WO2023195945A1 - Dispositif de pcr isotherme - Google Patents

Dispositif de pcr isotherme Download PDF

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Publication number
WO2023195945A1
WO2023195945A1 PCT/TR2022/050940 TR2022050940W WO2023195945A1 WO 2023195945 A1 WO2023195945 A1 WO 2023195945A1 TR 2022050940 W TR2022050940 W TR 2022050940W WO 2023195945 A1 WO2023195945 A1 WO 2023195945A1
Authority
WO
WIPO (PCT)
Prior art keywords
circuit board
pcr device
isothermal pcr
heating
sample
Prior art date
Application number
PCT/TR2022/050940
Other languages
English (en)
Inventor
Huseyin AKCAY
Fatih Mehmet DUMANOGULLARI
Original Assignee
Yonca Teknoloji Muhendislik Ve Elektronik Hiz. Ltd. Sti.
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 Yonca Teknoloji Muhendislik Ve Elektronik Hiz. Ltd. Sti. filed Critical Yonca Teknoloji Muhendislik Ve Elektronik Hiz. Ltd. Sti.
Publication of WO2023195945A1 publication Critical patent/WO2023195945A1/fr

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/12—Specific details about manufacturing devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0654—Lenses; Optical fibres
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1827—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/06—Test-tube stands; Test-tube holders
    • 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 invention relates to an isothermal PCR device.
  • the invention relates in detail to an isothermal PCR device having a compact structure.
  • PCR polymerase chain reaction
  • the PCR test devices include light sources to emit light on the sample and various focusing elements to detect the light emitted from the sample, filters and photo sensors.
  • PCR test devices designed for the simultaneous performance of multiple tests in the present art comprise a motion mechanism for moving the detector around the test tubes for the measurement of light emitted from the sample.
  • it includes structures such as a gear mechanism with filters for positioning the filters that filter at different wavelengths between the detector and the reflected light. The design, operation, use and maintenance costs of mobile mechanisms for devices of these sizes are considerably increased.
  • the present invention relates to an isothermal PCR device to eliminate the above-mentioned disadvantages and bring new advantages to the relevant technical field.
  • An object of the invention is to provide a compact isothermal PCR device in which the volume it occupies is reduced, that is, its size is reduced.
  • An object of the invention is to provide a PCR device in which the rate of reaching the appropriate temperature and the test speed is increased.
  • An object of the invention is to provide an isothermal PCR device with increased ease of assembly.
  • Another object of the invention is to provide an isothermal PCR device with reduced production cost.
  • the present invention relates to an isothermal PCR device for testing a test tube containing a sample and a reagent mixture to realize all the purposes that will emerge from the abovementioned and the following detailed description. Accordingly, it is characterized in that it comprises a first circuit board, a second circuit board positioned on the first circuit board at a predetermined distance between the said first circuit board; the said second circuit board comprises at least one opening for placing the test tube; it comprises at least one light source positioned on the second circuit board to send light to the sample in each test tube disposed in the said opening; it comprises at least one detector positioned to face each tube on the first circuit board to detect the light emitted or scattered from the sample; and a control unit configured to control the operation of the said detector with the said light source and detect the presence of at least one analyte in the sample according to the signal received from the detector.
  • the ease of production and assembling of the isothermal PCR device is increased.
  • a possible embodiment of the invention is characterized in that it comprises a heating block with openings provided mutually with the openings on the second circuit board for the placement of the said test tube and for keeping the environment in which the test tube is located at a certain temperature; at least one heating element for heating the said heating block; and a temperature sensor for measuring the ambient temperature in which the test tube is located.
  • a heating block with openings provided mutually with the openings on the second circuit board for the placement of the said test tube and for keeping the environment in which the test tube is located at a certain temperature
  • at least one heating element for heating the said heating block
  • a temperature sensor for measuring the ambient temperature in which the test tube is located.
  • Another possible embodiment of the invention is characterized in that the first circuit board is mountable by placing a heating block on the second circuit board and the second circuit board. In this way, the ease of installation of the isothermal PCR device is increased. Thus, it is ensured that the isothermal PCR device has a compact structure.
  • Another possible embodiment of the invention is characterized in that the said heating block is made of aluminum material.
  • the weight and cost of the isothermal PCR device are reduced.
  • Another possible embodiment of the invention is characterized in that the said heating element and the said temperature sensor are provided on the second circuit board.
  • the heating element relates to the heating block.
  • the temperature sensor is enabled to measure the temperature of the area remaining in the heating block.
  • Another possible embodiment of the invention is characterized in that the said heating elements are provided in multiple numbers at certain intervals on the edges of the second circuit board. Thus, it is ensured that the heating block is heated homogeneously.
  • the heating block comprises a heating duct associated with the heating elements.
  • the heat loss of the heating block is reduced.
  • the heating block has as many openings as the number of openings the second circuit board has; the said openings are mutually provided.
  • the test tube is placed in the isothermal PCR device.
  • Another possible embodiment of the invention is characterized in that the detector positioned on the first circuit board is provided with the number of openings provided on the second circuit board and the heating block; the detectors are provided reciprocally with the said openings.
