastomer (TPE).
  • TPE thermoplastic elastomer
  • the second valve 37 has a main body portion 37a, a valve flow path 37b, and a pressed portion 37c.
  • the configurations of the main body 37a, the valve flow path 37b, and the pressed portion 37c are similar to the configurations of the main body 30a, the valve flow path 30b, and the pressed portion 30c of the first low-temperature side valve 30.
  • the pressed portion 37c is the portion that is pressed downward by the second valve driving portion 653 (see FIG. 8) of the nucleic acid amplification device 6 during testing.
  • valve flow path 37b is open.
  • the second valve 37 When the pressed portion 37c is not pressed, the second valve 37 is open. When the second valve 37 is open, the sample and air are allowed to pass through the valve flow path 37b.
  • the pressed portion 37c When the pressed portion 37c is pressed downward by the second valve drive portion 653 (hereinafter referred to as the "pressed state of the pressed portion 37c"), the groove that defines the valve flow path 37b elastically deforms, and the valve flow path 37b is closed.
  • the second valve 37 When the pressed portion 37c is pressed, the second valve 37 is closed. When the second valve 37 is closed, the sample and air are prohibited from passing through the valve flow path 37b.
  • the second valve 37 is in a closed state when pressed by the second valve drive unit 653.
  • the second valve 37 opens due to its own restoring force.
  • the second valves 37 of the thermal channels 3a to 3e are connected to each other by the connection portion 37d on the upper surface of the base portion 1, as shown in FIG. 1. This configuration contributes to improving the efficiency of the manufacturing process of the cartridge C.
  • thermal flow path 3a has been described above, but flow path 2 has thermal flow paths 3b to 3e that have a configuration similar to that of thermal flow path 3a.
  • Thermal flow paths 3a to 3e are arranged side by side in the front-to-rear direction as shown in FIG. 2.
  • Each of the thermal channels 3a to 3e has an individual reagent carrying portion 21. And, each of the individual reagent carrying portions 21 of the thermal channels 3a to 3e is pre-loaded with an individual reagent corresponding to the target nucleic acid.
  • nucleic acid amplification tests for multiple types of target nucleic acids can be performed simultaneously with a single cartridge C.
  • the pre-processing step of mixing the sample and the individual reagent outside the cartridge can be omitted.
  • the time required for one test can be shortened while reducing the workload of the operator.
  • the nucleic acid amplification test is carried out with the cartridge C set in the nucleic acid amplifier 6.
  • the configuration of the nucleic acid amplifier 6 is described below.
  • the nucleic acid amplification device 6 has a housing 60, a control unit 61, a cartridge support unit 62, a vibration unit 63, a heater unit 64, a valve drive unit 65, a liquid delivery unit 66, a sample position detection unit 67, and a fluorescence detection device 7.
  • FIG. 8 also shows a schematic diagram of cartridge C.
  • the arrangement of each element of cartridge C shown in FIG. 8 is slightly different from the arrangement of each element of cartridge C shown in FIG. 2.
  • the housing 60 is box-shaped and has a storage space 601.
  • the storage space 601 contains the elements 61 to 67 and 7 that make up the nucleic acid amplification device 6.
  • the control unit 61 is supported by the housing 60 and controls the overall operation of the nucleic acid amplification device 6.
  • the control unit 61 may be configured in such a way that a CPU, ROM, RAM, and HDD are connected via a bus, or may be configured as a one-chip LSI, etc.
  • the control unit 61 has a first control unit 611 and a second control unit 612.
  • the first control unit 611 controls the operation of the vibration unit 63, the valve drive unit 65, the liquid delivery unit 66, the sample position detection unit 67, and the fluorescence detection device 7.
  • the second control unit 612 controls the operation of the heater unit 64. Specifically, the second control unit 612 controls the operation of the first high-temperature heater 641a, the second high-temperature heater 641b, and the low-temperature heater 642a of the heater unit 64.
  • the first control unit 611 and the second control unit 612 may be configured as a common control unit.
  • the function of the control unit 61 will be described later together with the operation of the nucleic acid amplification device 6 and the nucleic acid amplification test method.
  • the cartridge support portion 62 is an example of a container support portion, is supported by the housing 60, and is a member that supports the cartridge C.
  • the cartridge support portion 62 may include, for example, a table on which the cartridge C is placed, and a locking mechanism that secures the cartridge C to the table.
  • the cartridge support portion 62 supports the cartridge C vertically. In other words, when the cartridge C is supported by the cartridge support portion 62, the first main surface and the second main surface of the cartridge C face horizontally.
  • the cartridge C is attached to the cartridge support portion 62.
  • the cartridge support portion 62 may also be configured to support the cartridge C horizontally. In this case, when the cartridge C is supported by the cartridge support portion 62, the first main surface and the second main surface of the cartridge C face vertically.
  • the vibration unit 63 is supported by the housing 60. During the inspection, the vibration unit 63 applies vibration to the cartridge C supported by the cartridge support unit 62 under the control of the control unit 61.
  • the vibration unit 63 applies vibrations to the individual reagent holding portion 21 of the cartridge C during testing.
  • the vibration unit 63 is provided at a position where it can impart vibrations to the individual reagent holding portion 21 of the cartridge C during testing. Specifically, the vibration unit 63 is provided at a position facing the individual reagent holding portion 21 of the cartridge C in a predetermined direction (horizontal direction in this embodiment) during testing.
  • the vibration portion 63 applies vibration to the individual reagent holding portion 21, thereby promoting mixing of the sample and the individual reagent in the individual reagent holding portion 21.
  • the fact that the sample has been contained in the individual reagent holding portion 21 is detected by the sample position detection portion 67, which will be described later.
  • the vibration unit 63 has multiple (five in this embodiment) transducers 63a.
  • Each of the transducers 63a is composed of, for example, an ultrasonic transducer.
  • Each of the transducers 63a applies vibration to the individual reagent carriers 21 of the thermal channels 3a to 3e.
  • the vibration portion 63 may be omitted.
  • Another means is a means for moving the sample back and forth around the individual reagent holding portion 21 with the pump 34 centered on the individual reagent holding portion 21. The sample can be moved back and forth by reversing the driving direction of the pump 34. The fact that the sample has been contained in the individual reagent holding portion 21 is detected by the sample position detection portion 67 described below.
  • the heater unit 64 is supported by the housing 60. During inspection, the heater unit 64 heats the cartridge C under the control of the control unit 61. Specifically, the heater unit 64 has a low-temperature heater unit 642 and a high-temperature heater unit 641.
  • the low-temperature heater section 642 has a low-temperature heater 642a.
  • the low-temperature heater 642a heats the low-temperature region R2 of the cartridge C under the control of the control unit 61 (specifically, the second control unit 612). In other words, the low-temperature heater 642a heats the low-temperature side heat unit 33 of the flow path 2 in the cartridge C under the control of the control unit 61.
  • the low-temperature heater 642a constantly heats the low-temperature region R2 (specifically, the low-temperature side heat section 33) of the cartridge C during inspection.
  • the heating temperature of the low-temperature heater 642a is the first predetermined temperature T1.
  • the low-temperature heater 642a may be composed of multiple (five in this embodiment) low-temperature heaters provided for each low-temperature side heat section 33 of the thermal flow paths 3a to 3e, or may be composed of a single low-temperature heater.
  • the low-temperature heater 642a is provided at a position where it can heat the low-temperature region R2 (specifically, the low-temperature side heat section 33) of the cartridge C during inspection. Specifically, the low-temperature heater 642a is provided at a position facing the low-temperature region R2 (specifically, the low-temperature side heat section 33) of the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the high-temperature heater unit 641 heats the high-temperature region R1 of the cartridge C under the control of the control unit 61 (specifically, the second control unit 612).
  • the high-temperature heater unit 641 has a first high-temperature heater 641a and a second high-temperature heater 641b.
  • the first high-temperature heater 641a heats the first high-temperature region R11 of the cartridge C under the control of the control unit 61. In other words, the first high-temperature heater 641a heats the high-temperature side heat section 36 of the cartridge C under the control of the control unit 61. The first high-temperature heater 641a also heats the sample storage section 20 under the control of the control unit 61.
  • the first high-temperature heater 641a constantly heats the first high-temperature region R11 of the cartridge C during inspection.
  • the heating temperature of the first high-temperature heater 641a is a second predetermined temperature T2.
  • the second predetermined temperature T2 is higher than the first predetermined temperature, which is the heating temperature of the low-temperature heater 642a.
  • the first high-temperature heater 641a is supported at a position where it can heat the first high-temperature region R11 (specifically, the high-temperature side heat section 36 and the sample storage section 20) of the cartridge C during inspection. Specifically, the first high-temperature heater 641a is supported at a position facing the first high-temperature region R11 (specifically, the high-temperature side heat section 36 and the sample storage section 20) of the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the second high-temperature heater 641b is an example of a preheater, and heats the second high-temperature region R12 of the cartridge C under the control of the control unit 61. In other words, the second high-temperature heater 641b heats the preheat section 35 of the cartridge C under the control of the control unit 61.
  • the second high-temperature heater 641b constantly heats the second high-temperature region R12 (specifically, the preheat section 35) of the cartridge C during inspection.
  • the heating temperature of the second high-temperature heater 641b is a third predetermined temperature T3.
  • the third predetermined temperature T3 is higher than the first predetermined temperature T1, which is the heating temperature of the low-temperature heater 642a, and the second predetermined temperature T2, which is the heating temperature of the first high-temperature heater 641a.
