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WO2020174918A1 - Dispositif d'amplification d'acide nucléique - Google Patents

Dispositif d'amplification d'acide nucléique Download PDF

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Publication number
WO2020174918A1
WO2020174918A1 PCT/JP2020/001355 JP2020001355W WO2020174918A1 WO 2020174918 A1 WO2020174918 A1 WO 2020174918A1 JP 2020001355 W JP2020001355 W JP 2020001355W WO 2020174918 A1 WO2020174918 A1 WO 2020174918A1
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WIPO (PCT)
Prior art keywords
temperature
sample
control unit
nucleic acid
temperature control
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2020/001355
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English (en)
Japanese (ja)
Inventor
玉置　裕一
満田　綾子
理太郎 小熊
章人 澤井
吉田　正樹
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PHC Holdings Corp
Original Assignee
PHC Holdings Corp
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Filing date
Publication date
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Publication of WO2020174918A1 publication Critical patent/WO2020174918A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • 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
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control

Definitions

  • the present invention relates to a nucleic acid amplification device.
  • PCR Polymerase Chain Reaction
  • a nucleic acid such as DNA (Doxyribonucleic Acid: deoxyribonucleic acid)
  • a nucleic acid amplification device for amplification is known.
  • a nucleic acid amplification device that includes a nucleic acid detection unit in addition to a nucleic acid amplification mechanism and can detect the amplified nucleic acid in real time (see, for example, Patent Document 1).
  • a nucleic acid amplification device is called a real-time PCR device.
  • a real-time PCR device controls the temperature of a reaction sample containing DNA, a fluorescent substance, etc. to amplify DNA.
  • the real-time PC device irradiates with excitation light that excites the reaction sample, and quantitatively measures the amplified DNA based on the fluorescence generated from the reaction sample.
  • the real-time PCR apparatus is widely used not only in the research field but also in various inspection fields.
  • Patent Document 1 Japanese Patent Laid-Open No. 201 0 _ 81 898
  • nucleic acid amplification is performed by repeating a cycle including a heat denaturation step, an annealing step, and an extension reaction step a plurality of times. Therefore, it takes a relatively long time (for example, 40 minutes) to react one sample. Therefore, if the number of samples is large, it will take a long time. ⁇ 02020/174918 2 ⁇ (: 170?2020/001355
  • the present invention has been made in view of such circumstances, and an object of the present invention is to provide a nucleic acid amplification device capable of efficiently reacting a plurality of samples in a short time.
  • a nucleic acid amplification apparatus comprises: a first temperature control unit for heating and cooling a first sample containing nucleic acid; and a second temperature control unit for heating and cooling a second sample containing nucleic acid. And a control that controls the first temperature control unit and the second temperature control unit, and starts the control of the second temperature control unit during the control of the first temperature control unit. And a section.
  • nucleic acid amplifier capable of efficiently reacting a plurality of samples in a short time.
  • FIG. 1 A schematic vertical cross-sectional view of a nucleic acid amplification device according to an embodiment.
  • FIG. 2 A schematic cross-sectional view of a nucleic acid amplification device according to an embodiment.
  • FIG. 4 A plan view of the area around the container.
  • FIG. 58 A plan view of an example of the container.
  • FIG. 58 A side view of an example of the container.
  • FIG. 68 A plan view of an example of the container.
  • FIG. 68 A side view of an example of the container.
  • FIG. 7 Time chart showing the temperature cycle in the polymerase chain reaction.
  • FIG. 8 An evening chart showing a temperature cycle of control executed by the nucleic acid amplification device according to the embodiment.
  • FIG. 90 Display example on the operation display unit included in the nucleic acid amplification device according to the embodiment.
  • FIG. 9 A display example on the operation display unit included in the nucleic acid amplification device according to the embodiment.
  • FIG. 9D _ Display example on the operation display unit included in the nucleic acid amplification device according to the embodiment.
  • FIG. 90] _ Display example on the operation display unit included in the nucleic acid amplification device according to the embodiment.
  • FIG. 10 A front view of the vicinity of a portion where a container is placed in a nucleic acid amplification device according to another embodiment.
  • FIG. 1 is a vertical cross-sectional view of a nucleic acid amplification device 1 according to an embodiment.