  • Another possible embodiment of the invention is characterized in that the openings provided on the second circuit board are configured to allow at least partial passage of the test tube. Thus, the light emitted from the light source located on the second circuit board is prevented from reaching the detector directly.
  • control unit is configured to control the operation of the heating elements according to the temperature signal received from the temperature sensor.
  • Another possible embodiment of the invention is characterized in that the said heating element is the resistor.
  • Another possible embodiment of the invention is characterized in that it comprises input output pins for the electrical connection of the first circuit board and the second circuit board.
  • Another possible embodiment of the invention is characterized in that the said input output pins are configured to sit on the first circuit board of the second circuit board.
  • Another possible embodiment of the invention is characterized in that the detector provided on the first circuit board is positioned to make a right angle with the light sent on the sample by means of the light source.
  • the detector does not receive direct light from the light source and the light reflected from the sample tube is minimized. In this way, light noise is reduced in the measurement of radiations such as fluorescence and phosphorescence made by the sample and/or scattering from the sample.
  • Another possible embodiment of the invention is characterized in that it comprises a filter element for filtering light of a certain wavelength between the first circuit board and the second circuit board.
  • a filter element for filtering light of a certain wavelength between the first circuit board and the second circuit board.
  • Another possible embodiment of the invention is characterized in that it comprises a supporting block made of thermal insulating material and configured to surround the first circuit board, the second circuit board and the heating block.
  • a supporting block made of thermal insulating material and configured to surround the first circuit board, the second circuit board and the heating block.
  • Another possible embodiment of the invention is characterized in that it comprises a cover made of thermal insulating material and configured to be closed on the said supporting block; the test tubes placed in the first circuit board, the second circuit board, the heating block and the heating block are configured to remain within the said supporting block with the said cover.
  • thermal insulation of the isothermal PCR device from the external environment is provided.
  • Another possible embodiment of the invention is characterized in that it comprises a sealing element so as to enclose at least one of the said cover or the said supporting block. Thus, heat dissipation between the cover and the supporting block is prevented.
  • Another possible embodiment of the invention is characterized in that the said sealing element is provided on the supporting block. Thus, it is ensured that the cover is closed compressively on the supporting block.
  • Another possible embodiment of the invention is characterized in that it comprises a user interface; the said control unit is configured to indicate the test result of the said control unit at the said user interface. Thus, it is ensured that the results of the tests can be examined by the users.
  • Figure 1 shows a perspective view of the isothermal PCR device.
  • Figure 2 shows a perspective view of the internal structure of the isothermal PCR device.
  • Figure 3 shows a cross-sectional view of the exploded structure of the isothermal PCR device.
  • Figure 4 shows a view of the uncapped structure of the isothermal PCR device.
  • Figure 5 shows a view of the covered structure of the isothermal PCR device.
  • Figure 6 shows a representative view of the heating block.
  • the invention relates to an isothermal PCR device (10) for detecting the presence of at least one analyte in a sample.
  • the isothermal PCR device (10) mentioned herein detects the presence of the analyte by the isothermal PCR test method known in the art.
  • the said sample may be placed in the isothermal PCR device (10) together with a reagent mixture by means of a test tube (300).
  • the said analyte may be a content in clinical chemistry or a chemical species or a chemical content targeted in an analytical procedure.
  • the said analyte may be a microscopic infectious virus capable of infecting living cells and thus replicating them.
  • the isothermal PCR device (10) comprises a heating block (100) for maintaining the environment in which the test tube (300) is located at a certain temperature.
  • the said heating block (100) has at least one opening (140) for inserting the test tube (300).
  • the heating block (100) has a plurality of openings (140). It ensures that the test tubes placed in the heating block (100) can be tested at a certain temperature.
  • the isothermal PCR device (10) comprises at least one light source (121) positioned to send light on the sample in the test tube (300). The said light source (121 ) sends light on the sample, allowing light to be scattered from the sample and the sample to irradiate.
  • the isothermal PCR device (10) comprises at least one detector (111) for detecting the radiation of the sample and/or the light scattered from the sample through the light source (121).
  • the said detector (111 ) senses the wavelength and/or intensity of the light.
  • a photosensor known in the art may be used as the detector (111).
  • the detector (111) is positioned to make a certain angle with the light sent from the light source (121) onto the sample.
  • the detector (111 ) is positioned to make a right angle with the light sent on the sample by the light source (121 ).
  • the light source (121) is configured to send light to a side face of the test tube (300); the detector (111 ) is configured to be located under the test tube (300).
  • the isothermal PCR device (10) includes a control unit (200).
  • the said control unit (200) enables simultaneous operation of the light source (121) and the detector (111).
  • the control unit (200) is configured to detect the presence of at least one analyte in the sample according to the signal it receives from the detector (111).
  • the isothermal PCR device (10) includes a first circuit board (110) for positioning the detector (111 ).
  • the isothermal PCR device (10) includes a second circuit board (120) for positioning the light source (121).
  • the said second circuit board (120) is disposed on the first circuit board (110) so as to maintain a certain distance between the said first circuit board (110).
  • the first circuit board (110) and the second circuit board (120) comprise input output pins (130) to enable them to be electrically connected.