  • the third predetermined temperature T3 is 1 to 3 degrees higher than the second predetermined temperature T2 (T2 + 1 degree ⁇ T3 ⁇ T2 + 3 degrees).
  • the third predetermined temperature T3 may be, for example, greater than or equal to 98 degrees and less than 100 degrees.
  • the second high-temperature heater 641b is supported at a position where it can heat the second high-temperature region R12 (specifically, the preheat section 35) of the cartridge C during inspection. Specifically, the second high-temperature heater 641b is supported at a position facing the second high-temperature region R12 (specifically, the preheat section 35) of the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the valve drive unit 65 is supported by the housing 60. During inspection, the valve drive unit 65 controls the open/close states of the first low-temperature side valve 30, the first high-temperature side valve 31, and the second valve 37 of the cartridge C under the control of the control unit 61 (specifically, the first control unit 611).
  • valve drive unit 65 has a first low-temperature side valve drive unit 651, a first high-temperature side valve drive unit 652, and a second valve drive unit 653.
  • the first low-temperature side valve drive unit 651 switches the first low-temperature side valve 30 of cartridge C between an open state and a closed state under the control of the control unit 61.
  • the first low-temperature side valve drive unit 651 is supported by the housing 60 and has a pressing unit 651a. During inspection, the pressing unit 651a moves toward and away from the first low-temperature side valve 30 based on the power of an actuator (not shown) such as an electric motor. The distance that the pressing unit 651a can move is called the movement stroke of the pressing unit 651a.
  • the pressing portion 651a is provided at a position where it can press the pressed portion 30c of the first low-temperature side valve 30 in the cartridge C during inspection. Specifically, the pressing portion 651a is supported at a position facing the pressed portion 30c of the first low-temperature side valve 30 in the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the pressing portion 651a When the pressing portion 651a has moved to the position closest to the first low-temperature side valve 30 (in other words, one end of the movement stroke), the pressing portion 651a presses the pressed portion 30c of the first low-temperature side valve 30.
  • the state in which the pressing portion 651a presses the pressed portion 30c is referred to as the pressing state of the first low-temperature side valve drive portion 651.
  • the valve flow path 30b of the first low-temperature side valve 30 is closed.
  • the first low-temperature side valve 30 is in a closed state.
  • the state in which the pressing portion 651a is not pressing the pressed portion 30c is referred to as the non-pressed state of the first low-temperature side valve drive portion 651.
  • the valve flow path 30b of the first low-temperature side valve 30 opens.
  • the first low-temperature side valve 30 is in an open state.
  • the first low-temperature side valve driving unit 651 may be composed of multiple valve driving units (five in this embodiment) provided for each of the first low-temperature side valves 30 of the thermal flow paths 3a to 3e, or may be composed of a single valve driving unit.
  • the control unit 61 controls the operation of each of the multiple valve driving units independently.
  • the first high temperature side valve drive unit 652 switches the first high temperature side valve 31 of cartridge C between an open state and a closed state under the control of the control unit 61 (specifically, the first control unit 611).
  • the first high temperature side valve drive unit 652 is supported by the housing 60 and has a pressing unit 652a.
  • the pressing unit 652a moves toward and away from the first high temperature side valve 31 based on the power of an actuator (not shown) such as an electric motor.
  • the distance that the pressing unit 652a can move is called the movement stroke of the pressing unit 652a.
  • the pressing portion 652a is provided at a position where it can press the pressed portion 31c of the first high temperature side valve 31 in the cartridge C during inspection.
  • the pressing portion 652a is provided at a position facing the pressed portion 31c of the first high temperature side valve 31 in the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the pressing portion 652a presses the pressed portion 31c of the first high-temperature side valve 31.
  • the state in which the pressing portion 652a presses the pressed portion 31c is referred to as the pressing state of the first high-temperature side valve drive portion 652.
  • the valve flow path 31b of the first high-temperature side valve 31 is closed.
  • the first high-temperature side valve 31 is in a closed state.
  • the state in which the pressing portion 652a is not pressing the pressed portion 31c is referred to as the non-pressed state of the first high-temperature side valve drive portion 652.
  • the valve flow path 31b of the first high-temperature side valve 31 opens.
  • the first high-temperature side valve 31 is in an open state.
  • the first high temperature side valve drive unit 652 may be composed of multiple valve drive units (five in this embodiment) provided for each of the first high temperature side valves 31 of the thermal flow paths 3a to 3e, or may be composed of a single valve drive unit.
  • the control unit 61 controls the operation of each of the multiple valve drive units independently.
  • the second valve drive unit 653 switches the second valve 37 of the cartridge C between an open state and a closed state under the control of the control unit 61 (specifically, the first control unit 611).
  • the second valve drive unit 653 is supported by the housing 60 and has a pressing unit 653a.
  • the pressing unit 653a moves toward and away from the second valve 37 based on the power of an actuator (not shown) such as an electric motor.
  • the distance that the pressing unit 653a can move is referred to as the movement stroke of the pressing unit 653a.
  • the pressing portion 653a is provided at a position where it can press the pressed portion 37c of the second valve 37 in the cartridge C during inspection. Specifically, the pressing portion 653a is provided at a position facing the pressed portion 37c of the second valve 37 in the cartridge C in a predetermined direction (horizontal direction in this embodiment) during inspection.
  • the pressing portion 653a presses the pressed portion 37c of the second valve 37.
  • the state in which the pressing portion 653a presses the pressed portion 37c is referred to as the pressing state of the second valve drive portion 653.
  • the valve flow path 37b of the second valve 37 is closed.
  • the second valve 37 is in the closed state.
  • the state in which the pressing portion 653a is not pressing the pressed portion 37c is referred to as the non-pressed state of the second valve drive portion 653.
  • the valve flow path 37b of the second valve 37 is open.
  • the second valve 37 is in an open state.
  • the second valve driving unit 653 may be composed of multiple valve driving units (five in this embodiment) provided for each second valve 37 of the thermal flow paths 3a to 3e, or may be composed of a single valve driving unit.
  • the control unit 61 controls the operation of each of the multiple valve driving units independently.
  • the liquid delivery unit 66 is supported by the housing 60.
  • the liquid delivery unit 66 moves the sample in the cartridge C during testing.
  • the liquid delivery unit 66 has a pump drive unit 661.
  • the pump drive unit 661 drives the pump 34 of the cartridge C under the control of the control unit 61 (specifically, the first control unit 611).
  • a pump drive unit 661 is provided for each pump 34 in the thermal flow paths 3a to 3e.
  • the pump drive unit 661 is made up of five pump drive units. The configuration of one pump drive unit 661 will be described below.
  • Figure 9 is a perspective view of the pump drive unit 661.
  • the pump drive unit 661 has a base 661a and a rotating member 661b.
  • Figure 10 is a perspective view of the pump drive unit 661 as viewed from below the base 661a.
  • FIG. 11 is a diagram for explaining the operation of the pump drive unit 661.
  • Thermal flow path 3f is shown in FIGS. 9 to 11.
  • the configuration of thermal flow path 3f differs from the configuration of thermal flow paths 3a to 3e.
  • the base 661a is fixed to the housing 60 via a fastening part (not shown) such as a bolt.
  • the base 661a also serves as a member that rotatably supports the rotating member 661b.
  • the base 661a has an upper base 661f and a lower base 661g. Only the upper base 661f of the base 661a is shown in FIG. 9. Only the lower base 661g of the base 661a is shown in FIG. 10.
  • the upper base 661f is fixed to the housing 60 via fastening parts such as bolts (not shown).
  • the lower base 661g is fixed to the underside of the upper base 661f.
  • the lower base 661g has a pair of rotational support parts 661c, 661d.
  • the rotational support parts 661c, 661d rotatably support both ends of the rotating member 661b.
  • the lower base 661g has a generally rectangular through hole 661h in the center.
  • the through hole 661h is positioned so as to face the pump 34 in the vertical direction.
  • the rotating member 661b is a member that drives the pump 34.
  • the rotating member 661b is a shaft member. Both ends of the rotating member 661b in the axial direction are supported by the rotation support parts 661c and 661d of the base part 661a.
  • the rotating member 661b is rotatable with respect to the base part 661a.
  • the rotating member 661b has a pressing portion 661e on its outer circumferential surface.
  • the pressing portion 661e is configured with a spiral ridge.
  • the pressing portion 661e is disposed in a through hole 661h in the lower base portion 661g. In this state, the pressing portion 661e faces the pump space forming portion 34b (see FIG. 4) of the pump 34 via the through hole 661h.
  • the rotating member 661b rotates based on the power of an actuator (not shown) such as an electric motor.
  • the pump drive unit 661 having the above-mentioned configuration is arranged so as to cover the pump space forming portion 34b (see FIG. 4) of the pump 34 from a predetermined direction (horizontal direction in this embodiment).
  • the pressing portion 661e abuts against the pump space forming portion 34b of the pump 34.
  • the pressing portion 661e crushes the pump space forming portion 34b at the position where the pressing portion 661e abuts against the pump space forming portion 34b.
  • Figure 11 is a schematic diagram showing a contact position D between the pressing portion 661e of the pump drive unit 661 and the pump space forming portion 34b of the pump 34.
  • the pressing portion 661e is omitted in Figure 11, at the position indicated by the contact position D, the pressing portion 661e presses against the pump space forming portion 34b of the pump 34.
  • the contact position D between the spiral pressing portion 661e and the pump space forming portion 34b moves in a predetermined direction (the direction indicated by the arrow Y2 in FIG. 11, which is the direction in which the sample circulates).
  • the pressing portion 661e applies a frictional force to the pump space forming portion 34b in the rotational direction of the rotating member 661b at the contact position D, thereby crushing the pump space forming portion 34b.
