WO2022011706A1 - 生化反应装置及其应用 - Google Patents

生化反应装置及其应用 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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Prior art keywords
reaction
cavity
end wall
reagent
hole
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English (en)
French (fr)
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赵静
赵霞
陈芳
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MGI Tech Co Ltd
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MGI Tech Co Ltd
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Priority to CN202080098299.2A priority Critical patent/CN115279498A/zh
Priority to PCT/CN2020/102791 priority patent/WO2022011706A1/zh
Publication of WO2022011706A1 publication Critical patent/WO2022011706A1/zh
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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

一种生化反应装置（100），包括反应容器（10）和分隔部（30）。反应容器（10）包括相对设置的第一端壁（11）和第二端壁（13）、以及连接第一端壁（11）和第二端壁（13）的侧壁（15），第一端壁（11）、第二端壁（13）和侧壁（15）围合成一密封腔（16）。分隔部（30）设置于侧壁（15）上并将密封腔（16）分隔为相互隔离的第一反应腔（161）和第二反应腔（163），第一反应腔（161）用于盛装第一反应试剂，第二反应腔（163）用于盛装第二反应试剂。其中，分隔部（30）能够被打开以将第二反应试剂和第一反应试剂进行混合。还提供包括生化反应装置（100）的基于CRISPR技术的检测系统。

Description

生化反应装置及其应用 技术领域
本发明涉及生物化学反应装置领域，尤其涉及一种能够封闭反应装置、防止产物造成污染的生化反应装置及其应用。
背景技术
CRISPR系统在学术界的应用甚为广泛，主要集中在基因编辑功能，近年来随着该技术的不断发展以及新型CRISPR体系的开发，相继涌现出利用该技术的应用型产品。目前，以利用Cas9、Cas12、Cas13或Cas14的CRISPR体系开发的应用产品为主。其中，利用Cas12、Cas13或Cas14的CRISPR体系可用于病原检测。检测时，Cas12、Cas13或Cas14结合特异性gRNA形成复合体，在gRNA的引导下，可特异性识别目标核酸序列(Cas12主要识别双链DNA，Cas13主要识别RNA，Cas14主要识别单链DNA)，并对目标核酸序列进行切割，对目标核酸序列的识别同时也触发其对周围的单链DNA或RNA的非特异性切割。
现有的基于CRISPR技术的检测系统在进行病原检测时，首先需将待检测目标DNA片段进行扩增富集，再将扩增产物加入到CRISPR检测系统中进行信号检测；其中，扩增反应和CRISPR检测反应分别在单独的分离的试管(反应系统)中进行。使用单独的分离的试管进行多步反应以进行检测时，在反应产物从一个试管转移到另一个试管的过程中，产物分子会暴露在试管外，容易使产物分子对工作环境造成污染，导致后续样品检测出现假阳性反应。
发明内容
鉴于上述状况，有必要提供一种能够避免产物产生污染的生化反应装置及其应用。
本发明一实施方式提供一种生化反应装置，包括反应容器和分隔部。所述反应容器包括相对设置的第一端壁和第二端壁、以及连接所述第一端壁和所述第二端壁的侧壁，所述第一端壁、所述第二端壁和所述侧壁围合成一密封腔。所述分隔部设置于所述侧壁上并将所述密封腔分隔为相互隔离的第一反应腔和第二反应腔，所述第一反应腔用于盛装第一反应试剂，所述第二反应腔用于盛装第二反应试剂。其中，所述分隔部能够被打开以将所述第二反应试剂和所述第一反应试剂进行混合。
一种实施方式中，所述生化反应装置还包括驱动件，所述驱动件活动设置于所述反应容器上并包括相对设置的第一端和第二端，所述第一端与所述分隔部相连接，所述第二端露出于所述反应容器外，所述驱动件用于驱动所述分隔部打开。