  • the input output pins (130) are configured to fit on the second circuit board (120) and the first circuit board (110).
  • the isothermal PCR device (10) is configured so that the second circuit board (120) remains within the heating block (100) and the first circuit board (110) is located at the bottom of the heating block (100). In this way, it is ensured that the isothermal PCR device (10) has a compact structure. Thus, the ease of production and assembly of the isothermal PCR device (10) is increased.
  • the first circuit board (110) includes connection openings (140) at the corner portions of the second circuit board (120) and the heating block (100).
  • the said connection openings (140) provide a fixed attachment of the first circuit board (110), the second circuit board (120), and the heating block (100).
  • the isothermal PCR device (10) comprises at least one heating element (122) for heating the said heating block (100).
  • the said heating element (122) is at least partially associated with the heating block (100).
  • the heating element (122) may be positioned on the first circuit board (110) or the second circuit board (120). In the preferred embodiment of the invention, the heating element (122) is positioned on the second circuit board (120). In this way, the heating element (122) remains in the heating block (100).
  • the heating block (100) is selected from the thermally conductive material to provide homogeneous heat dissipation.
  • the said thermally conductive material can be made of materials known in the art such as iron, copper, and aluminum.
  • the isothermal PCR device (10) comprises a heating block (100) made of aluminum material.
  • the heating block (100) includes a heating duct (101) to provide homogeneous heat dissipation.
  • the heating duct (101) is provided to reduce the air gap in the heating block (100).
  • the heating duct (101 ) is provided to be at least partially associated with the heating element (122).
  • the isothermal PCR device (10) comprises a plurality of provided heating elements (122).
  • the heating elements (122) are positioned on the second circuit board (120).
  • the heating elements (122) are positioned on the second circuit board (120) in such a way that they are arranged at regular intervals. In this way, it is ensured that the heating block (100) is heated homogeneously.
  • the said heating elements (122) are electrical components that convert electrical energy into heat energy.
  • the resistance is used as the heating element (122).
  • the isothermal PCR device (10) comprises a temperature sensor (123) for measuring the ambient temperature at which the test tube (300) is located.
  • the said temperature sensor (123) is positioned in the heating block (100).
  • the temperature sensor (123) is preferably positioned on the second circuit. In this way, temperature measurement in the heating block (100) is provided.
  • the control unit (200) controls the operation of the heating elements (122) according to the temperature information it receives from the temperature sensor (123).
  • the second circuit board (120) comprises an opening (140) equal to the number of openings (140) that the heating block (100) has.
  • the first circuit board (110) comprises the heating block (100) and the detector (111 ) as many as the number of openings (140) that the second circuit board (120) has.
  • the said openings (140) and the detectors (111) are provided mutually.
  • the openings (140) of the second circuit board (120) are configured to allow at least partial passage of the test tube (300). In this way, the test tubes placed in the openings (140) of the heating block (100) pass partially through the openings (140) of the second circuit board (120) remaining in the heating block (100).
  • the light sources (121) on the second circuit board (120) send light on the sample from the side face of the test tube (300).
  • the detectors (111 ) located on the first circuit board (110) under the openings (140) on the second circuit board (120) detect the light scattered over the sample.
  • the isothermal PCR device (10) includes a filter element (150) such that it lies between the first circuit board (110) and the second circuit board (120).
  • the said filter element (150) provides filtering of light at a certain wavelength.
  • the filter element (150) provides filtering to pass the light above a certain threshold wavelength.
  • the threshold wavelength of the filter element (150) is selected to be greater than the wavelength of the light emitted by the light source (121).
  • the light source (121) emits light at a wavelength of 470 nm.
  • the filter element (150) used herein filters light up to a wavelength of 500 nm and allows light higher than a wavelength of 500 nm to pass through.
  • the detector (111) is prevented from detecting the light emitted from the light source (121).
  • the detector (111) detects only the radiation (emission) of the sample.
  • the filter element (150) can be selected as a light filter with any filtering feature known in the art.
  • the light source (121) may be selected as a light source (121) to emit light at any wavelength.
  • a filter element (150) specific to the type of an analyte known to emit at what wavelength may be used. For example, a certain analyte presence in the sample can be detected using a filter element (150) that passes the 500-530 nm band.
  • the isothermal PCR device comprises a supporting block (160) made of thermally insulating material.
  • the said supporting block (160) is configured to surround the first circuit board (110), the second circuit board (120), and the heating block (100). In this way, the heat provided in the heating block (100) is insulated.
  • the isothermal PCR device (10) also includes a cover (170) made of thermally insulating material.
  • the said cover (170) is configured to close on the supporting block (160). By closing the cover on the supporting block (160), the first circuit board (110), the second circuit board (120), the heating block (100), and the test tubes placed in the heating block (100) remain in the supporting block (160) with the cover (170).
  • the isothermal PCR device (10) includes a sealing element (161 ) to enclose at least one of the cover (170) or the supporting block (160).
  • the said sealing element (161 ) prevents heat loss between the cover (170) and the supporting block (160).
  • the sealing element (161 ) is preferably provided on the supporting block (160).
  • the isothermal PCR device (10) comprises a user interface (180).
  • the control unit (200) is configured to display the test results to the user via the said user interface (180).