  • the direction of the frictional force is parallel to the main surface of the cartridge C and perpendicular to the extension direction of the pump space forming portion 34b.
  • the movement distance of the contact position D is referred to as the contact stroke of the pump drive portion 661.
  • the contact position D between the pressing portion 661e and the pump space forming portion 34b moves from one end of the contact stroke (the upstream end in the direction of circulation of the sample) to the other end of the contact stroke (the downstream end in the direction of circulation of the sample).
  • the contact position D between the pressing portion 661e and the pump space forming portion 34b repeatedly moves from one end of the contact stroke to the other end, so that the sample or air in the pump space 34c of the pump 34 is pumped in the circulation direction.
  • the sample or air in the pump space 34c moves, the sample and air in the flow path 2 move in conjunction with each other.
  • the direction of movement of the contact position D between the pressing portion 661e and the pump space forming portion 34b varies depending on the direction of rotation of the rotating member 661b. Therefore, by changing the direction of rotation of the rotating member 661b, the direction of movement of the sample and air in the flow channel 2 can be changed.
  • the pump drive unit 661 according to this embodiment is applicable to a pump having a pump space forming portion 34b that extends linearly, such as the pump 34 described above.
  • the configuration of the pump drive unit is not limited to the pump drive unit 661 according to this embodiment.
  • the configuration of the pump drive unit may be determined appropriately depending on the structure of the pump.
  • a pump driver 661 is provided for each pump 34 in the thermal flow paths 3a to 3e.
  • one pump driver 661 may be configured to drive pumps 34 in multiple (at least two) inspection flow paths among the thermal flow paths 3a to 3e.
  • the thermal flow channel 3b may be configured to be inverted in the front-rear direction.
  • the thermal flow channels 3a and 3b are in a line-symmetrical relationship with respect to a straight line parallel to the left-right direction.
  • the pump 34 of the thermal flow channel 3a and the pump 34 of the thermal flow channel 3b are arranged adjacent to each other in the front-rear direction, so that the pump 34 of the thermal flow channel 3a and the pump 34 of the thermal flow channel 3b can be easily driven by one pump driver 661.
  • the configuration of each of the thermal flow channels 3a to 3e may be set so that the function of each thermal flow channel is equivalent to that of the above-mentioned thermal flow channels 3a to 3e.
  • the number of pump drivers can be made smaller than the number of test flow channels, thereby reducing the number of parts constituting the nucleic acid amplification device.
  • the sample position detection unit 67 is supported by the housing 60.
  • the sample position detection unit 67 detects information regarding the position of the sample in the cartridge C during testing.
  • the information detected by the sample position detection unit 67 includes information indicating that the sample is contained in the low-temperature side heat unit 33 and information indicating that the sample is contained in the high-temperature side heat unit 36.
  • the information detected by the sample position detection unit 67 may also include information indicating that the sample is contained in the sample storage unit 20 and/or information indicating that the sample is contained in the individual reagent holder 21.
  • the sample position detection unit 67 has a low temperature side detection unit 671 and a high temperature side detection unit 672.
  • the low temperature side detection unit 671 and the high temperature side detection unit 672 are configured by separate detectors.
  • the low-temperature side detection section 671 is located at a position facing the low-temperature side heating section 33 of the cartridge C in a predetermined direction (horizontal in this embodiment) during inspection.
  • the low-temperature side detection unit 671 detects information indicating that a sample has been accommodated in the low-temperature side heat unit 33 (hereinafter referred to as "low-temperature side accommodation information").
  • the low-temperature side detection unit 671 is, for example, a detector that detects the low-temperature side accommodation information by optical processing (hereinafter referred to as “optical processing detector”).
  • the low-temperature side detection unit 671 may also be a detector that detects low-temperature side storage information by image processing (hereinafter referred to as an "image processing detector”).
  • image processing detector a detector that detects low-temperature side storage information by image processing
  • electrical processing detector a detector that detects low-temperature side storage information by electrical processing
  • the low-temperature side detection unit 671 detects the low-temperature side storage information based on the light transmittance and/or reflectance in the low-temperature side heat unit 33.
  • the low-temperature side detection unit 671 when the low-temperature side detection unit 671 is an optical processing detector, the low-temperature side detection unit 671 has a light source and a light receiving element.
  • the light source irradiates the low-temperature side heat unit 33.
  • the light receiving element receives light that has passed through the low-temperature side heat unit 33 and/or light that has been reflected by the low-temperature side heat unit 33.
  • the low-temperature side detection section 671 can detect low-temperature side containment information based on a change in the output of the light receiving element.
  • the low-temperature side detection unit 671 is an image detector, the low-temperature side detection unit 671 captures an image of the low-temperature side heat unit 33 using a camera, and acquires an image of the low-temperature side heat unit 33. The low-temperature side detection unit 671 then performs image analysis on the acquired image of the low-temperature side heat unit 33 to detect low-temperature side storage information.
  • the low-temperature side detection section 671 is an electrical detector, a pair of electrodes (not shown) is provided on the sealing member 50 of the cartridge C in a portion that covers the low-temperature side heat section 33.
  • the pair of electrodes is formed on the sealing member 50 by printing or the like.
  • the low-temperature side detection unit 671 also has a pair of terminals that are connected to a pair of electrodes of the sealing member 50 during testing. A voltage is applied to the pair of terminals. When no sample has flowed into the low-temperature side heat unit 33, no current flows between the pair of electrodes.
  • the low-temperature side detection section 671 can detect low-temperature side storage information based on the current flow state between the pair of electrodes.
  • the high-temperature side detection unit 672 detects information indicating that a sample has been contained in the high-temperature side heat unit 36 (hereinafter referred to as "high-temperature side containment information").
  • the high-temperature side detection unit 672 is provided at a position facing the high-temperature side heat unit 36 in the cartridge C in a predetermined direction (horizontal direction in this embodiment) during testing.
  • the rest of the configuration of the high-temperature side detection unit 672 is the same as the configuration of the low-temperature side detection unit 671.
  • the fluorescence detection device 7 is a device for detecting the fluorescence emitted from a fluorescent dye contained in a sample under the control of the control unit 61.
  • the fluorescent dye is bound to a probe in the sample together with a quencher that absorbs the fluorescence of the fluorescent dye.
  • the probe binds to the target nucleic acid together with the primer during the annealing step in the nucleic acid amplification test. Then, when the probe is decomposed during the extension reaction step in the nucleic acid amplification test, the fluorescent dye and quencher are separated.
  • the fluorescence detection device 7 detects the fluorescence of the fluorescent dye in the sample after the extension reaction step.
  • the control unit 61 determines the results of the amplification of the nucleic acid based on the detection results of the fluorescence detection device 7. The process in which the control unit 61 determines the results of the amplification of the nucleic acid is referred to as the amplification determination process.
  • the fluorescence detection device 7 is supported by a housing 60 and has a fluorescence detection unit 8 and a drive unit 9.
  • the fluorescence detection unit 8 is configured with an optical system including multiple optical elements. Specifically, the fluorescence detection unit 8 has a light-emitting optical system 80 and a light-receiving optical system 81.
  • the light-emitting optical system 80 is an optical system for irradiating the detection area of the cartridge C.
  • the detection area of the cartridge C is, for example, an area including the low-temperature side heat section 33.
  • the light emission optical system 80 has a light source unit 801, a filter unit 802, an object-side opening 820, and an objective lens 821.
  • light emitted from the light source unit 801 passes through the filter unit 802, the object-side opening 820, and the objective lens 821 and is irradiated onto the detection area of the cartridge C.
  • the light source unit 801 has at least one (two in this embodiment) light source 801a, 801b.
  • the light sources 801a, 801b are each fixed to a substrate 83 such as a printed circuit board.
  • the light sources 801a, 801b are each disposed at the front focal position of an objective lens 821, which will be described later.
  • Each of the light sources 801a and 801b is, for example, an LED light source, and emits white light or monochromatic light at least a part of which or the entirety of which is included in the excitation wavelength spectrum of the fluorescent dye.
  • the number of light sources is not limited to the number of light sources 801a and 801b in this embodiment. The number of light sources may be determined according to the number of types of fluorescent dyes contained in the sample.
  • the side closer to the cartridge C during inspection is referred to as the object side.
  • the side farther from the cartridge C during inspection is referred to as the image side.
  • the filter section 802 is provided on the object side of the light source section 801.
  • the filter section 802 has excitation filters 802a and 802b in a number corresponding to the number of light sources 801a and 801b.
  • the excitation filter 802a faces the light source 801a.
  • the excitation filter 802b faces the light source 801b.
  • Each of the excitation filters 802a and 802b is a bandpass filter that selectively passes light of a specific wavelength.
  • Each of the excitation filters 802a and 802b passes light of a wavelength that can excite the fluorescent dye contained in the sample and blocks light of other wavelengths.
  • excitation filter 802a passes light with a wavelength capable of exciting a first fluorescent dye contained in the sample.
  • excitation filter 802b passes light with a wavelength capable of exciting a second fluorescent dye contained in the sample. Therefore, the wavelength of light passing through excitation filter 802a is different from the wavelength of light passing through excitation filter 802b.
  • the light emitted by the light source 801a is incident on the excitation filter 802a. Then, light of a specific wavelength among the light incident on the excitation filter 802a passes through the excitation filter 802a.
  • the light emitted by the light source 801b is incident on the excitation filter 802b. Then, light of a specific wavelength among the light incident on the excitation filter 802b passes through the excitation filter 802b.
