WO2023178710A1 - Instrument portable de pcr quantitative fluorescente ultrarapide en temps réel - Google Patents

Instrument portable de pcr quantitative fluorescente ultrarapide en temps réel Download PDF

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Publication number
WO2023178710A1
WO2023178710A1 PCT/CN2022/083373 CN2022083373W WO2023178710A1 WO 2023178710 A1 WO2023178710 A1 WO 2023178710A1 CN 2022083373 W CN2022083373 W CN 2022083373W WO 2023178710 A1 WO2023178710 A1 WO 2023178710A1
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WO
WIPO (PCT)
Prior art keywords
chip
temperature control
reaction
heating
sampling hole
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PCT/CN2022/083373
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English (en)
Chinese (zh)
Inventor
沈照淋
周志图
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无锡百泰克生物技术有限公司
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Publication of WO2023178710A1 publication Critical patent/WO2023178710A1/fr

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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
    • C12M1/02—Apparatus for enzymology or microbiology with agitation means; with heat exchange means
    • 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 utility model relates to the field of biological detection technology, in particular to a portable ultra-fast real-time fluorescence quantitative PCR instrument.
  • PCR instrument also known as PCR gene amplifier, PCR nucleic acid amplifier, polymerase chain reaction nucleic acid amplifier, is an instrument that uses PCR (Polymerase chain reaction, polymerase chain reaction) technology to amplify specific DNA.
  • PCR Polymerase chain reaction, polymerase chain reaction
  • PCR testing has become the "gold standard" for clinical diagnosis.
  • the utility model In order to overcome the shortcomings of the background technology, the utility model discloses a portable ultra-fast real-time fluorescence quantitative PCR instrument.
  • the portable ultra-fast real-time fluorescence quantitative PCR instrument disclosed by the utility model includes:
  • the PCR reaction chip includes a chip base, a reaction tank is provided on the chip base, and the surface of the reaction tank is covered with a bottom film to form a reaction channel with the reaction tank;
  • the chip mounting module includes a chip mounting base plate, the mounting base plate is provided with a guide slot, and the PCR reaction chip is placed in the guide slot;
  • the temperature-controlled floating module includes a temperature control device.
  • the end of the temperature control device is provided with a heating and cooling piece and a temperature sensor.
  • the heating and cooling piece is provided with a heating and cooling carrier plate corresponding to the PCR reaction chip.
  • the heating and cooling carrier plate There are thermal insulation pressure plates on both sides, which are pressed on the temperature control device through the thermal insulation pressure plates;
  • the optical module includes a switching device and a position sensor; a through hole is provided on the back of the chip mounting base plate as part of the optical path for excitation light and fluorescence to pass through.
  • the chip base is provided with a cylindrical sampling hole, the bottom opening of the cylindrical sampling hole is connected with one end of the reaction channel to form a sampling channel, and the upper end of the cylindrical sampling hole wall has a hole through which The hole wall extends and communicates with the other end of the reaction channel to form a gas outlet channel.
  • the cylindrical sampling hole is threaded and closed by a sampling hole cover.
  • the sampling hole cover is provided with an extrusion head corresponding to the position of the cylindrical sampling hole. As the sampling hole cover rotates, the extrusion head squeezes the sample into the reaction channel. The operation is simple and does not require complex liquid drive components.
  • the extrusion head squeezes the sample, it will squeeze the gas in the sampling channel and reaction tank out of the gas outlet channel.
  • the cylindrical sampling hole is completely closed, the sample is evenly distributed in the reaction chamber.
  • the gas in the sampling channel and the gas outlet The outer end holes of the channel are closed by the sampling hole cover to prevent the reagent from overflowing and causing aerosol contamination during repeated temperature rise and fall, and the sealing performance is reliable and stable.
  • positioning grooves are provided on both sides of the PCR reaction chip, positioning bumps are provided in the guide slot corresponding to the positioning groove, and a first in-position sensor is provided.
  • the chip mounting module also includes a chip sliding bottom plate.
  • the chip sliding bottom plate is provided with a chip guide rail and a second in-position sensor.
  • the driving end is connected to the chip mounting bottom plate to control the movement of the chip along the guide rail and through the second in-position sensor.
  • the sensor identifies the location.
  • the temperature control device is placed on the main temperature control bracket through an L-shaped fixing frame.
  • the temperature control device is connected to the L-shaped fixing frame through the first temperature control guide rail, and the ends are connected through pre-tightened springs to achieve extrusion.
  • Pressure buffering the L-shaped fixed frame is placed on the main temperature control bracket through the second temperature control guide rail, the temperature control device is driven by the second motor to move toward the PCR reaction chip, and a second in-position sensor is provided.
  • the optical module's optical path generates excitation light from an excitation light source, filters out stray light through a filter and collimates the lens, and is reflected to the chip reaction tank through a secondary beam splitter.
  • the generated fluorescence is fluorinated by the secondary beam splitter.
  • the detection device receives, wherein the fluorescence detection device is a photodiode, camera or PMT.
  • the fluorescence detection device can select photodiodes, cameras, and PMTs according to different needs.
  • Figure 1 is a schematic diagram of the overall structure of a case of this utility model
  • Figure 2 is a schematic diagram of the overall structure of Case 2 of the present utility model
  • Figure 3 is a diagram of the initial state of chip insertion in the case of this utility model
  • Figure 4 is a diagram of the initial state of inserting the second chip of the utility model case
  • Figure 5 is an operating status diagram of the utility model case 1;
  • Figure 6 is an operating state diagram of the second case of the utility model
  • Figure 7 is a structural diagram of the PCR reaction chip of the present utility model
  • Figure 8 is a structural diagram of the guide slot of the present utility model
  • FIG. 9 is a structural diagram of the chip mounting module of the present invention.