  • 2 is a cross-sectional view of the nucleic acid amplification device 1 according to the embodiment.
  • the nucleic acid amplification device 1 has an opening, a housing 11 for receiving a sample, and a cover 12 for opening and closing the opening of the housing 11. Covers 1 and 2 are lockable and cannot be opened when locked.
  • an optical measuring device 13 Inside the housing 11, an optical measuring device 13, a reflecting mirror 14 and a Fresnel lens 15 are arranged.
  • the optical measuring device 13 includes a lamp that emits excitation light that is irradiated onto the sample, and a plurality of two-dimensionally arranged photodetection elements that receive the fluorescence emitted from the sample (for example, 0000 or 001 ⁇ /1 ⁇ 3) etc.
  • the reflecting mirror 14 reflects the excitation light emitted from the optical measurement device 13 toward the Fresnel lens 15 and reflects the fluorescence emitted from the sample toward the optical measurement device 13.
  • the Fresnel lens 15 corrects the excitation light reflected by the reflecting mirror 14 so that the plurality of samples are uniformly irradiated with the excitation light. Further, the Fresnel lens 15 corrects the optical path of the fluorescence so that the fluorescence emitted from the sample can be uniformly measured by the optical measuring device 13.
  • An operation display unit 16 including a touch panel or the like is attached to the outside of the housing 11.
  • the operation display unit 16 receives an input operation to the nucleic acid amplification device 1 and displays the state of the nucleic acid amplification device 1.
  • the operation display unit 16 includes an operation unit including buttons for accepting operations and a liquid crystal panel for displaying. ⁇ 02020/174918 4 ⁇ (: 170?2020/001355
  • a control unit 17 that integrally controls the nucleic acid amplification device 1 is arranged inside the housing 11.
  • the nucleic acid amplification device 1 may be connected to an external computer by wire or wirelessly and controlled by this computer instead of being provided with the control unit 17.
  • a first mounting part 21 on which a container containing a sample is mounted.
  • a first temperature control unit 3 1 is arranged below the first placement unit 21.
  • a heat radiating section 40 is arranged below the first temperature control section 31.
  • FIG. 3 is a front view of the vicinity of the portion on which the container knife is placed
  • FIG. 4 is a plan view of the vicinity of the portion on which the container knife is placed.
  • a first temperature control unit 31, a second temperature control unit 3 2, and a third temperature control unit 33 are provided on one heat dissipation unit 40. , Are arranged adjacent to each other in this order.
  • the first temperature control unit 3 1, the second temperature control unit 3 2, and the third temperature control unit 33 are cooled on the lower surface side when the upper surface side is heated and on the lower surface when the upper surface side is cooled. It is composed of a Peltier element whose side is heated.
  • the heat dissipation part 40 has a large number of fins formed of a material having a high thermal conductivity such as metal.
  • the heat radiating section 40 promotes heat exchange between the lower surface of the first temperature adjusting section 31, the second temperature adjusting section 32, and the lower surface of the third temperature adjusting section 33 and the ambient air.
  • the first placement unit 21 is arranged on the first temperature control unit 31.
  • the second mounting portion 2 2 is arranged on the second temperature control portion 32.
  • a third mounting portion 23 is arranged on the third temperature control portion 33.
  • the first placing part 21, the second placing part 22 and the third placing part 23 are each formed of a material having a high thermal conductivity such as metal.
  • the upper surface of each of the first placing portion 21, the second placing portion 2 2 and the third placing portion 2 3 has a first depression 2 1 3 and a second depression 2 2 ⁇ 02020/174918 5 ⁇ (: 170?2020/001355
  • a plurality of 2 3 and 3rd recesses 2 3 3 are formed.
  • the first depression 2 1 3, the second depression 2 2 3 and the third depression 2 3 3 are parts where the wells of the container containing the sample are placed.
  • a first thermometer 5 1 is attached to the first mounting portion 21.
  • a second thermometer 5 2 is attached to the second mounting portion 22.
  • a third thermometer 5 3 is attached to the third mounting portion 23.
  • the first thermometer 51, the second thermometer 52, and the third thermometer 53 are each, for example, a thermocouple.
