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WO2022011706A1 - Appareil de réaction biochimique et son application - Google Patents

Appareil de réaction biochimique et son application Download PDF

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Publication number
WO2022011706A1
WO2022011706A1 PCT/CN2020/102791 CN2020102791W WO2022011706A1 WO 2022011706 A1 WO2022011706 A1 WO 2022011706A1 CN 2020102791 W CN2020102791 W CN 2020102791W WO 2022011706 A1 WO2022011706 A1 WO 2022011706A1
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WO
WIPO (PCT)
Prior art keywords
reaction
cavity
end wall
reagent
hole
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/CN2020/102791
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English (en)
Chinese (zh)
Inventor
赵静
赵霞
陈芳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
MGI Tech Co Ltd
Original Assignee
MGI Tech Co Ltd
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 MGI Tech Co Ltd filed Critical MGI Tech Co Ltd
Priority to PCT/CN2020/102791 priority Critical patent/WO2022011706A1/fr
Priority to CN202080098299.2A priority patent/CN115279498A/zh
Publication of WO2022011706A1 publication Critical patent/WO2022011706A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • 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
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present invention relates to the field of biochemical reaction devices, in particular to a biochemical reaction device capable of sealing the reaction device and preventing products from causing pollution and its application.
  • the CRISPR system is widely used in academia, mainly focusing on gene editing functions.
  • the application products developed using the CRISPR system of Cas9, Cas12, Cas13 or Cas14 are the main ones.
  • the CRISPR system using Cas12, Cas13 or Cas14 can be used for pathogen detection.
  • Cas12, Cas13 or Cas14 binds to specific gRNA to form a complex.
  • gRNA Under the guidance of gRNA, it can specifically recognize the target nucleic acid sequence (Cas12 mainly recognizes double-stranded DNA, Cas13 mainly recognizes RNA, and Cas14 mainly recognizes single-stranded DNA),
  • the target nucleic acid sequence is cleaved, and the recognition of the target nucleic acid sequence also triggers its non-specific cleavage of the surrounding single-stranded DNA or RNA.
  • the existing detection system based on CRISPR technology performs pathogen detection, it is first necessary to amplify and enrich the target DNA fragment to be detected, and then add the amplified product to the CRISPR detection system for signal detection; among them, the amplification reaction and CRISPR detection reactions were performed in separate separate test tubes (reaction systems).
  • the amplification reaction and CRISPR detection reactions were performed in separate separate test tubes (reaction systems).
  • the product molecules will be exposed outside the test tube, which will easily cause the product molecules to contaminate the working environment and cause subsequent follow-up. A false positive reaction occurred in the sample test.
  • An embodiment of the present invention provides a biochemical reaction device including a reaction vessel and a partition.
  • the reaction vessel includes a first end wall and a second end wall oppositely arranged, and a side wall connecting the first end wall and the second end wall, the first end wall and the second end wall and the side wall to form a sealed cavity.
  • the separating part is arranged on the side wall and divides the sealing cavity into a first reaction cavity and a second reaction cavity which are isolated from each other.
  • the reaction chamber is used for containing the second reaction reagent. Wherein, the partition can be opened to mix the second reactant and the first reactant.
  • the biochemical reaction device further includes a driving member, the driving member is movably arranged on the reaction vessel and includes a first end and a second end oppositely arranged, and the first end is separated from the The second end is exposed out of the reaction container, and the driving member is used to drive the partition to open.
  • the side wall is provided with a first through hole, the second end is exposed to the outside of the reaction vessel through the first through hole, and the driving member is used to pull the partition toward the The sidewall moves to tear the divider.
  • the first end wall or the second end wall is provided with a second through hole, the second end passes through the second through hole and is exposed outside the reaction vessel, and the One end of the partition part is fixedly arranged on the side wall, the other end of the partition part is movably arranged on the side wall, and the driving member is used to push the partition part toward the corresponding second end wall or The first end wall moves to open the divider.
  • the partition portion includes a protruding portion protruding toward the first end wall or the second end wall, and the protruding portion is provided with a mutually communicating accommodating cavity and a through hole, so The accommodating cavity and the through hole communicate with the first reaction cavity and the second reaction cavity together, and the through hole can make the first reaction reagent or the second reaction reagent accommodated in the accommodating cavity in the No flow out of the through holes when subjected to external pressure.
  • the biochemical reaction device further includes a driving member, the driving member is movably disposed on the reaction container and capable of pressing the first reaction reagent or the second reaction reagent accommodated in the accommodating cavity from the reaction vessel. out of the through hole.