  • Figure 10 is a structural diagram of the temperature control device of the present utility model
  • Figure 11 is a structural diagram of the temperature-controlled floating module of the present utility model
  • Figure 12 is a structural diagram of an optical module of the utility model case
  • Figure 13 is a structural diagram of the second optical module of the utility model
  • Figure 14 is a schematic diagram of the optical path structure of the utility model case 1;
  • Figure 15 is a schematic diagram of the structure of the second optical path in the case of this utility model.
  • the portable ultra-fast real-time fluorescence quantitative PCR instrument shown in Figure 1-6 includes: PCR reaction chip 1, chip installation module 2, temperature-controlled floating module 3, and optical module 4.
  • the PCR reaction chip 1 includes a chip base 101, a reaction tank 102 is provided on the chip base 101, and the surface of the reaction tank 102 is covered with a bottom film 103, forming a reaction channel with the reaction tank 102;
  • the chip base 101 is provided with a cylindrical sampling hole 104.
  • the bottom opening of the cylindrical sampling hole 104 is connected with one end of the reaction channel to form a sampling channel L1.
  • the upper end of the hole wall of the cylindrical sampling hole 104 is open.
  • the hole extends through the hole wall and communicates with the other end of the reaction channel to form an air outlet channel L2.
  • the cylindrical sampling hole 104 is closed through a threaded connection with the sampling hole cover 105.
  • the sampling hole cover 105 corresponds to the cylindrical sampling hole.
  • An extrusion head 106 is provided at position 104. As the sampling hole cover 105 rotates, the extrusion head 106 squeezes the sample into the reaction channel.
  • the reaction tank 12 has a flat waist channel structure
  • the cylindrical sampling hole 104 has an external thread cylindrical structure
  • the corresponding sampling hole cover 105 has an internal thread cover structure
  • the cylindrical sampling hole 104 has a Three-section structure, the upper part is an open truncated cone structure, the middle part is a cylindrical structure, and the bottom part is a conical structure with a hole in the center of the cone bottom;
  • the extrusion head 106 is a cylinder with a diameter interference fit in the middle part of the cylindrical sampling hole 104 structure, the extrusion head 106 is a soft sealing column, which can squeeze all the liquid in the sampling hole 104 into the reaction channel, and the inner bottom of the sampling hole cover 105 corresponds to the opening position of the cylindrical sampling hole 104
  • a sealing ring 108 is provided.
  • the chip is an integral consumable, and different freeze-dried reagents are pre-installed in the chip for different detection items.
  • the chip mounting module 2 includes a chip mounting base plate 201.
  • the mounting base plate 201 is provided with a guide slot 202, and the PCR reaction chip 1 is placed in the guide slot 202; the PCR reaction chip 1.
  • Positioning grooves 107 are provided on both sides.
  • the guide slot 202 is provided with positioning protrusions corresponding to the positioning grooves 107, and is provided with a first in-position sensor 203.
  • the chip mounting module 2 also includes a chip sliding bottom plate 204.
  • the chip sliding bottom plate 204 is provided with a chip guide rail 206 and a second in-position sensor 207.
  • the driving end is connected to the chip mounting bottom plate 201 to control the chip. It moves along the guide rail and identifies the position through the second position sensor 207.
  • the temperature-controlled floating module 3 includes a temperature control device 301.
  • the end of the temperature control device 301 is provided with a heating and cooling plate 302 and a temperature sensor 303.
  • the heating and cooling plate 302 is provided with a PCR reaction device.
  • Chip 1 corresponds to the heating and cooling carrier plate 304.
  • the heating and cooling carrier plate 304 is provided with thermal insulation pressure plates 305 on both sides, and is pressed against the temperature control device 301 through the thermal insulation pressure plates 305.
  • the temperature control device 301 is placed on the temperature control main bracket 307 through the L-shaped fixing frame 306.
  • the temperature control device 301 is connected to the L-shaped fixing frame 306 through the first temperature control guide rail 308.
  • the L-shaped fixing frame 306 is placed on the main temperature control bracket 307 through the second temperature control guide rail 310, and the second motor 311 drives the temperature control device 301 to the PCR reaction chip. 1 movement, and set the second position sensor 312.
  • the optical module 4 includes a switching device 401 and a position sensor 402; a through hole 208 is provided on the back of the chip mounting base 201 as part of the optical path for excitation light and fluorescence to pass.
  • the optical path of the optical module 4 generates excitation light from the excitation light source 403, passes through the filter 404 and lens 405, and is reflected to the chip reaction tank 102 by the secondary beam splitter 406.
  • the generated fluorescence is detected by fluorescence through the secondary beam splitter 406.
  • the device receives, where the fluorescence detection device is a photodiode 407, a camera 408 or a PMT.
  • the chip After the chip is inserted into place, it moves up and down driven by the motor so that the center of the chip coincides with the center of the heating and cooling carrier plate to prepare for subsequent compression.
  • the motor drives the temperature control module to move forward and backward in the plane, so that the heating and cooling carrier plate is close to and away from the chip.
  • the spring makes the temperature control module have a certain pressing force. Through overvoltage, the heating and cooling carrier plate is pressed tightly against the surface of the chip, making the chip The reaction chamber on the chip is in close contact with the heating and cooling slide. At the same time, the periphery of the chip is protected by an insulating pressure plate to form an insulation layer, so that the reagents in the chip reaction chamber are heated evenly and the temperature rises and falls quickly.
  • the optical module integrates and couples optical components such as lenses, filters, and secondary beam splitters to simplify the optical path and improve the stability and speed of detection.
  • the output wavelengths of the light source are 470nm, 520, 570nm, and 635nm respectively.
  • the light source is electrically coupled to produce monochromatic light, which is incident into the reaction cavity on the chip through the secondary beam splitter.
  • the fluorescence is then detected by the fluorescence detection device through the secondary beam splitter and filter.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)