  • the first thermometer 5 1 detects the temperature of the first mounting portion 2 1. Since the first placing part 21 is made of a material having a high thermal conductivity, the temperature of the first placing part 21 will not be affected by the temperature of the container placed on the first placing part 21. It can be regarded as the temperature of the contained sample. The same applies to the temperature of the second mounting portion 22 and the temperature of the third mounting portion 23.
  • the first mounting portion 21 has a plurality of first depressions 21.
  • first depressions 2 1 3 are arranged in a matrix. Specifically, four rows of eight first depressions 2 1 3 are arranged. The intervals between the first depressions 2 1 3 arranged in the row direction are equal at any position. Also, the intervals between the first depressions 2 1 3 arranged in the row direction are equal at any position.
  • the second support 2 2, a plurality of second recesses 2 2 3 are arranged in a matrix. Specifically, four rows of eight second recesses 2 2 3 are arranged. The intervals between the second depressions 2 2 3 arranged in the row direction are equal at any position. Also, the intervals between the second depressions 2 2 3 arranged in the row direction are equal at any position.
  • a plurality of third recesses 2 3 3 are arranged in a matrix on the third mounting portion 23. Specifically, four rows of eight third depressions 2 3 3 are arranged. The intervals between the third depressions 2 3 3 arranged in the row direction are equal at any position. Also, the intervals between the third depressions 2 3 3 arranged in the row direction are equal at any position.
  • first depression 2 1 3 and the second depression 2 2 3 which are adjacent to each other (Fig. 4 ⁇ 02020/174918 6 In the case of (: 170?2020/001355, the distance between the first depression 2 1 3 in the rightmost row and the second depression 2 2 3 in the leftmost row) is adjacent in the row direction. The distance between the first depressions 2 1 3 and the distance between the second depressions 2 2 3 are equal to each other.
  • the second recess 2 2 3 and the third recess 2 3 3 that are adjacent to each other Is equal to the distance between the second depressions 2 2 3 adjacent to each other in the row direction and the distance between the third depressions 2 3 3 adjacent to each other.
  • Figs. 5 and 5 are a plan view and a side view, respectively, showing an example of a container knife used with the nucleic acid amplification device 1 according to the present embodiment.
  • the container knife has eight wells in which the sample is stored, and the eight wells are arranged in a row at equal intervals.
  • the spacing between wells adjacent to each other is the spacing between the first depressions 2 1 3 adjacent to each other in the row direction, the spacing between the second depressions 2 2 3 and the spacing between the third depressions 2 3 3 Is equal to
  • the container knife shown in FIGS. 58 and 5 is placed on the first placing portion 21, the second placing portion 22 and the third placing portion 23. Can be placed one by one. It goes without saying that it is not always necessary to mount four container knives on each mounting part. If necessary, 3 or less container knives may be placed on each mounting part, or no container knives may be mounted on either mounting part.
  • FIG. 6 and FIG. 6 are a plan view and a side view showing another example of the container knife that can be used with the nucleic acid amplification device 1 according to the present embodiment. ⁇ 02020/174918 7 ⁇ (: 170?2020/001355
  • the container has wells in which the samples are placed in a matrix. Specifically, it has 12 rows of 8 wells.
  • the spacing between the wells that are adjacent to each other in the column direction is equal to the spacing between the first depressions 2 1 3 that are adjacent to each other at any position. Also, the distance between the wells that are adjacent to each other in the row direction is equal to the distance between the first depressions 2 1 3 that are adjacent to each other in the row direction at any position.
  • One container knife shown in FIGS. 6 and 6 can be placed on the second placing portion 22 and the third placing portion 23. That is, the nucleic acid amplification device 1 according to the present embodiment can be used with the container knife shown in FIGS. 68 and 6 if necessary.
  • FIG. 7 is a time chart 100 showing the temperature cycle at ⁇ .
  • the horizontal axis represents time and the vertical axis represents sample temperature.
  • the control unit 17 controls the first temperature control unit 31 to raise the temperature of the upper surface of the first temperature control unit 3 1. At the same time, the temperature of the sample in the container rises, and [ ⁇ ] shifts to the enzyme activation step 10 1. In the enzyme activation step 101, the temperature of the sample reaches a predetermined temperature (for example, 95 ° ⁇ ).