  • the driving member includes a driving part and a partition, the driving part is movably arranged on the first end wall or the second end wall, and the partition is slidably connected to the side wall and separates the first reaction chamber or the second reaction chamber into a first part and a second part which are isolated from each other.
  • the reaction container includes an elastic part, and the elastic part can be deformed to press the first reaction reagent or the second reaction reagent accommodated in the accommodating cavity to flow out of the through hole.
  • the reaction vessel includes a first reaction part and a second reaction part, the first reaction part is provided with a first cavity, the second reaction part is provided with a second cavity, and the first reaction part is provided with a second cavity.
  • Two reaction parts are detachably mounted on the first reaction part, so that the second cavity and the first cavity together form a sealed cavity.
  • An embodiment of the present invention also provides a detection system based on CRISPR technology, comprising any of the above-mentioned biochemical reaction devices, a first reaction reagent and a second reaction reagent.
  • the first reaction reagent is accommodated in the first reaction chamber, and is used for the amplification and enrichment reaction of the sample to be detected.
  • the second reaction reagent is accommodated in the second reaction chamber, and is used to perform a CRISPR detection reaction on the amplified enriched product.
  • the biochemical reaction device provided by the present invention does not need to open the cover during use, and the entire biochemical reaction process is carried out in the sealed cavity of the reaction container, thereby avoiding the reaction product polluting the external working environment and preventing false positive reactions.
  • FIG. 1 is a schematic structural diagram of a biochemical reaction device provided in a first embodiment of the present invention.
  • FIG. 2 is a schematic structural diagram of a biochemical reaction device provided by a second embodiment of the present invention.
  • FIG. 3 is a schematic structural diagram of a biochemical reaction device provided by a third embodiment of the present invention.
  • FIG. 4 is a schematic structural diagram of a biochemical reaction device provided by a fourth embodiment of the present invention.
  • FIG. 5 is a schematic structural diagram of a biochemical reaction device according to a fifth embodiment of the present invention.
  • FIG. 1 is a schematic structural diagram of a biochemical reaction device 100 according to a first embodiment of the present invention.
  • the biochemical reaction device 100 includes a reaction vessel 10 and a partition 30 .
  • the reaction vessel 10 includes a first end wall 11 and a second end wall 13 disposed opposite to each other, and a side wall 15 connecting the first end wall 11 and the second end wall 13 .
  • the first end wall 11 , the second end wall 13 and the side wall 15 enclose a sealed cavity 16 .
  • the sealed cavity 16 is not communicated with the external working environment.
  • the partition 30 is disposed on the side wall 15 and partitions the sealed cavity 16 into a first reaction cavity 161 and a second reaction cavity 163 which are isolated from each other.
  • the first reaction chamber 161 is used to hold the first reaction reagent, and the first reaction reagent is a reactant capable of generating the first biochemical reaction; the second reaction chamber 163 is used to hold the second reaction reagent, so the The second reaction reagent is a reactant that can undergo a second biochemical reaction with the reaction product obtained from the first biochemical reaction.
  • the partition 30 can be opened to communicate with the first reaction chamber 161 and the second reaction chamber 163 , so that the reaction product in the first reaction chamber 161 and the reaction in the second reaction chamber 163 are reacted
  • the phases are mixed for a second biochemical reaction.
  • the entire biochemical reaction process is carried out in the sealed cavity 16 of the reaction vessel 10, so as to prevent the reaction product from polluting the external working environment and prevent false positive reactions.
  • the peripheral edge of the partition portion 30 is fixedly connected to the side wall 15 .
  • a first through hole 151 is formed on the side wall 15 , and the first through hole 151 is disposed adjacent to one side of the partition portion 30 .
  • the biochemical reaction device 100 further includes a driving member 50 movably disposed on the side wall 15 , and the driving member 50 is used to drive the partition portion 30 to open.
  • the driving member 50 includes a first end 51 and a second end 53 disposed opposite to each other. The first end 51 is connected to the side of the partition 30 away from the first through hole 151 , and the second end 53 passes through the first through hole 151 and is exposed to the outside of the side wall 15 .
  • the dividing portion 30 moves toward the side wall 15 along with the driving member 50 to tear the dividing portion 30, thereby opening the dividing portion 30, so that the first The first reaction reagent in a reaction chamber 161 and the second reaction reagent in the second reaction chamber 163 can be mixed.
  • the partition portion 30 is a film
  • the driving member 50 is a pull rod.
  • a sealing member (not shown) is further provided between the first through hole 151 and the driving member 50 to seal the sealing cavity 16 .