  • the object-side opening 820 is configured as a through hole provided in a plate-like member, and is provided on the object side of the filter section 802.
  • the object-side opening 820 adjusts the amount of light that passes through. Specifically, the object-side opening 820 passes a portion of the light that has passed through the filter section 802.
  • the object-side opening 820 is also an optical element that constitutes the light-receiving optical system 81.
  • the objective lens 821 is provided on the object side of the object-side opening 820.
  • the diameter of the objective lens 821 is larger than the inner diameter of the object-side opening 820. Therefore, all light that passes through the object-side opening 820 is incident on the objective lens 821.
  • the light sources 801a and 801b are positioned at the front focal position of the objective lens 821, so that the light that passes through the objective lens 821 is in the form of a parallel beam and is illuminated on the detection area of the cartridge C from any angle. As a result, the detection area of the cartridge C is illuminated with little unevenness in illumination.
  • the objective lens 821 is also an optical element that constitutes the light receiving optical system 81.
  • the light receiving optical system 81 is an optical system for detecting the fluorescence emitted by the fluorescent dye in the sample when determining the results of nucleic acid amplification.
  • the light receiving optical system 81 has, in order from the object side, an objective lens 821, an object side opening 820, a multi-fluorescence filter 810, an imaging lens 811, an image side opening 812, and a light receiving element 813.
  • the objective lens 821 is a lens shared with the light emission optical system 80. Fluorescence emitted by the fluorescent dye excited by the irradiation light from the light source unit 801 enters the objective lens 821 from the object side.
  • the light receiving optical system 81 is a so-called object-side telecentric optical system in which the chief ray ⁇ of the fluorescence incident on the objective lens 821 is parallel to the optical axis X of the objective lens 821.
  • the object-side opening 820 is an optical element shared with the light-emitting optical system 80, and is provided on the image side of the objective lens 821.
  • the fluorescence emitted from the objective lens 821 passes through the object-side opening 820.
  • the multi-fluorescence filter 810 is provided on the image side of the object-side opening 820.
  • the multi-fluorescence filter 810 is a so-called multi-bandpass filter that selectively passes light of multiple specific wavelengths.
  • the multi-fluorescence filter 810 passes light of the same wavelength as the fluorescence emitted by multiple types of fluorescent dyes contained in the sample, and blocks light of other wavelengths. In other words, the multi-fluorescence filter 810 selectively passes the fluorescence emitted by the fluorescent dyes contained in the sample.
  • the imaging lens 811 is provided on the image side of the multi-fluorescence filter 810.
  • the position and focal length of the imaging lens 811 are set so that the angle of incidence of the light entering the image-side opening 812, which is located on the image side of the imaging lens 811, is equal.
  • the light receiving element 813 which will be described later, can be placed on a common substrate 83 together with the light sources 801a and 801b of the light source section 801.
  • the image-side opening 812 is composed of a through hole provided in a plate-like member, and is provided on the image side of the imaging lens 811.
  • the image-side opening 812 adjusts the amount of light that passes through. Specifically, the image-side opening 812 passes a portion of the light that has passed through the imaging lens 811.
  • the light receiving element 813 is, for example, a photodiode, and is fixed to the substrate 83.
  • the light receiving element 813 converts the incident fluorescence into an electrical signal.
  • the light receiving optical system 81 is designed so that the light receiving element 813 can receive the fluorescence at a desired magnification.
  • the control unit 61 determines the amplification result of the nucleic acid based on the output of the light receiving element 813.
  • the driving unit 9 moves the fluorescence detection unit 8 under the control of the control unit 61.
  • the driving unit 9 has, for example, a motor 91 and a conversion mechanism 92 that converts the rotation of the motor 91 into linear motion.
  • the driving unit 9 moves the fluorescence detection unit 8 so that it faces one of the detection areas (areas including the low-temperature heater unit 642) of the thermal flow paths 3a to 3e in the cartridge C.
  • the driving unit 9 moves the fluorescence detection unit 8 to a position facing a detection region in a state where the nucleic acid amplification determination can be performed.
  • the order in which the driving unit 9 moves the fluorescence detection unit 8 may be predetermined or may be random.
  • the fluorescence detection process is performed when the fluorescence detection section 8 faces the detection area of the cartridge C.
  • the fluorescence detection process by the fluorescence detection device 7 is performed at a predetermined timing under the control of the control section 61.
  • the predetermined timing is, for example, immediately after the extension reaction step in the test.
  • the light source unit 801 (specifically, light sources 801a and 801b) emits a given light toward the detection area of the cartridge C under the control of the control unit 61.
  • Light emitted from the light source unit 801 (specifically, light sources 801a and 801b) enters the filter unit 802 (specifically, excitation filters 802a and 802b).
  • the light that passes through the filter section 802 passes through the object-side opening 820 and the objective lens 821, and is irradiated onto the detection area of the cartridge C.
  • the light irradiated to the detection area of the cartridge C is a parallel beam of light and is illuminated on the detection area of the cartridge C from any angle. Therefore, the detection area of the cartridge C is irradiated with little unevenness.
  • the fluorescent dye in cartridge C When the fluorescent dye in cartridge C is excited, the fluorescent dye emits fluorescence. Then, the light containing the fluorescence of the fluorescent dye is incident on objective lens 821.
  • the chief ray ⁇ of the light incident on objective lens 821 is parallel to the optical axis X of objective lens 821.
  • the light that passes through the objective lens 821 passes through the object-side opening 820 and enters the multi-fluorescence filter 810.
  • the fluorescence emitted by the fluorescent dye contained in the sample passes through the multi-fluorescence filter 810.
  • the light (fluorescence) that passes through the multi-fluorescence filter 810 passes through the imaging lens 811 and the image-side opening 812, and is imaged at a predetermined magnification on the light-receiving surface of the light-receiving element 813.
  • the light-receiving element 813 converts the received light (fluorescence) into an electrical signal and outputs it to the control unit 61.
  • the control unit 61 performs an amplification determination of the nucleic acid based on the electrical signal received from the light-receiving element 813.
  • the nucleic acid amplification test method includes an RNA extraction step, a reagent mixing step, and a thermal cycle step.
  • the thermal cycle step also includes an enzyme activation step, a reverse transcription step, a thermal denaturation step, an annealing step, and an extension reaction step.
  • the operator mixes the specimen collected from the patient with the sample in a container (not shown).
  • the specimen contains the target nucleic acid.
  • the sample in the container is then dripped from the sample drip port 20b of the cartridge C into the sample storage space 20a.
  • the sample drip port 20b and the air hole 20c are blocked by sealing members 51 and 52 (see Figure 1). Up to this point, this is a pre-processing step that is carried out outside the nucleic acid amplification device 6.
  • the operator sets the cartridge C containing the sample in the cartridge support part 62 of the nucleic acid amplification device 6.
  • the cartridge C is supported vertically by the cartridge support part 62 (see FIG. 8).
  • the cartridge C is supported by the cartridge support part 62 so that the low-temperature side heat part 33 is located on the lower side and the high-temperature side heat part 36 is located on the upper side.
  • Figure 13 is a time chart showing the temperature cycle in a nucleic acid amplification test.
  • the horizontal axis shows time
  • the vertical axis shows the temperature of the sample.
  • Step S1 in FIG. 13 is an RNA extraction step.
  • Step S2 in FIG. 13 is a reagent mixing step.
  • Step S3 in FIG. 13 is a thermal cycle step.
  • Step S31 in step S3 in FIG. 13 is an enzyme activation step.
  • Step S32 in step S3 in FIG. 13 is a reverse transcription step.
  • step S33 of step S3 in FIG. 13 the thermal denaturation step, annealing step, and extension reaction step are repeated a predetermined number of cycles.
  • the thermal denaturation step, annealing step, and extension reaction step are repeated a predetermined number of cycles.
  • the sample is contained in the sample storage space 20a, as shown by the diagonal grid in Figure 14A.
  • the control unit 61 controls the first low-temperature side valve drive unit 651 and the first high-temperature side valve drive unit 652 of the valve drive unit 65 to close the first low-temperature side valve 30 and the first high-temperature side valve 31.
  • the second valve 37 is in an open state.
  • the states of the first low temperature side valve 30, the first high temperature side valve 31, and the second valve 37 of the cartridge C and the position of the sample shown in Fig. 14A are as follows:
  • the state of the cartridge C shown in Fig. 14A is the state of the cartridge C when the RNA extraction step S1 is performed.
  • control unit 61 controls the low-temperature heater 642a to heat the low-temperature region R2 of the base unit 1 (specifically, the low-temperature side heat unit 33) to the first predetermined temperature T1.
  • the control unit 61 also controls the first high-temperature heater 641a to heat the first high-temperature region R11 of the base unit 1 (specifically, the high-temperature side heat unit 36 and the sample storage space 20a) to a second predetermined temperature T2.
  • control unit 61 may control the second high-temperature heater 641b to heat the second high-temperature region R12 (specifically, the preheat section 35) of the base section 1 to a third predetermined temperature T3. Note that the heating of the second high-temperature region R12 (preheat section 35) of the base section 1 by the preheat section 35 may be omitted.
  • the control unit 61 performs the RNA extraction step S1 in the time chart of FIG. 13.
  • the sample storage space 20a provided in the first high temperature area R11 is also heated to a second predetermined temperature T2 (e.g., 95°).
  • T2 e.g. 95°
  • the sample contained in the sample storage space 20a is also heated to the second predetermined temperature T2.
  • the envelope of the target nucleic acid is destroyed and the RNA of the target nucleic acid is extracted.
  • the heating time of the sample in the RNA extraction step S1 is, for example, 60 seconds.