  • the state where the temperature of the sample is kept constant continues for a predetermined time. ⁇ 02020/174918 8 ⁇ (: 170?2020/001355
  • control unit 17 controls the first temperature adjusting unit 31 to reduce the temperature on the upper surface side of the first temperature adjusting unit 31. At the same time, the temperature of the sample in the container falls.
  • a predetermined temperature for example, 55°
  • the annealing step 103 is completed when a predetermined time (for example, 10 seconds) elapses while the temperature of the sample is kept constant after shifting to the annealing step 103.
  • control unit 17 controls the first temperature adjustment unit 31 to raise the temperature on the upper surface side of the first temperature adjustment unit 3 1. At the same time, the temperature of the sample in the container rises.
  • the process proceeds to the extension reaction step 104.
  • a predetermined temperature for example, 72°
  • the temperature of the sample is kept constant for a predetermined time (for example, 20 seconds).
  • the optical measurement device 13 performs optical measurement. Therefore, the extension reaction step 104 is also an optical measurement step.
  • a cycle 100000 is constituted by the heat denaturation step 102, the annealing step 103, and the extension reaction step 104. This cycle 100,000 is repeated multiple times, for example 40 times, to complete one sample.
  • the nucleic acid amplification device 1 since it is possible to start the ⁇ of the next sample before the ⁇ [3 ⁇ 4 of one sample is completed, a plurality of samples can be obtained in a short time. of It can be performed.
  • the nucleus according to the present embodiment ⁇ 02020/174918 9 ⁇ (: 170?2020/001355
  • the sample placed on the first placing part 21 is referred to as a first sample.
  • the sample placed on the second placing part 22 is referred to as a second sample.
  • a sample placed on the third placing part 23 is referred to as a third sample.
  • FIG. 8 is a time chart showing a temperature cycle of control executed by the nucleic acid amplification device 1 according to one embodiment.
  • the horizontal axis is the time and the vertical axis is the sample temperature.
  • the time chart 110 is a time chart showing the temperature cycle of the first sample at ⁇ .
  • the time chart 120 is a time chart showing the temperature cycle of the second sample at 0.
  • the time chart 1300 is a time chart showing the temperature cycle at 0 of the third sample.
  • FIGS. 9 to 90 are display examples on the operation display unit 16 provided in the nucleic acid amplification device 1 according to the embodiment, respectively.
  • the cover 12 is opened, and the first sample at room temperature is placed on the first placing part 21. Then, the cover 12 is closed and locked, and the operation display unit 16 is operated to set the condition of ⁇ for the first sample.
  • the temperature of the first sample is room temperature, which means that
  • FIG. 9-8 A display example of the operation display unit 16 at this time is shown in FIG. In Figure 9-8, a polygonal line imitating Time Chart 110 is shown.
  • condition setting of ⁇ of the first sample can be executed, for example, by pressing the "edit" button shown in Fig. 9. It should be noted that the above-mentioned condition setting of the first sample may be performed before the first sample is mounted on the first mounting part 21.
  • the control of the temperature control unit 31 of 1 is started, that is, the temperature of the first sample is started. Then, the controller 17 performs the first temperature adjustment according to a predetermined sequence. ⁇ 02020/174918 10 box (: 170?2020/001355
  • Heat or cool part 3 Heat or cool part 3 1.
  • the enzyme activation step 1 1 1 is performed, followed by the heat denaturation step 1 1 2, annealing step 1 1 3 and extension reaction step 1 1 1.
  • Cycle 1 1 0 ⁇ 3 including 4 is repeated.
  • FIG. 9 shows a display example of the operation display unit 16 when the first sample is subjected to the heat denaturation step 11 2.
  • FIG. 90 shows a display example of the operation display unit 16 of the second sample when the condition [0] is set.
  • the polygonal line imitating Time Chart 1100 is shown on the upper side
  • the polygonal line imitating Time Chart 120 is shown on the lower side.
  • the "pause" button on the operation display unit 16 cannot be pressed.
  • the temperature of the second sample at this time is room temperature.
  • the cover 12 When the second sample is placed on the second placing section 22, the cover 12 is opened, so that the temperature inside the housing 11 is lowered.
  • annealing step 113 is the coldest process in the first sample, ⁇ . Therefore, the adverse effect of opening the cover 12 and lowering the temperature inside the housing 11 is minimized.
  • Fig. 8 shows that the 0 cycle 1 1 1 00 of the first sample and the 0 cycle 1 2 0 0 of the second sample are synchronized.