  • the reaction vessel 10 includes a first reaction part 12 and a second reaction part 14 .
  • the first reaction part 12 includes a bottom wall and a side wall surrounding the bottom wall, wherein the bottom wall and the side wall enclose a first cavity.
  • the second reaction part 14 includes a bottom wall and a side wall surrounding the bottom wall, wherein the bottom wall and the side wall enclose a second cavity.
  • the side wall of the second reaction part 14 is detachably mounted on the side wall of the first reaction part 12 , so that the first cavity and the second cavity together form the sealed cavity 16 .
  • the first reaction part 12 is a test tube
  • the second reaction part 14 is a tube cover.
  • the side wall of the second reaction part 14 is screwed to the side wall of the first reaction part 12 .
  • the second reaction part 14 can also be detachably connected to the first reaction part 12 by other connection means, such as interference fit.
  • a sealing member (not shown) may be further provided between the side wall of the second reaction part 14 and the side wall of the first reaction part 12 for sealing the sealing cavity 16 .
  • the periphery of the partition portion 30 is fixedly connected to the side wall of the second reaction portion 14 ; the second reaction cavity 163 is located in the second cavity of the second reaction portion 14 , so The first reaction cavity 161 is located in the first cavity of the first reaction part 12 and part of the second cavity of the second reaction part 14; the driving member 50 and the first through hole 151 are located in the The separation part 30 is at a side away from the first reaction part 12 .
  • the driving member 50 is pulled, the second reaction reagent in the second reaction chamber 163 flows into the first reaction chamber 161 and is mixed with the first reaction reagent.
  • the first reaction chamber 161 can be used to hold the second reaction reagent
  • the second reaction chamber 163 can be used to hold the first reaction reagent.
  • the driving member 50 is pulled, the The first reactant may flow into the second reactant for mixing.
  • the biochemical reaction device 100 is suitable for a biochemical reaction system with two or more reaction steps, and the reaction product obtained in one of the reaction steps needs to be mixed with the reactants of the subsequent steps to carry out the reaction; it is also applicable to the need to open the cover for additional A biochemical reaction system for adding reagents.
  • the biochemical reaction device 100 can be applied to an RNA reverse transcription system or a detection system based on CRISPR technology. In this embodiment, the biochemical reaction device 100 is applied to a detection system based on CRISPR technology for example.
  • the detection system based on CRISPR technology includes the biochemical reaction device 100, a first reaction reagent and a second reaction reagent.
  • the first reaction reagent is preset in the first reaction chamber 161, and is used for amplification and enrichment reaction of the DNA fragment to be detected.
  • the first reaction reagent can be any existing reagent that can amplify and enrich DNA fragments.
  • the second reaction reagent is pre-placed in the second reaction chamber 163 for performing the CRISPR detection reaction on the amplified enriched product.
  • the second reaction reagent can be any reagent currently required for CRISPR detection reaction.
  • the driving member 50 pulls the partition 30 to move toward the side wall 15 , so that the partition 30 is torn along the direction perpendicular to the side wall 15 , so that the second reaction reagent in the second reaction chamber 163 flows into the first reaction chamber 161 to be mixed with the amplification enrichment product.
  • the second reaction reagent by adding the second reaction reagent to the amplification and enrichment product, the number of cycles of amplification and enrichment can be greatly shortened, thereby shortening the overall detection time.
  • FIG. 2 is a schematic structural diagram of a biochemical reaction device 100 according to a second embodiment of the present invention.
  • the difference between the biochemical reaction apparatus 100 shown in this embodiment and the biochemical reaction apparatus 100 shown in the first embodiment is that the driving member 50 is movably arranged on the second end wall 13 , and one end of the partition 30 is movably disposed on the second end wall 13 .
  • the movable part is arranged on the side wall 15 , so that the driving member 50 pushes the partition part 30 to open toward the first end wall 11 , so as to change the opening method of the partition part 30 .
  • the driving member 50 may also be movably disposed on the first end wall 11 , and the driving member 50 can push one end of the partition portion 30 toward the second end Wall 13 opens.
  • the second end wall 13 defines a second through hole (not shown), and the second end 53 of the driving member 50 protrudes out of the reaction vessel 10 through the second through hole .
  • the driving member 50 is a button assembly.
  • One end of the partition part 30 is fixedly connected to the side wall 15 , and the other end of the partition part 30 is movably connected to the side wall 15 , so that the driving member 50 pushes the partition part 30 toward the first
  • one end of the partition 30 opens toward the first end wall 11 , so that the reaction product in the first reaction chamber 161 and the reactant in the second reaction chamber 163 are mixed , for the second biochemical reaction to take place.