  • the RNA extraction step S1 is a step that is performed when the target nucleic acid is RNA.
  • control unit 61 controls the first low-temperature side valve drive unit 651 and the first high-temperature side valve drive unit 652 of the valve drive unit 65 to open the first low-temperature side valve 30 and the first high-temperature side valve 31.
  • the control unit 61 also controls the second valve drive unit 653 to close the second valve 37.
  • first low temperature side valve 30 open state
  • First high-temperature side valve 31 open state
  • Second valve 37 closed state
  • Position of sample sample storage space 20a
  • a circulation flow path is formed in the cartridge C, connecting the sample storage section 20, the individual reagent support section 21, the first low-temperature side valve 30, the first parallel flow path 32a (specifically, the low-temperature side heat section 33, the pump 34, and the preheat section 35), and the first high-temperature side valve 31.
  • This circulation flow path corresponds to an example of a second circulation flow path.
  • the control unit 61 performs a first sample transfer process.
  • the first sample transfer process is a process for transferring the sample from the sample storage unit 20 to the individual reagent holding units 21.
  • the sample stored in the sample storage unit 20 is distributed to the multiple individual reagent holding units 21 by the pump 34.
  • the first sample transfer process is a process carried out between the RNA extraction process S1 and the reagent mixing process S2.
  • the control unit 61 drives the pump 34 to move the sample from the sample storage unit 20 to the individual reagent holding unit 21.
  • the direction in which the sample moves is opposite to the clockwise direction in FIG. 14A. Specifically, the sample moves from the sample storage unit 20 to the individual reagent holding unit 21 through the flow path element L1.
  • control unit 61 controls all pump drive units 661 corresponding to the thermal flow paths 3a to 3e to drive all pumps 34 corresponding to the thermal flow paths 3a to 3e.
  • the air and sample in the second circulation flow path move in conjunction with the movement of the air pushed out from the pump space 34c.
  • the sample in the sample storage section 20 moves toward the individual reagent carrier 21.
  • the sample is then sequentially stored in the individual reagent carrier 21 of each of the thermal flow paths 3a to 3e.
  • control unit 61 When the control unit 61 detects that the individual reagent carriers 21 of the thermal channels 3a to 3e are filled with sample, it stops driving the pump 34.
  • control unit 61 stops the pump drive unit 661 in the order of the thermal flow paths 3a to 3e in which the individual reagent holding units 21 are filled with sample.
  • a sample position detection unit (not shown) is provided to detect that a sample has been contained in the flow path between the individual reagent holding unit 21 and the first low-temperature side valve 30 in the flow path 2. Then, when this sample position detection unit detects a sample, the control unit 61 determines that the individual reagent holding unit 21 is filled with sample.
  • the configuration of the sample position detection unit may be the same as the configuration of the low-temperature side detection unit 671 of the sample position detection unit 67 described above.
  • the control unit 61 determines that the individual reagent holding unit 21 is filled with sample when the low temperature side detection unit 671 of the sample position detection unit 67 detects a sample. In the case of the second detection method, when the low temperature side detection unit 671 detects a sample, a portion of the sample is present downstream of the individual reagent holding unit 21. Therefore, the control unit 61 drives the pump 34 in the reverse direction to move a predetermined amount of sample upstream.
  • the sample in the first sample movement process, is not heated by the heater unit 64. In other words, in the first sample movement process, the sample is mainly affected by the temperature inside the device.
  • the time for the first sample movement process is approximately 10 to 30 seconds.
  • the cartridge C When the first sample transfer step is completed, the cartridge C is in the state shown in Fig. 14B.
  • the states of the first low temperature side valve 30, the first high temperature side valve 31, and the second valve 37 of the cartridge C and the position of the sample shown in Fig. 14B are as follows.
  • the state of the cartridge C shown in Fig. 14B is also the state of the cartridge C in the reagent mixing step S2 and the second sample transfer step performed after the first sample transfer step.
  • First low-temperature side valve 30 open state
  • First high-temperature side valve 31 open state
  • Second valve 37 closed state
  • Position of sample individual reagent support portion 21
  • the reagent mixing step S2 in the time chart of FIG. 13 is performed.
  • the control unit 61 drives the vibration unit 63.
  • the vibration unit 63 applies vibration to the individual reagent holder 21.
  • the temperature of the sample in the reagent mixing process S2 is, for example, a first predetermined temperature T1, which is the heating temperature of the low-temperature heater 642a.
  • the temperature of the sample in the reagent mixing process is not limited to the first predetermined temperature T1.
  • the second sample transfer process is a process for transferring the sample from the individual reagent support unit 21 to the low-temperature side heat unit 33.
  • the second sample transfer process is a process carried out between the reagent mixing process S2 and the thermal cycle process S3 (specifically, the enzyme activation process S31).
  • the control unit 61 drives the pump 34 to move the sample from the individual reagent holder 21 to the low-temperature side heat unit 33.
  • the direction in which the sample moves is opposite to the clockwise direction in FIG. 14B. Specifically, the sample moves from the individual reagent holder 21 to the low-temperature side heat unit 33 through the flow path element L3 (see FIG. 2).
  • control unit 61 controls all pump drive units 661 corresponding to the thermal flow paths 3a to 3e to drive all pumps 34 corresponding to the thermal flow paths 3a to 3e.
  • the sample is sequentially accommodated in the low-temperature side heat units 33 of each of the thermal flow paths 3a to 3e.
  • control unit 61 When the control unit 61 detects that a sample has been placed in the low-temperature side heat unit 33 of each of the thermal flow paths 3a to 3e, it stops driving the pump 34.
  • control unit 61 acquires information indicating that the sample has been accommodated in the low-temperature side heat unit 33 (low-temperature side accommodation information) based on the detection value of the low-temperature side detection unit 671 of the sample position detection unit 67.
  • control unit 61 determines whether or not a sample has been accommodated in the low-temperature side heat unit 33 based on the acquired low-temperature side accommodation information.
  • the control unit 61 stops the pump drive unit 661 in the order of thermal flow paths 3a to 3e in which a sample has been accommodated in the low-temperature side heat unit 33.
  • control unit 61 controls the first low-temperature side valve drive unit 651 and the first high-temperature side valve drive unit 652 to close the first low-temperature side valve 30 and the first high-temperature side valve 31.
  • the control unit 61 also controls the second valve drive unit 653 to open the second valve 37.
  • the circulation flow paths 32 of the thermal flow paths 3a to 3e in the flow path 2 become closed flow paths.
  • the circulation flow paths 32 in this state correspond to an example of the first circulation flow path.
  • the first circulation flow path is a flow path formed in the thermal cycle process S3 in the nucleic acid amplification test.
  • flow path 2 can alternatively be in a state where the first circulation flow path is formed (also referred to as the first state of flow path 2) or a state where the second circulation flow path is formed (also referred to as the second state of flow path 2), depending on the states of first low-temperature side valve 30, first high-temperature side valve 31, and second valve 37.
  • first low temperature side valve 30 closed state
  • First high-temperature side valve 31 closed state
  • Second valve 37 open state
  • Position of sample low-temperature side heat unit 33
  • the third sample transfer process is a process for transferring the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36.
  • the control unit 61 drives the pump 34 to move the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36.
  • the direction in which the sample moves is opposite to the clockwise direction in FIG. 14C.
  • the sample moves from the low-temperature side heat section 33 to the high-temperature side heat section 36 through flow path element L5 (see FIG. 2), pump 34, flow path element L6 (see FIG. 2), preheat section 35, and flow path element L7 (see FIG. 2).
  • control unit 61 controls all pump drive units 661 corresponding to the thermal flow paths 3a to 3e to drive all pumps 34 corresponding to the thermal flow paths 3a to 3e.
  • the sample is sequentially accommodated in the high-temperature side heat units 36 of each of the thermal flow paths 3a to 3e.
  • control unit 61 When the control unit 61 detects that a sample has been placed in the high-temperature side heat unit 36 of each of the thermal flow paths 3a to 3e, it stops driving the pump 34.
  • control unit 61 acquires information indicating that the sample has been accommodated in the high-temperature side heat unit 36 (high-temperature side accommodation information) based on the detection value of the high-temperature side detection unit 672 of the sample position detection unit 67.
  • control unit 61 determines whether or not a sample has been accommodated in the high-temperature side heat unit 36 based on the acquired high-temperature side accommodation information.
  • the control unit 61 stops the pump drive unit 661 in the order of thermal flow paths 3a to 3e in which a sample has been accommodated in the high-temperature side heat unit 36.
  • the cartridge C When the third sample movement step is completed, the cartridge C is in the state shown in Fig. 14D.
  • the states of the first low temperature side valve 30, the first high temperature side valve 31, and the second valve 37 of the cartridge C shown in Fig. 14D, and the position of the sample are as follows.
  • First low-temperature side valve 30 closed state
  • First high-temperature side valve 31 closed state
  • Second valve 37 open state
  • Position of sample high-temperature side heat unit 36
  • the enzyme activation step S31 is a step for increasing the activity of the enzyme in the sample.
  • the control unit 61 heats the sample in the high-temperature side heat unit 36 to a second predetermined temperature T2 (e.g., 95 degrees) by the first high-temperature heater 641a for a predetermined time (e.g., 60 seconds).
  • the control section 61 keeps the sample at the high-temperature side heat section 36 for a predetermined time.
  • the fourth sample transfer process is a process for transferring the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33.
  • the fourth sample transfer process is a process carried out between the enzyme activation process S31 and the reverse transcription process S32.
  • the control unit 61 drives the pump 34 to move the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33.