Abstract

Instrument portable de PCR quantitative fluorescente ultrarapide en temps réel, comprenant : une puce pour réaction PCR (1) ; un module de montage de puce (2) utilisé pour monter la puce pour réaction PCR (1) ; un module flottant de régulation de la température (3) comprenant un dispositif de régulation de la température (301), dont une extrémité est pourvue d'une feuille de chauffage et de réfrigération (302) et d'un capteur de température (303), la feuille de chauffage et de réfrigération (302) étant pourvue d'un plateau porteur de chauffage et de réfrigération (304), deux côtés de la plaque de support de chauffage et de réfrigération (304) sont pourvus de plateaux de pression d'isolation thermique (305), et les plateaux de pression d'isolation thermique (305) sont pressés contre le module de montage de puce (2), afin que la puce pour réaction PCR (1) soit placée sur la plaque de support de chauffage et de réfrigération (304) ; et un module optique (4) comprenant un dispositif de commutation (401) et un capteur en place (402). Cet instrument de PCR est facile à transporter et à transférer ; une seule puce est utilisée, afin que l'opération soit simple, que l'étanchéité soit fiable, qu'un résultat de détection puisse être obtenu rapidement et de manière stable et que le résultat de détection soit fiable ; l'instrument de PCR peut être appliqué à une variété d'objets de détection différents, et présente une applicabilité plus large.
PCT/CN2022/083373 2022-03-23 2022-03-28 Instrument portable de pcr quantitative fluorescente ultrarapide en temps réel WO2023178710A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202220646804.9U CN217351365U (zh) 2022-03-23 2022-03-23 一种便携式超快速实时荧光定量pcr仪
CN202220646804.9 2022-03-23