  • the driver 50 Push the partition 30 to move toward the first end wall 11, so that the end of the partition 30 that is movably connected to the side wall 15 is opened toward the first end wall 11, so that the second The second reaction reagent in the reaction chamber 163 flows into the first reaction chamber 161 to be mixed with the amplified enriched product.
  • FIG. 3 is a schematic structural diagram of a biochemical reaction device 100 according to a third embodiment of the present invention.
  • the difference between the biochemical reaction apparatus 100 of this embodiment and the biochemical reaction apparatus 100 shown in the second embodiment is that the partition 30 shown is not opened by the driving of the driving member 50 , but is opened by the partition The pressure on the side of the part 30 changes and is opened.
  • the reaction vessel 10 includes an elastic part, and the elastic part can be deformed to increase the pressure on one side of the partition part 30 , thereby forcing the end of the partition part 30 movably connected to the side wall 15 to face
  • the first end wall 11 is open.
  • the elastic portion may be located at at least one of the side wall 15 , the first end wall 11 and the second end wall 13 of the reaction vessel 10 .
  • the second reaction part 14 is made of elastic material, and the second reaction part 14 acts as an elastic part of the reaction container 10 as a whole. When the second reaction part 14 is compressed and deformed, the pressure of the partition part 30 on the side of the second reaction chamber 163 is increased, thereby pressing the partition part 30 to be movably connected to the side wall 15 .
  • the first reaction part 12 can be made of an elastic material, and the first reaction part 12 acts as an elastic part of the reaction container 10 as a whole.
  • the first reaction part 12 When compressed and deformed, the pressure of the partition 30 on the side of the first reaction chamber 161 is increased, so that the end of the partition 30 that is movably connected to the side wall 15 is pressed toward the second end wall 13 Open.
  • the biochemical reaction device 100 of this embodiment When the biochemical reaction device 100 of this embodiment is applied to a detection system based on CRISPR technology, after the first reaction reagent in the first reaction chamber 161 and the DNA to be detected undergo an amplification and enrichment reaction, the first reaction chamber 161 is compressed. A reaction part 12 deforms it, thereby increasing the pressure of the partition part 30 on the side of the first reaction chamber 161 , thereby forcing the end of the partition part 30 movably connected to the side wall 15 to face the The second end wall 13 is opened, so that the second reaction reagent located in the second reaction chamber 163 flows into the first reaction chamber 161 to be mixed with the amplification enrichment product.
  • FIG. 4 is a schematic structural diagram of a biochemical reaction device 100 according to a fourth embodiment of the present invention.
  • the difference between the biochemical reaction apparatus 100 of this embodiment and the biochemical reaction apparatus 100 shown in the second embodiment is that the structure of the partition part 30 is different, and the connection relationship between the driving member 50 and the partition part 30 is different .
  • the partition portion 30 includes a protruding portion 32 formed to protrude toward the first end wall 11 .
  • a receiving cavity 321 and a through hole 323 are sequentially opened on the protruding portion 32 along a direction perpendicular to the first end wall 11 , and an outlet (not shown) is formed at the end of the through hole 323 away from the receiving cavity 321 .
  • the through hole 323 communicates with the accommodating cavity 321 , and the accommodating cavity 321 and the through hole 323 communicate with the second reaction cavity 163 and the first reaction cavity 161 together.
  • the through hole 323 can prevent the second reaction reagent accommodated in the accommodating cavity 321 from flowing out of the through hole 323 when the external pressure is not applied by the driving member 50 .
  • the partition portion 30 when the second reaction reagent in the accommodating cavity 321 cannot flow out from the through hole 323 , the partition portion 30 is in an unopened state; When the second reaction reagent can flow out from the through hole 323, the partition 30 is in an open state.
  • the accommodating cavity 321 is substantially in the shape of a frusto-inverted cone
  • the through hole 323 is substantially cylindrical
  • the diameter of the through hole 323 is smaller than or equal to the minimum diameter of the accommodating cavity 321 .
  • the drive member 50 includes a drive portion 52 and a diaphragm 54 that are connected to each other.
  • the driving portion 52 is movably installed in the second through hole and partially extends out of the reaction vessel 10 .
  • the partition plate 54 is slidably connected to the side wall 15 and divides the second reaction chamber 163 into a first part and a second part which are isolated from each other. The second part is used for containing the second reaction reagent.
  • the protruding portion 32 may also be formed to protrude toward the second end wall 13 , and the driving portion 52 is movably disposed on the first end wall 11 and can Pushing the partition plate 54 presses the first reaction reagent in the first reaction chamber 161 to flow out into the second reaction chamber 163 through the through hole 323 to be mixed with the second reaction reagent.