  • the direction in which the sample moves is opposite to the clockwise direction in FIG. 14D. Specifically, the sample moves from the high-temperature side heat unit 36 to the low-temperature side heat unit 33 through the flow path element L8 (see FIG. 2).
  • control unit 61 controls all pump drive units 661 corresponding to the thermal flow paths 3a to 3e to drive all pumps 34 corresponding to the thermal flow paths 3a to 3e.
  • the sample is sequentially accommodated in the low-temperature side heat units 33 of each of the thermal flow paths 3a to 3e.
  • control unit 61 When the control unit 61 detects that a sample has been placed in the low-temperature side heat unit 33 of each of the thermal flow paths 3a to 3e, it stops driving the pump 34.
  • control unit 61 acquires information indicating that the sample has been accommodated in the low-temperature side heat unit 33 (low-temperature side accommodation information) based on the detection value of the low-temperature side detection unit 671 of the sample position detection unit 67.
  • control unit 61 determines whether or not a sample has been accommodated in the low-temperature side heat unit 33 based on the acquired low-temperature side accommodation information.
  • the control unit 61 stops the pump drive unit 661 in the order of thermal flow paths 3a to 3e in which a sample has been accommodated in the low-temperature side heat unit 33.
  • first low temperature side valve 30 closed state
  • First high-temperature side valve 31 closed state
  • Second valve 37 open state
  • Position of sample low-temperature side heat unit 33
  • the control unit 61 performs the reverse transcription step S32 in the time chart of FIG. 13 in the state shown in FIG. 14C.
  • the reverse transcription step S32 is a step for starting the so-called reverse transcription reaction between the RNA in the sample and an enzyme (reverse transcriptase).
  • control unit 61 heats the sample in the low-temperature side heating unit 33 to a first predetermined temperature T1 (e.g., 60 degrees) using the low-temperature heater 642a for a predetermined time (e.g., 60 seconds).
  • a first predetermined temperature T1 e.g. 60 degrees
  • a predetermined time e.g. 60 seconds
  • the control section 61 keeps the sample in the low-temperature side heat section 33 for a specified time.
  • control unit 61 drives the pump 34 to move the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36, as in the third sample movement process.
  • the sample passes through the preheat unit 35, it is heated by the preheat unit 35 and the temperature of the sample increases.
  • the heating temperature of the second high-temperature heater 641b that heats the preheat section 35 is the third predetermined temperature T3, so when the sample passes through the preheat section 35, the temperature of the sample rises to a maximum of the third predetermined temperature T3.
  • first low-temperature side valve 30 closed state
  • First high-temperature side valve 31 closed state
  • Second valve 37 open state
  • Position of sample high-temperature side heat unit 36
  • the control unit 61 performs the thermal denaturation step at step S33 in the time chart of FIG. 13.
  • the control unit 61 heats the sample in the high-temperature side heat unit 36 to a second predetermined temperature T2 (e.g., 95°C) by the first high-temperature heater 641a for a predetermined time (e.g., 2 seconds).
  • the control section 61 keeps the sample at the high-temperature side heat section 36 for a predetermined time.
  • the sample is heated to the third predetermined temperature T3 in the preheat section 35, so it is already at a high temperature when it flows into the high-temperature side heat section 36. This makes it possible to shorten the time it takes to heat the sample in the high-temperature side heat section 36.
  • control unit 61 drives the pump 34 to move the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33, similar to the fourth sample movement process.
  • first low-temperature side valve 30 closed state
  • First high-temperature side valve 31 closed state
  • Second valve 37 open state
  • Position of sample low-temperature side heat unit 33
  • the control unit 61 performs an annealing step and an extension reaction step at step S33 in the time chart of FIG. 13.
  • the control unit 61 heats the sample in the low-temperature side heat unit 33 to a first predetermined temperature T1 (e.g., 60 degrees) by the low-temperature heater 642a for a predetermined time (e.g., 5 seconds).
  • the control section 61 keeps the sample in the low-temperature side heat section 33 for a specified time.
  • control unit 61 performs an amplification determination process based on the detection value of the fluorescence detection device 7 at a predetermined timing, along with the annealing process and the extension reaction process.
  • the amplification determination process is as described above.
  • the predetermined timing is, for example, immediately after the extension reaction process.
  • the thermal denaturation step, annealing step, and extension reaction step are repeatedly performed for a predetermined number of cycles (e.g., 40 cycles).
  • the control unit 61 performs an amplification determination process along with the annealing step and extension reaction step in each cycle.
  • the control unit 61 counts the number of cycles of the thermal cycle process, for example, based on the number of detections by the low-temperature side detection unit 671. Alternatively, the control unit 61 may count the number of cycles of the thermal cycle process, for example, based on the number of detections by the fluorescence detection device 7.
  • one test is completed in approximately 900 seconds (15 minutes).
  • the sample circulates in a predetermined direction through the circulation flow path 32 of the cartridge C. This eliminates the need for cumbersome controls such as switching the sample movement direction, and shortens the time required for testing.
  • the sample moves cyclically in a predetermined direction through the circulation flow path 32 of the cartridge C. Therefore, it is relatively easy to detect the position of the sample, control the position of the sample, and control the drive of the nucleic acid amplifier 6.
  • the configuration of the nucleic acid amplifier 6 can be made relatively simple. Therefore, the manufacturing cost of the nucleic acid amplifier 6 can be reduced.
  • the cartridge C of this embodiment has an individual reagent carrying section 21 that carries in advance an individual reagent corresponding to the target nucleic acid. This makes it possible to omit the pre-treatment step of mixing the sample and the individual reagent outside the cartridge, which has been conventionally performed. As a result, it is possible to reduce the testing time while reducing the burden on the operator.
  • the flow path 2 of the cartridge C is completely blocked from the outside air by the sealing members 50, 51, and 52. This ensures that the test can be performed reliably while reliably preventing the leakage of viruses during testing.
  • the cartridge C of this embodiment has multiple thermal flow paths 3a to 3e. This allows multiple items to be tested in one test. As a result, it is possible to improve the efficiency of testing while reducing the workload of the operator and the burden on the subject.
  • the nucleic acid amplification device 6 of this embodiment supports the cartridge C vertically so that the low-temperature side heat section 33 is positioned at the bottom. This makes it easier to collect the sample at the low-temperature side heat section 33, which is the starting position for the thermal cycle process. In addition, the effect of gravity acting on the sample makes it easier to maintain the sample in the flow channel 2 as a single mass.
  • the low-temperature side heat section 33 is positioned lower than the high-temperature side heat section 36, the low-temperature side heat section 33 is less susceptible to the heat of the high-temperature heater section 641 during testing. As a result, stable test results can be obtained.
  • the sample circulates through the circulation flow path 32 in a path that passes through the pump 34.
  • the sample may circulate through the circulation flow path 32 in a path that does not pass through the pump 34.
  • the sample may circulate through the circulation flow path 32 in a path that does not pass through the pump 34 during the thermal cycle processing in the nucleic acid amplification testing.
  • Variant 1 of the nucleic acid amplification testing method will be described.
  • the drawings used in the above-mentioned embodiment 1 will be appropriately cited.
  • the configuration of the cartridge C used in the modified example 1 of the nucleic acid amplification testing method is the same as the configuration of the cartridge C in the above-mentioned embodiment 1.
  • the configuration of the cartridge may be different from the configuration of the cartridge C in embodiment 1.
  • the configuration of the nucleic acid amplifier 6 used in the modified example 1 of the nucleic acid amplification testing method is also substantially the same as the configuration of the nucleic acid amplifier 6 in the above-mentioned embodiment 1.
  • the configuration of the nucleic acid amplifier may be different from the configuration of the nucleic acid amplifier 6 in embodiment 1.
  • the operation of the nucleic acid amplification device 6 that performs the nucleic acid amplification testing method differs from the operation of the nucleic acid amplification device 6 in the above-mentioned embodiment 1.
  • modified example 1 of the nucleic acid amplification testing method will be described, focusing on the configuration that differs from the nucleic acid amplification testing method according to the above-mentioned embodiment 1.
  • nucleic acid amplification testing method variant 1 the explanation of the configuration that is the same as that of the nucleic acid amplification testing method of embodiment 1 described above may be appropriately cited from the explanation of the nucleic acid amplification testing method of embodiment 1.
  • the sample repeatedly moves back and forth between the first position and the second position in the circulation flow path 32.
  • the pump 34 of the cartridge C repeatedly moves the sample back and forth between the first position and the second position in the circulation flow path 32.
  • the control unit 61 of the nucleic acid amplification device 6 drives the pump 34 to repeatedly move the sample back and forth between the first position and the second position in the circulation flow path 32.
  • the repeated movement between the first position and the second position in the circulation flow channel 32 is also included in the concept of the sample circulating.
  • the concept of the sample circulating is not limited to the sample moving circularly in one direction in the circulation flow channel 32, but also includes the sample moving back and forth in the circulation flow channel 32.
  • the processing of each step of the nucleic acid amplification testing method performed in the nucleic acid amplifier 6 is controlled by the control unit 61 (see FIG. 8).
  • control unit 61 also performs the RNA extraction step S1 and the reagent mixing step S2 in the time chart of FIG. 13, as in the nucleic acid amplification testing method of embodiment 1.
  • the explanation of the RNA extraction step S1 and the reagent mixing step S2 in embodiment 1 may be appropriately used for the explanation of the RNA extraction step S1 and the reagent mixing step S2 in this modified example.
  • control unit 61 performs the thermal cycle step S3 in the time chart of FIG. 13 after the reagent mixing step S2.