一种实施方式中，所述侧壁开设有第一通孔，所述第二端穿过所述第一通孔露出于所述反应容器外，所述驱动件用于拉动所述分隔部朝向所述侧壁运动以撕裂所述分隔部。
一种实施方式中，所述第一端壁或所述第二端壁上开设有第二通孔，所述第二端穿过所述第二通孔露出于所述反应容器外，所述分隔部的一端固定设置于所述侧壁上，所述分隔部的另一端活动设置于所述侧壁上，所述驱动件用于推动所述分隔部朝向相应的所述第二端壁或所述第一端壁运动以打开所述分隔部。
一种实施方式中，所述分隔部包括朝向所述第一端壁或所述第二 端壁凸伸形成的凸伸部，所述凸伸部开设有相互连通的容纳腔和通孔，所述容纳腔和所述通孔共同连通所述第一反应腔和所述第二反应腔，所述通孔能够使容置于所述容纳腔中的第一反应试剂或第二反应试剂在未受到外部压力时不会从所述通孔中流出。
一种实施方式中，所述生化反应装置还包括驱动件，所述驱动件活动设置于所述反应容器上并能够压迫容置于所述容纳腔中的第一反应试剂或第二反应试剂从所述通孔中流出。
一种实施方式中，所述驱动件包括驱动部和隔板，所述驱动部活动设置于所述第一端壁或所述第二端壁上，所述隔板滑动连接于所述侧壁上并将所述第一反应腔或所述第二反应腔分隔为相互隔离的第一部分和第二部分。
一种实施方式中，所述反应容器包括弹性部分，所述弹性部分能够发生形变以压迫容置于所述容纳腔中的第一反应试剂或第二反应试剂从所述通孔中流出。
一种实施方式中，所述反应容器包括第一反应部和第二反应部，所述第一反应部开设有第一腔体，所述第二反应部开设有第二腔体，所述第二反应部可拆卸地装设于所述第一反应部上使所述第二腔体和所述第一腔体共同形成一密封腔。
本发明一实施方式还提供一种基于CRISPR技术的检测系统，包括上述任一种生化反应装置、第一反应试剂和第二反应试剂。所述第一反应试剂收容于所述第一反应腔中，用于使待检测样品发生扩增富集反应。所述第二反应试剂收容于所述第二反应腔中，用于使扩增富集产物进行CRISPR检测反应。
本发明提供的生化反应装置，在使用时，无需进行开盖操作，且整个生化反应过程都在反应容器的密封腔中进行，从而避免反应产物 对外界工作环境造成污染，防止假阳性反应。
附图说明
图1为本发明第一实施方式提供的生化反应装置的结构示意图。
图2为本发明第二实施方式提供的生化反应装置的结构示意图。
图3为本发明第三实施方式提供的生化反应装置的结构示意图。
图4为本发明第四实施方式提供的生化反应装置的结构示意图。
图5为本发明第五实施方式提供的生化反应装置的结构示意图。
主要元件符号说明
生化反应装置                 100
反应容器                     10
分隔部                       30
第一端壁                     11
第二端壁                     13
侧壁                         15
密封腔                       16
第一反应腔                   161
第二反应腔                   163
第一通孔                     151
驱动件                       50
第一端                       51
第二端                       53
第一反应部                   12
第二反应部                   14
凸伸部                       32
容纳腔                       321
通孔                         323
驱动部                       52
隔板                         54
如下具体实施方式将结合上述附图进一步说明本发明。
具体实施方式
在下面的描述中阐述了很多具体细节以便于充分理解本发明，但是本发明还可以采用其他不同于在此描述的其它方式来实施，本领域技术人员可以在不违背本发明内涵的情况下做类似应用，因此本发明不受下面公开的具体实施例的限制。
请参阅图1，图1为本发明第一实施方式提供的生化反应装置100的结构示意图。所述生化反应装置100包括反应容器10以及分隔部30。所述反应容器10包括相对设置的第一端壁11和第二端壁13、以及连接所述第一端壁11和所述第二端壁13的侧壁15。所述第一端壁11、所述第二端壁13和所述侧壁15围合成一密封腔16。所述密封腔16与外界工作环境不相通。所述分隔部30设置于所述侧壁15上，并将所述密封腔16分隔为相互隔离的第一反应腔161和第二反应腔163。所述第一反应腔161用于盛装第一反应试剂，所述第一反应试剂为能够发生第一次生化反应的反应物；所述第二反应腔163用于盛放第二反应试剂，所述第二反应试剂为能够和第一次生化反应所得反应产物发生第二次生化反应的反应物。所述分隔部30能够被打开以连通所述第一反应腔161和所述第二反应腔163，从而使所述第一反应腔161中的反应产物和所述第二反应腔163中的反应物相混 合，以发生第二次生化反应。整个生化反应过程都在反应容器10的密封腔16中进行，从而避免反应产物对外界工作环境造成污染，防止假阳性反应。