  • the driving part will 52 pushes the partition plate 54 to press the second reaction reagent located in the second reaction chamber 163 to flow out into the first reaction chamber 161 through the through hole 323 to be mixed with the enriched amplification product.
  • FIG. 5 is a schematic structural diagram of a biochemical reaction device 100 according to a fifth embodiment of the present invention.
  • the difference between the biochemical reaction apparatus 100 of this embodiment and the biochemical reaction apparatus 100 provided in the fourth embodiment is that the partition 30 shown is not opened by the driving of the driving member 50 , but is opened by the partition 30 side of the pressure changes and is opened.
  • the reaction container 10 includes an elastic part, and the elastic part can be deformed to increase the pressure on the side of the second reaction reagent accommodated in the accommodating cavity 321 , thereby compressing the second reaction reagent from The through hole 323 flows out into the first reaction chamber 161 to be mixed with the first reaction reagent.
  • the second reaction part 14 is made of elastic material. When the second reaction part 14 is compressed and deformed, the pressure on the side of the second reaction reagent accommodated in the accommodating cavity 321 is reduced by increase, thereby forcing the second reaction reagent to flow out from the through hole 323 to open the partition 30 .
  • the protruding portion 32 may also be formed to protrude toward the second end wall 13 , and the first reaction portion 12 may be made of an elastic material.
  • the portion 12 When the portion 12 is compressed and deformed, the pressure on the side of the first reaction reagent accommodated in the accommodating cavity 321 is increased, thereby forcing the first reaction reagent to flow out of the through hole 323 to open the Separator 30 .
  • the first reaction part 12 and the second reaction part 14 are both test tubes.
  • the biochemical reaction device 100 of this embodiment When the biochemical reaction device 100 of this embodiment is applied to a detection system based on CRISPR technology, after the first reaction reagent in the first reaction chamber 161 and the DNA to be detected undergo an amplification and enrichment reaction, the first reaction chamber 161 is compressed. The second reaction part 14 is deformed to press the second reaction reagent in the second reaction chamber 163 to flow out into the first reaction chamber 161 through the through hole 323 to be mixed with the enriched product.

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Abstract

Un appareil de réaction biochimique (100), comprenant un récipient de réaction (10) et une partie de séparation (30). Le récipient de réaction (10) comprend une première paroi d'extrémité (11) et une seconde paroi d'extrémité (13) qui sont disposées à l'opposé l'une de l'autre, et une paroi latérale (15) reliant la première paroi d'extrémité (11) et la deuxième paroi d'extrémité (13) ; la première paroi d'extrémité (11), la deuxième paroi d'extrémité (13), et la paroi latérale (15) définissent une cavité étanche (16). La partie de séparation (30) est disposée sur la paroi latérale (15) et sépare la cavité étanche (16) dans une première chambre de réaction (161) et une seconde chambre de réaction (163) ; la première chambre de réaction (161) est utilisée pour contenir un premier réactif de réaction, et la seconde chambre de réaction (163) est utilisée pour contenir un second réactif de réaction. La partie de séparation peut être ouverte pour mélanger le second réactif de réaction avec le premier réactif de réaction. L'invention concerne également un système de détection basé sur la technologie CRISPR comprenant l'appareil de réaction biochimique (100).
PCT/CN2020/102791 2020-07-17 2020-07-17 Appareil de réaction biochimique et son application Ceased WO2022011706A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2020/102791 WO2022011706A1 (fr) 2020-07-17 2020-07-17 Appareil de réaction biochimique et son application
CN202080098299.2A CN115279498A (zh) 2020-07-17 2020-07-17 生化反应装置及其应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/102791 WO2022011706A1 (fr) 2020-07-17 2020-07-17 Appareil de réaction biochimique et son application

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WO2022011706A1 true WO2022011706A1 (fr) 2022-01-20

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CN205396909U (zh) * 2015-12-05 2016-07-27 重庆利尔达科技开发有限公司 一种双组份可施胶薄膜袋
CN107523487A (zh) * 2017-09-18 2017-12-29 星源智（珠海）生物科技有限公司 一种集成化管状反应装置
CN110304325A (zh) * 2019-07-03 2019-10-08 广东米大叔农业科技有限公司 一种可盛放两种口味饮料的易拉罐

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CN117504797B (zh) * 2023-12-29 2024-05-14 常州新一产生命科技有限公司 合成柱结构及生化反应设备

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