  • the thermal cycle step S3 in this modified example is described below.
  • the second sample transfer process is a process for transferring the sample from the individual reagent holder 21 to the low-temperature side heat unit 33.
  • the explanation of the second sample transfer process may be appropriately applied to the explanation of the second sample transfer process in the first embodiment.
  • the cartridge C is in the state shown in FIG. 14C.
  • the third sample transfer process is a process for transferring the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36.
  • the sample travels from the low-temperature side heat unit 33 through the second parallel flow path 32b toward the high-temperature side heat unit 36. In other words, the sample does not pass through the pump 34.
  • the control unit 61 drives the pump 34 to move the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36.
  • the movement direction of the sample in the circulation flow path 32 is the clockwise direction in FIG. 14C (the direction indicated by the arrow Y8 in FIG. 14C).
  • the driving direction of pump 34 in the third sample movement process is also referred to as the first driving direction of pump 34.
  • control unit 61 performs the enzyme activation step S31 in the time chart of FIG. 13 in the state shown in FIG. 14D.
  • the explanation of the enzyme activation step S31 may be appropriately cited from the explanation of the enzyme activation step S31 in the first embodiment.
  • the fourth sample transfer process is a process for transferring the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33.
  • the sample travels from the high-temperature side heat unit 36 through the second parallel flow path 32b toward the low-temperature side heat unit 33. In other words, the sample does not pass through the pump 34.
  • the control unit 61 drives the pump 34 to move the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33.
  • the movement direction of the sample in the circulation flow path 32 is the opposite direction to the clockwise direction in FIG. 14C (the direction indicated by the arrow Y1 in FIG. 14C).
  • the driving direction of pump 34 in the fourth sample movement process is also referred to as the second driving direction of pump 34.
  • the second driving direction of pump 34 is opposite to the first driving direction of pump 34.
  • the direction of sample movement in the third sample movement process (in other words, the driving direction of pump 34) is opposite to the direction of sample movement in the fourth sample movement process (in other words, the driving direction of pump 34).
  • control unit 61 also performs the reverse transcription step S32 in the time chart of FIG. 13 in the state shown in FIG. 14C.
  • the description of the reverse transcription step S32 in the first embodiment may be used as appropriate for the description of the reverse transcription step S32.
  • control unit 61 drives the pump 34 in the first driving direction to move the sample from the low-temperature side heat unit 33 to the high-temperature side heat unit 36, similar to the third sample movement process.
  • the sample travels from the low-temperature side heat section 33 through the second parallel flow path 32b toward the high-temperature side heat section 36. In other words, the sample does not pass through the pump 34. In the case of this modified example, the sample passes through the high-temperature side heat section 36 and moves to the preheat section 35.
  • the control section 61 drives the pump 34 in the direction opposite to the first driving direction (i.e., the second driving direction) to move the sample from the preheat section 35 to the high-temperature side heating section 36.
  • the sample moves from the low-temperature side heat section 33 to the preheat section 35.
  • the sample is then heated in the preheat section 35.
  • the sample then moves from the preheat section 35 to the high-temperature side heat section 36. Therefore, the state (specifically, the temperature) of the sample accommodated in the high-temperature side heat section 36 is substantially the same as the state (specifically, the temperature) of the sample accommodated in the high-temperature side heat section 36 in the above-described first embodiment.
  • the preheat section 35 may be provided in the second parallel flow path 32b.
  • the control section 61 can drive the pump 34 only in the first driving direction to move the sample from the low-temperature side heat section 33 to the high-temperature side heat section 36.
  • control unit 61 performs a thermal denaturation process at step S33 in the time chart of FIG. 13.
  • the description of the thermal denaturation process may be appropriately referenced from the description of the thermal denaturation process in embodiment 1.
  • control unit 61 drives the pump 34 in the second driving direction to move the sample from the high-temperature side heat unit 36 to the low-temperature side heat unit 33, similar to the fourth sample movement process.
  • the sample moves from the high-temperature side heat section 36 through the second parallel flow path 32b to the low-temperature side heat section 33. In other words, the sample does not pass through the pump 34.
  • the cartridge C is in the state shown in Figure 14C.
  • control unit 61 performs an annealing step and an extension reaction step in step S33 in the time chart of FIG. 13.
  • the explanation of the annealing step and the extension reaction step may be appropriately cited from the explanation of the annealing step and the extension reaction step in embodiment 1.
  • control unit 61 also performs an amplification determination process based on the detection value of the fluorescence detection device 7 at a predetermined timing in addition to the annealing process and the extension reaction process.
  • the amplification determination process is as described above.
  • the predetermined timing is, for example, immediately after the extension reaction process.
  • the thermal denaturation step, annealing step, and extension reaction step are repeatedly performed for a predetermined number of cycles (e.g., 40 cycles).
  • the control unit 61 performs an amplification determination process along with the annealing step and extension reaction step in each cycle.
  • the sample does not pass through the pump 34 during thermal cycling.
  • thermal cycling can be performed without the sample passing directly below the pump section 34.
  • the sample does not pass directly below the pump section 34, so the sample is not directly compressed by the pump section 34. This makes it possible to prevent air bubbles from being generated in the sample. As a result, highly accurate detection is possible. Furthermore, in this modified example, the sample also passes through the preheat section 35. This makes it possible to shorten the inspection time.
  • Fig. 15 is a schematic cross-sectional view of a nucleic acid amplifier 6A according to embodiment 2 of the present invention.
  • the nucleic acid amplifier 6A differs from the nucleic acid amplifier 6 shown in Fig. 8 in the manner in which the cartridge C is supported. Due to this difference, a portion of the configuration of the nucleic acid amplifier 6A differs from the configuration of the nucleic acid amplifier 6 shown in Fig. 8.
  • the configuration of the nucleic acid amplifier 6A will be described below, focusing on the parts that differ from the nucleic acid amplifier 6 shown in Fig. 8.
  • the nucleic acid amplification device 6A has a housing 60A, a control unit 61A, a cartridge support unit 62A, a vibration unit 63A, a heater unit 64A, a valve drive unit 65A, a liquid delivery unit 66A, a sample position detection unit 67A, and a fluorescence detection device 7A.
  • the housing 60A is box-shaped and has a storage space 601A.
  • the storage space 601A contains the elements 61A to 67A and 7A that make up the nucleic acid amplification device 6A.
  • the control unit 61A has a first control unit 611A and a second control unit 612A.
  • the configurations of the first control unit 611A and the second control unit 612A are similar to the configurations of the first control unit 611 and the second control unit 612 of the nucleic acid amplification device 6 shown in FIG. 8.
  • the cartridge support portion 62A is supported by the housing 60A and is a member that supports the cartridge C.
  • the configuration of the cartridge support portion 62A is substantially the same as the configuration of the cartridge support portion 62 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the cartridge support portion 62A supports the cartridge C horizontally.
  • the first and second main surfaces of the cartridge C face vertically.
  • the vibration unit 63A is supported by the housing 60A.
  • the configuration of the vibration unit 63A is almost the same as the configuration of the vibration unit 63 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the vibration unit 63A is provided at a position facing the individual reagent holding unit 21 of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the heater unit 64A is supported by the housing 60A.
  • the configuration of the heater unit 64A is substantially the same as the configuration of the heater unit 64 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the low-temperature heater 642b is provided at a position facing the low-temperature region R2 (specifically, the low-temperature side heat section 33) of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the first high temperature heater 641c is supported at a position facing the first high temperature region R11 (specifically, the high temperature side heat section 36) of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the second high-temperature heater 641d is supported at a position facing the second high-temperature region R12 (specifically, the preheat section 35) of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the valve drive unit 65A is supported by the housing 60A.
  • the configuration of the valve drive unit 65A is substantially the same as the configuration of the valve drive unit 65 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the pressing portion 651b of the first low-temperature side valve driving portion 651A is supported at a position facing the pressed portion 30c of the first low-temperature side valve 30 in the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the pressing portion 652b of the first high temperature side valve driving portion 652A is provided at a position facing the pressed portion 31c of the first high temperature side valve 31 in the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the pressing portion 653b of the second valve driving portion 653A is provided at a position facing the pressed portion 37c of the second valve 37 in the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the liquid delivery unit 66A is supported by the housing 60A.
  • the configuration of the liquid delivery unit 66A is substantially the same as the configuration of the liquid delivery unit 66 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the pump drive unit 661A of the liquid delivery unit 66A is provided at a position facing the pump 34 in a predetermined direction (the vertical direction in this embodiment).
  • the sample position detection unit 67A is supported by the housing 60A.
  • the configuration of the sample position detection unit 67 is substantially the same as the configuration of the sample position detection unit 67 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the low-temperature side detection section 671A of the sample position detection section 67A is provided at a position facing the low-temperature side heating section 33 of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the high temperature side detection section 672A of the sample position detection section 67A is provided at a position facing the high temperature side heating section 36 of the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the fluorescence detection device 7A is supported by the housing 60A.
  • the configuration of the fluorescence detection device 7A is almost the same as the configuration of the fluorescence detection device 7 in the nucleic acid amplification device 6 shown in FIG. 8.
  • the fluorescence detection unit 8 of the fluorescence detection device 7A is provided at a position facing the detection area (low-temperature side heat unit 33) in the cartridge C in a predetermined direction (vertical direction in this embodiment) during testing.
  • the nucleic acid amplifier 6A of this embodiment which has the above-described configuration, can also shorten the time required for testing.
  • the rest of the configuration and the actions and effects of the nucleic acid amplifier 6A are the same as those of the nucleic acid amplifier 6 of embodiment 1.