  • control unit 17 controls the temperature of the first sample to increase and the temperature of the first sample to decrease while the temperature of the first sample increases.
  • the first temperature control unit 3 1 and the second temperature control unit 3 2 are controlled so that the temperature of the second sample decreases when the temperature is high.
  • the control unit 17 determines that the temperature of the first temperature adjustment unit 3 1 is the temperature at which the first sample is subjected to the heat denaturation step 1 1 2.
  • the temperature of the second temperature control unit 3 2 should be the temperature at which the second sample is subjected to the heat denaturation step 1 2 2 so that the temperature of the first temperature control unit 3 1 and the second temperature control unit 3 2 Control 3 2
  • the control unit 17 determines that the temperature of the first temperature adjusting unit 3 1 is the temperature at which the first sample is subjected to the annealing step 1 1 3.
  • the temperature of the second temperature control unit 32 is set to the temperature when the second sample is subjected to the annealing process 1 23, so that the temperature of the first temperature control unit 31 ⁇ 02020/174918 12 ((170?2020/001355
  • control unit 17 controls the temperature of the first temperature adjusting unit 31 to be the temperature at which the first sample is subjected to the extension reaction step 1 14.
  • the temperature of the second temperature control unit 3 2 is adjusted to the temperature at which the second sample is subjected to the extension reaction step 1 2 4, so that the first temperature control unit 3 1 and the second temperature control unit 3 2 Control part 32.
  • the display of the operation display unit 16 becomes, for example, the display shown in FIG. FIG. 9 shows that 0 of the first sample is in the heat denaturation step 1 12 and 0 of the second sample is in the heat denaturation step 12 2.
  • the optical measurement device 13 irradiates the first sample and the second sample with excitation light at one time, and Detect fluorescence at once.
  • the optical measuring device 13 has a plurality of photodetecting elements arranged two-dimensionally. Therefore, the fluorescence emitted from the first sample and the second sample arranged in a matrix can be detected simultaneously for each sample separately.
  • the fluorescence detection result is analyzed by the controller 17.
  • the information on the amplification status of the nucleic acid contained in the first sample and the information on the amplification status of the nucleic acid contained in the second sample, which are obtained as a result of the analysis, are as follows: ⁇ for the first sample and ⁇ for the second sample. During operation, it is displayed on the operation display box ⁇ 16 at all times or at the request of the user. Alternatively, this information can be obtained from the first sample ⁇ 02020/174918 13 ⁇ (: 170?2020/001355
  • the display of the operation display unit 16 becomes the state shown in FIG.
  • the nucleic acid amplification device 1 continues the second ⁇ [3 ⁇ 4 until the second ⁇ [3 ⁇ 4 is completed and follows the predetermined sequence to display information on the amplification status of the nucleic acid contained in the second sample. Keep getting.
  • nucleic acid amplification apparatus 1 has been described when the first sample O and the second sample O are performed in parallel.
  • the nucleic acid amplification apparatus 1 according to this embodiment can further perform the third sample ⁇ in parallel.
  • the operation of the nucleic acid amplification device 1 when performing is described.
  • the control unit 17 controls the first temperature control unit 3 1 and the second temperature control unit 3 2. If a predetermined operation is performed on the operation display unit 16 while the operation is in progress (for example, the “Add reaction” button in Figure 9 is pressed) It is possible to set the conditions of.
  • the operation display unit 16 displays the broken line of the time chart 1 10 and the broken line of the time chart 1 20. A line that shows Time Chart 1300 is displayed below.
  • control unit 17 controls the first temperature control unit 31, the second temperature control unit 32, and the third temperature control unit 33 to operate the 1st cycle 11 of the first sample. Control so that the ⁇ cycle, the ⁇ cycle of the second sample 1 200, and the ⁇ cycle of the 3rd sample 1300 are synchronized.
  • Fig. 8 shows that 1 200, and the cycle of 130 of the third sample 1300 are synchronized. As shown in FIG. 8, at the timing 204, the extension reaction step 1 1 4 of the first sample, the extension reaction step 1 2 4 of the second sample, and the extension reaction step 1 3 of the third sample 1 3 4 will be carried out simultaneously.