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WO2023178710A1 true WO2023178710A1 (fr) 2023-09-28

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Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060246580A1 (en) * 2005-05-02 2006-11-02 Jin-Tae Kim Polymerase chain reaction module, and multiple polymerase chain reaction system including the module
CN106929408A (zh) * 2017-01-26 2017-07-07 深圳市尚维高科有限公司 便携式微流控pcr仪及基因样品荧光定量检测方法
CN106957788A (zh) * 2017-03-19 2017-07-18 北京化工大学 一种多通道实时荧光定量pcr微流控芯片系统
CN107653186A (zh) * 2017-09-15 2018-02-02 广东达元食品药品安全技术有限公司 一种便携式pcr仪
CN110551622A (zh) * 2019-09-18 2019-12-10 无锡百泰克生物技术有限公司 快速pcr反应芯片以及快速荧光定量检测仪
WO2020067742A1 (fr) * 2018-09-28 2020-04-02 주식회사 바이오메듀스 Dispositif de détection de fluorescence d'amplification en chaîne par polymérase, en temps réel
CN210945600U (zh) * 2019-09-18 2020-07-07 无锡百泰克生物技术有限公司 快速pcr反应芯片以及快速荧光定量检测仪
CN111607509A (zh) * 2019-02-22 2020-09-01 西安天隆科技有限公司 一种用于数字pcr的自动化扩增及检测装置
CN213506974U (zh) * 2020-07-31 2021-06-22 北京贝泰科技有限公司 便携式实时荧光定量pcr仪
CN214300158U (zh) * 2020-12-01 2021-09-28 无锡百泰克生物技术有限公司 一种快速荧光定量检测仪
CN113832022A (zh) * 2021-09-23 2021-12-24 吉特吉生物技术(苏州)有限公司 一种一体式核酸扩增检测设备

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060246580A1 (en) * 2005-05-02 2006-11-02 Jin-Tae Kim Polymerase chain reaction module, and multiple polymerase chain reaction system including the module
CN106929408A (zh) * 2017-01-26 2017-07-07 深圳市尚维高科有限公司 便携式微流控pcr仪及基因样品荧光定量检测方法
CN106957788A (zh) * 2017-03-19 2017-07-18 北京化工大学 一种多通道实时荧光定量pcr微流控芯片系统
CN107653186A (zh) * 2017-09-15 2018-02-02 广东达元食品药品安全技术有限公司 一种便携式pcr仪
WO2020067742A1 (fr) * 2018-09-28 2020-04-02 주식회사 바이오메듀스 Dispositif de détection de fluorescence d'amplification en chaîne par polymérase, en temps réel
CN111607509A (zh) * 2019-02-22 2020-09-01 西安天隆科技有限公司 一种用于数字pcr的自动化扩增及检测装置
CN110551622A (zh) * 2019-09-18 2019-12-10 无锡百泰克生物技术有限公司 快速pcr反应芯片以及快速荧光定量检测仪
CN210945600U (zh) * 2019-09-18 2020-07-07 无锡百泰克生物技术有限公司 快速pcr反应芯片以及快速荧光定量检测仪
CN213506974U (zh) * 2020-07-31 2021-06-22 北京贝泰科技有限公司 便携式实时荧光定量pcr仪
CN214300158U (zh) * 2020-12-01 2021-09-28 无锡百泰克生物技术有限公司 一种快速荧光定量检测仪
CN113832022A (zh) * 2021-09-23 2021-12-24 吉特吉生物技术(苏州)有限公司 一种一体式核酸扩增检测设备

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