  • a nucleic acid amplifier 6B according to embodiment 3 will be described with reference to Figures 16 to 18.
  • the configuration of a pump driver 661B is different from the pump driver 661 of the nucleic acid amplifier 6 according to embodiment 1.
  • the configuration other than the pump driver 661B is the same as that of the nucleic acid amplifier 6 according to embodiment 1.
  • Thermal flow path 3g is shown in Figures 16 to 18.
  • the circulation flow path 32A of thermal flow path 3g has a pair of pumps 34C, 34D.
  • the configuration of thermal flow path 3g is slightly different from the configuration of thermal flow path 3a in embodiment 1.
  • the configurations of the flow paths other than thermal flow path 3g are almost the same as the configuration of flow path 2 in embodiment 1.
  • Pumps 34C and 34D are arranged in parallel in circulation flow path 32A.
  • the configuration of pumps 34C and 34D is substantially the same as the configuration of pump 34A of modification 1 shown in Figures 5 and 6.
  • pumps 34C and 34D each have curved upper pump space forming members 34k and 34m. Therefore, pump spaces 34n and 34p defined by upper pump space forming members 34k and 34m are also curved.
  • the upper pump space forming members 34k, 34m are each arranged along an arc centered on a common center point.
  • the pump drive unit 661B has a base 662a and multiple (in this embodiment, three) roller members 662b.
  • the base 662a is a part that is fixed to the housing 60 (see FIG. 8).
  • the base 662a is also rotatable about a central axis A1.
  • the central axis A1 is perpendicular to the main surface of the cartridge C1 during inspection.
  • the base 662a is also a member that rotatably supports the roller member 662b.
  • roller members 662b are arranged at equal intervals (120 degree intervals in this embodiment) in the rotation direction of the base 662a.
  • Each roller member 662b has a shaft portion 662c and a pressure roller 662d.
  • the shaft portion 662c and the pressure roller 662d are integrally formed.
  • the shaft portion 662c is supported by the base portion 662a in a state in which it can rotate around the central axis A2 of the shaft portion 662c.
  • the pressure roller 662d is frustum-shaped and is integrally formed at the tip of the shaft portion 662c.
  • the pressure roller 662d has a pressure surface 662e on its outer circumferential surface.
  • the pressure surface 662e is the part that presses the upper pump space forming members 34k, 34m of the pumps 34C, 34D when the pumps 34C, 34D are driven.
  • the pump drive unit 661A is arranged to cover the upper pump space forming members 34k, 34m of the pumps 34C, 34D from a predetermined direction. In this state, the pressing surface 662e of the pressing roller 662d can abut against the upper pump space forming members 34k, 34m of the pumps 34C, 34D.
  • the pump drive unit 661A and the cartridge C1 are positioned farther apart than they were during testing.
  • Figure 18 is a schematic diagram showing the contact positions D1 and D2 between the pressure roller 662d of the pump drive unit 661A and the upper pump space forming members 34k and 34m of the pumps 34C and 34D.
  • One of the three pressure rollers 662d (the upper pressure roller 662d in FIG. 18) abuts against the upper pump space forming member 34k of the pump 34C at abutment position D1.
  • the upper pressure roller 662d in FIG. 18 presses against the upper pump space forming member 34k of the pump 34C at abutment position D1.
  • one of the three pressure rollers 662d (the lower left pressure roller 662d in FIG. 18) abuts against the upper pump space forming member 34m of the pump 34D at abutment position D2.
  • the lower left pressure roller 662d in FIG. 18 presses against the upper pump space forming member 34m of the pump 34D at abutment position D2.
  • the three pressure rollers 662d each move in the direction of the arrow Y3 while rolling.
  • the contact positions D1 and D2 also move in the direction of the arrow Y3.
  • the upper pump space forming member 34k of the pump 34C and the upper pump space forming member 34m of the pump 34D are crushed by the pressure rollers 662d.
  • the air and/or sample in the pump spaces 34n and 34p of the pumps 34C and 34D move in the sample circulation direction (the direction indicated by the arrow Y7 in FIG. 16).
  • the air and sample in the thermal flow path 3g move in the circulation direction in conjunction with the movement of the air and/or sample in the pump spaces 34n and 34p.
  • the three pressure rollers 662d sequentially drive pumps 34C and 34D in response to the rotation of base 662a.
  • the direction of the frictional force that the three pressure rollers 662d apply to pumps 34C and 34D is along the extension direction of pumps 34C and 34D.
  • the pump drive unit 661A can sequentially drive pumps 34C and 34D using the three pressure rollers 662d. This allows the sample to move efficiently through the flow path.
  • the pump drive unit 661A of this embodiment can be applied to a cartridge having a curved pump.
  • the number of pumps is not limited to two, and may be one. Other configurations and actions/effects are substantially the same as those of the first embodiment.
  • the present invention contributes to improving the efficiency of testing in nucleic acid amplification devices such as real-time PCR devices, and has great industrial applicability.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Sustainable Development (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Récipient de test utilisé pour un test d'amplification d'acide nucléique réalisé dans un dispositif d'amplification d'acide nucléique. Le récipient de test comprend : un substrat ; et une voie d'écoulement pourvue d'un substrat et dans laquelle est logé un échantillon contenant de l'acide nucléique. La voie d'écoulement comprend : une voie d'écoulement thermique incluant une voie d'écoulement de circulation dans laquelle l'échantillon circule ; et une pompe présente dans la voie d'écoulement de circulation et qui déplace l'échantillon.
PCT/JP2023/032960 2022-09-28 2023-09-11 Récipient de test, dispositif d'amplification d'acide nucléique et procédé de test d'amplification d'acide nucléique WO2024070614A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2022154913 2022-09-28
JP2022-154913 2022-09-28

Publications (1)

Publication Number Publication Date
WO2024070614A1 true WO2024070614A1 (fr) 2024-04-04

Family

ID=90477507

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/032960 WO2024070614A1 (fr) 2022-09-28 2023-09-11 Récipient de test, dispositif d'amplification d'acide nucléique et procédé de test d'amplification d'acide nucléique

Country Status (1)

Country Link
WO (1) WO2024070614A1 (fr)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0775544A (ja) * 1993-07-28 1995-03-20 Perkin Elmer Corp:The 核酸増幅反応を実施する装置、化学連鎖反応を実施する装置、変性、アニーリングおよびエクステンションプロセスを包含する核酸増幅反応を同時に実施する装置、ならびに核酸増幅反応を実施する方法
US20080176292A1 (en) * 2007-01-23 2008-07-24 Texas A&M University System Portable buoyancy driven pcr thermocycler
US20100267127A1 (en) * 2009-04-16 2010-10-21 Electronics And Telecommunications Research Institute Polymerase chain reaction apparatus
KR20130008266A (ko) * 2011-07-12 2013-01-22 강원대학교산학협력단 중력유동방식 ｐｃｒ 증폭 장치
JP2021073983A (ja) * 2019-09-09 2021-05-20 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ スマートフォンｐｃｒデバイス
US20210388428A1 (en) * 2020-06-12 2021-12-16 Electronics And Telecommunications Research Institute Method and device for amplifying and detecting gene

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0775544A (ja) * 1993-07-28 1995-03-20 Perkin Elmer Corp:The 核酸増幅反応を実施する装置、化学連鎖反応を実施する装置、変性、アニーリングおよびエクステンションプロセスを包含する核酸増幅反応を同時に実施する装置、ならびに核酸増幅反応を実施する方法
US20080176292A1 (en) * 2007-01-23 2008-07-24 Texas A&M University System Portable buoyancy driven pcr thermocycler
US20100267127A1 (en) * 2009-04-16 2010-10-21 Electronics And Telecommunications Research Institute Polymerase chain reaction apparatus
KR20130008266A (ko) * 2011-07-12 2013-01-22 강원대학교산학협력단 중력유동방식 ｐｃｒ 증폭 장치
JP2021073983A (ja) * 2019-09-09 2021-05-20 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ スマートフォンｐｃｒデバイス
US20210388428A1 (en) * 2020-06-12 2021-12-16 Electronics And Telecommunications Research Institute Method and device for amplifying and detecting gene

Similar Documents

Publication Publication Date Title
AU2010237532B2 (en) Optical detection system for monitoring rtPCR reaction
US20190004054A1 (en) Assays
EP2032722B1 (fr) Systèmes de détection et de préparation d'échantillons sans pcr à des fins d'identification et d'analyse biologique de vitesse élevée
US11198122B2 (en) Diagnostic test assembly, apparatus, method
CN112638241A (zh) 便携式诊断设备及其方法
OA11191A (en) Laboratory in a disk
CA2521999A1 (fr) Appareil et procede de dosage utilisant des reseaux microfluidiques
JPWO2005116202A1 (ja) 反応容器、反応測定装置、および液回転処理装置
JPWO2006013926A1 (ja) 反応容器、反応容器液導入装置、液導入反応測定装置、および液導入装置
JP5567526B2 (ja) 分析装置および分析方法
US20100227384A1 (en) Portable Genomic Analyzer
CN111778156A (zh) 高通量pcr微滴荧光检测装置及方法
WO2024070614A1 (fr) Récipient de test, dispositif d'amplification d'acide nucléique et procédé de test d'amplification d'acide nucléique
KR102102773B1 (ko) 초소형 광학요소를 적용한 현장진단용 형광검출장치
EP2225548B1 (fr) Système et procédé de détection
TW201237410A (en) Apparatus and method for detecting biochemical reaction

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23871850

Country of ref document: EP

Kind code of ref document: A1