  • the optical measurement device 13 irradiates the first sample, the second sample, and the third sample with excitation light at a time, and at the same time, the first sample, Fluorescence is detected at once from the second and third samples.
  • the optical measuring device 13 has a plurality of photodetecting elements arranged two-dimensionally. Therefore, the fluorescence emitted from the first sample, the second sample, and the third sample arranged in a matrix can be detected simultaneously for each sample separately.
  • the ⁇ of the first sample is in the extension reaction step 1 1 4
  • the ⁇ of the second sample is in the extension reaction step 1 2 4
  • the ⁇ of the third sample is in the extension reaction step 1 3 4.
  • the optical measurement device 13 executes the fluorescence detection from each sample each time the timing 204 is reached.
  • the fluorescence detection result is analyzed by the controller 17.
  • the analysis provides information on the amplification status of the nucleic acid contained in the first sample, information on the amplification status of the nucleic acid contained in the second sample, and information on the amplification status of the nucleic acid contained in the third sample. ..
  • These pieces of information can be used at any time during the operation of ⁇ [3 ⁇ 4 for the first sample, ⁇ [3 ⁇ 4 for the second sample and ⁇ for the third sample, or at any time during the operation of the user. Displayed on display unit 16.
  • the information may be collected from the user each time the information is obtained while ⁇ [3 ⁇ 4 of the first sample, ⁇ of the second sample and ⁇ of the third sample are being performed.
  • On request ⁇ 02020/174918 15 ⁇ (: 170?2020/001355
  • the nucleic acid amplification device 1 Even when the first 0 is completed, the nucleic acid amplification device 1 continues the second ⁇ [3 ⁇ 4 to the second sample according to a predetermined sequence until the second ⁇ is completed. Continue to obtain information on the amplification status of the included nucleic acids. Also, the first Even if is completed, the nucleic acid amplification device 1 continues the third ⁇ [3 ⁇ 4 until the third ⁇ [3 ⁇ 4 is completed, according to a predetermined sequence, and Continue to get information.
  • the nucleic acid amplification device 1 Even if the second ⁇ is completed in addition to the first ⁇ , the nucleic acid amplification device 1 continues the third ⁇ [3 ⁇ 4 according to a predetermined sequence until the third ⁇ is completed. And continue to obtain information on the amplification status of the nucleic acid contained in the third sample.
  • nucleic acid amplification device 1 of the present embodiment a plurality of samples Can be started sequentially. Therefore, it is possible to efficiently perform ⁇ of a plurality of samples in a short time.
  • FIG. 10 is a front view of the vicinity of the portion where the container is placed in the nucleic acid amplification device 1 according to another embodiment, and is a view corresponding to FIG.
  • the first heat radiation section 41 is arranged below the first temperature control section 31.
  • the second heat radiation section 42 is arranged below the second temperature control section 32.
  • the third heat dissipation section 43 is arranged below the third temperature control section 33.
  • the first placing part 21, the first temperature adjusting part 31 and the first heat releasing part 41, the second placing part 22 and the second temperature adjusting part 41 are arranged.
  • a heat insulating material between the second mounting section 23, the third mounting section 23, the third temperature control section 33, and the third heat dissipating section 43.
  • the nucleic acid amplification device according to the present invention is not limited to the above embodiments.
  • the optical measurement device 13 detects fluorescence individually and continuously from each of a plurality of samples arranged in a matrix by scanning one or a plurality of photodetection elements. You may. Also in this case, the optical measuring device 13 can detect light emitted from a plurality of samples, for example, the first sample and the second sample, at one time.
  • the second sample when the temperature of the first temperature control unit 31 is the temperature at which the first sample is subjected to the annealing step 113, the second sample is set to the casing.
  • the cover 1 2 was unlocked to fit inside 11.
  • the unlocking of the cover 12 may be performed at another timing. For example, even if the cover 1 2 is unlocked when the temperature 3 1 of the first temperature control unit is decreasing toward the temperature at which the first sample is subjected to the annealing step 1 13 3. Good.
  • the second sample can be set at an early timing, and the temperature of the first temperature control unit 31 and thus the temperature of the first sample can be lowered more quickly. You can
  • the unlocking of the cover 12 may be performed at another timing.
  • the control unit 17 may unlock the cover 1 2. Yes.