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

L'invention concerne un dispositif de PCR isotherme (10) destiné à tester un tube à essai (300) comprenant un échantillon et un mélange de réactifs. En conséquence, ledit dispositif de la présente invention est caractérisé en ce qu'il comprend une première carte de circuit imprimé (110) ; une seconde carte de circuit imprimé (120) positionnée sur la première carte de circuit imprimé (110) à une distance prédéterminée de ladite première carte de circuit imprimé (110), ladite seconde carte de circuit imprimé (120) présentant au moins une ouverture pour placer le tube à essai (300) ; au moins une source de lumière positionnée sur la seconde carte de circuit imprimé (120) pour envoyer de la lumière à l'échantillon dans chaque tube à essai (300) disposé dans ladite ouverture ; au moins un détecteur (111) positionné pour faire face à chaque tube sur la première carte de circuit imprimé (110) pour détecter la lumière émise ou diffusée à partir de l'échantillon ; et une unité de commande (200) conçue pour commander le fonctionnement dudit détecteur (111) avec ladite source de lumière et détecter la présence d'au moins un analyte dans l'échantillon en fonction du signal reçu en provenance du détecteur (111).
PCT/TR2022/050940 2022-04-05 2022-09-02 Dispositif de pcr isotherme WO2023195945A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
TR2022/005362 2022-04-05
TR2022/005362A TR2022005362A2 (tr) 2022-04-05 2022-04-05 İzotermal pcr ci̇hazi

Publications (1)

Publication Number Publication Date
WO2023195945A1 true WO2023195945A1 (fr) 2023-10-12

Family

ID=85160810

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/TR2022/050940 WO2023195945A1 (fr) 2022-04-05 2022-09-02 Dispositif de pcr isotherme

Country Status (2)

Country Link
TR (1) TR2022005362A2 (fr)
WO (1) WO2023195945A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160320381A1 (en) * 2011-09-25 2016-11-03 Theranos, Inc. Systems and methods for multi-analysis
WO2017160839A1 (fr) * 2016-03-14 2017-09-21 Diassess Inc. Dispositifs et procédés de modification de propriétés optiques
WO2018191252A1 (fr) * 2017-04-10 2018-10-18 Cornell University Système et procédé d'amplification isotherme d'acides nucléiques

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160320381A1 (en) * 2011-09-25 2016-11-03 Theranos, Inc. Systems and methods for multi-analysis
WO2017160839A1 (fr) * 2016-03-14 2017-09-21 Diassess Inc. Dispositifs et procédés de modification de propriétés optiques
WO2018191252A1 (fr) * 2017-04-10 2018-10-18 Cornell University Système et procédé d'amplification isotherme d'acides nucléiques

Also Published As

Publication number Publication date
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