  • the additional sample can be set at a relatively arbitrary timing, which improves user pity. Unlocking with such a timing will reduce the temperature drop of the sample placed in the housing. ⁇ 02020/174918 17 ⁇ (: 170?2020/001355
  • the nucleic acid amplification device 1 may be provided with a notifying unit for notifying by sound or light that 0 of one of the samples is completed. If the nucleic acid amplification device 1 is equipped with a notification unit, The user can immediately know the inspection result of the completed sample.
  • the mounting unit includes a first mounting unit 21 and a second mounting unit 2
  • the second and third mounting portions 23 are divided into three in the column direction.
  • the placement section may be divided into a plurality of rows.
  • the container may be composed of one well or may be composed of a plurality of wells connected at equal intervals in the row direction.
  • the first placing part 21, the second placing part 22 and the third placing part 23 may be further divided into a plurality of columns or rows. As a result, the temperature distribution of each mounting portion can be controlled more finely.
  • the nucleic acid amplification device 1 according to the present invention is not limited to the so-called 3 step ⁇ consisting of three steps of a thermal denaturation step, an annealing step and an extension reaction step, and an annealing and extension reaction are performed in one step. It goes without saying that the so-called 2 steps can be performed.
  • the present invention contributes to improving the efficiency of inspection in a nucleic acid amplification device such as a real-time device, and its industrial applicability is great. Explanation of symbols ⁇ 02020/174918 18 ?01/1?2020/001355

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Abstract

Dispositif d'amplification d'acide nucléique comprenant: une première unité de régulation de température pour chauffer et refroidir un premier échantillon contenant un acide nucléique; une seconde unité de régulation de température pour chauffer et refroidir un second échantillon contenant un acide nucléique; et une unité de commande pour commander la première unité de régulation de température et la seconde unité de régulation de température et démarrer la commande de la seconde unité de régulation de température au cours de la commande de la première unité de régulation de température.
PCT/JP2020/001355 2019-02-27 2020-01-16 Dispositif d'amplification d'acide nucléique Ceased WO2020174918A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011501665A (ja) * 2007-10-12 2011-01-13 レオニックス，インコーポレイテッド 統合型マイクロ流体デバイスおよび方法
WO2011138925A1 (fr) * 2010-05-07 2011-11-10 株式会社日立ハイテクノロジーズ Amplificateur d'acides nucléiques et dispositif d'inspection d'acides nucléiques l'employant
JP2011239688A (ja) * 2010-05-14 2011-12-01 Hitachi High-Technologies Corp 核酸増幅装置及び核酸増幅方法
WO2012011379A1 (fr) * 2010-07-21 2012-01-26 株式会社日立ハイテクノロジーズ Procédé de détection d'une amplification d'acide nucléique dans un échantillon et dispositif pour ce faire
JP2017121245A (ja) * 2011-04-15 2017-07-13 ベクトン・ディキンソン・アンド・カンパニーＢｅｃｔｏｎ， Ｄｉｃｋｉｎｓｏｎ Ａｎｄ Ｃｏｍｐａｎｙ 走査リアルタイムマイクロ流体熱サイクラーと同期熱サイクリング及び走査光学検出の方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011501665A (ja) * 2007-10-12 2011-01-13 レオニックス，インコーポレイテッド 統合型マイクロ流体デバイスおよび方法
WO2011138925A1 (fr) * 2010-05-07 2011-11-10 株式会社日立ハイテクノロジーズ Amplificateur d'acides nucléiques et dispositif d'inspection d'acides nucléiques l'employant
JP2011239688A (ja) * 2010-05-14 2011-12-01 Hitachi High-Technologies Corp 核酸増幅装置及び核酸増幅方法
WO2012011379A1 (fr) * 2010-07-21 2012-01-26 株式会社日立ハイテクノロジーズ Procédé de détection d'une amplification d'acide nucléique dans un échantillon et dispositif pour ce faire
JP2017121245A (ja) * 2011-04-15 2017-07-13 ベクトン・ディキンソン・アンド・カンパニーＢｅｃｔｏｎ， Ｄｉｃｋｉｎｓｏｎ Ａｎｄ Ｃｏｍｐａｎｙ 走査リアルタイムマイクロ流体熱サイクラーと同期熱サイクリング及び走査光学検出の方法

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