所述分隔部30的周缘固定连接于所述侧壁15上。所述侧壁15上开设有第一通孔151，所述第一通孔151邻近所述分隔部30的一侧设置。所述生化反应装置100还包括活动设置于所述侧壁15上的驱动件50，所述驱动件50用于驱动所述分隔部30打开。所述驱动件50包括相对设置的第一端51和第二端53。所述第一端51与所述分隔部30远离所述第一通孔151的一侧相连接，所述第二端53穿过所述第一通孔151露出于所述侧壁15外。当拉动所述驱动件50时，所述分隔部30随同所述驱动件50朝向所述侧壁15运动，以撕裂所述分隔部30，进而使所述分隔部30打开，使得所述第一反应腔161的第一反应试剂和所述第二反应腔163中的第二反应试剂反应试剂能够混合。本实施方式中，所述分隔部30为薄膜，所述驱动件50为拉杆。
可选的，所述第一通孔151和所述驱动件50之间还设有密封件(图未示)，以密封所述密封腔16。
所述反应容器10包括第一反应部12和第二反应部14。所述第一反应部12包括底壁和环绕所述底壁设置的侧壁，其中所述底壁和所述侧壁合围形成第一腔体。所述第二反应部14包括底壁和环绕所述底壁设置的侧壁，其中所述底壁和所述侧壁合围形成第二腔体。所述第二反应部14的侧壁可拆卸地装设于所述第一反应部12的侧壁上，使所述第一腔体和所述第二腔体共同形成所述密封腔16。所述第一反应部12的侧壁和所述第二反应部14的侧壁共同构成所述反应容器10的所述侧壁15，所述第一反应部12的底壁作为所述反应容 器10的所述第一端壁11，所述第二反应部14的底壁作为所述反应容器的所述第二端壁13。本实施方式中，所述第一反应部12为试管，所述第二反应部14为管盖。
本实施方式中，所述第二反应部14的侧壁螺接于所述第一反应部12的侧壁上。在其他实施方式中，所述第二反应部14还可通过其他连接方式与所述第一反应部12可拆卸地连接，例如通过过盈配合。
可选的，所述第二反应部14的侧壁和所述第一反应部12的侧壁之间还可设置有密封件(图未示)，用于密封所述密封腔16。
本实施方式中，所述分隔部30的周缘固定连接于所述第二反应部14的侧壁上；所述第二反应腔163位于所述第二反应部14的第二腔体中，所述第一反应腔161位于所述第一反应部12的第一腔体和所述第二反应部14的部分第二腔体中；所述驱动件50和所述第一通孔151位于所述分隔部30远离所述第一反应部12的一侧。当拉动所述驱动件50时，位于所述第二反应腔163中的第二反应试剂流入所述第一反应腔161中，与所述第一反应试剂相混合。可以理解的是，在其他实施方式中，所述第一反应腔161可用于盛装第二反应试剂，所述第二反应腔163可用于盛装第一反应试剂，当拉动所述驱动件50时，可使所述第一反应试剂流入所述第二反应试剂中进行混合。
所述生化反应装置100适用于具有两步及以上反应步骤，且需将其中一反应步骤所得的反应产物和后续步骤的反应物混合以进行反应的生化反应系统；还可适用于需要开盖额外添加试剂的生化反应系统。所述生化反应装置100可应用到RNA反转录系统或基于CRISPR技术的检测系统中。本实施方式中，将所述生化反应装置100应用到基于CRISPR技术的检测系统中进行示例。
所述基于CRISPR技术的检测系统包括所述生化反应装置100、 第一反应试剂以及第二反应试剂。所述第一反应试剂预置于所述第一反应腔161中，用于使待检测DNA片段发生扩增富集反应。所述第一反应试剂可采用现有可对DNA片段进行扩增富集的任意试剂。所述第二反应试剂预置于所述第二反应腔163中，用于使扩增富集产物进行CRISPR检测反应。所述第二反应试剂可采用现有进行CRISPR检测反应所需的任何试剂。
在所述第一反应腔161中的第一反应试剂和待检测DNA发生扩增富集反应后，所述驱动件50拉动所述分隔部30朝向所述侧壁15运动，使得所述分隔部30沿垂直所述侧壁15的方向被撕裂，进而使位于所述第二反应腔163中的第二反应试剂流入所述第一反应腔161中与扩增富集产物混合。本实施方式中，通过将第二反应试剂加入扩增富集产物中，可极大缩短扩增富集循环次数，从而可缩短整体检测时间。
请参阅图2，图2为本发明第二实施方式提供的生化反应装置100的结构示意图。本实施方式所示生化反应装置100与第一实施方式所示生化反应装置100的不同之处在于，所述驱动件50活动设置于所述第二端壁13上，所述分隔部30的一端活动设置于所述侧壁15上，使得所述驱动件50推动所述分隔部30朝向所述第一端壁11打开，以改变所述分隔部30的打开方式。可以理解的是，在其他实施方式中，所述驱动件50还可活动设置于所述第一端壁11上，所述驱动件50能够推动所述分隔部30的一端朝向所述第二端壁13打开。
本实施方式中，所述第二端壁13开设有第二通孔(图未示)，所述驱动件50的第二端53穿过所述第二通孔伸出所述反应容器10外。本实施方式中，所述驱动件50为按键组件。所述分隔部30的一端与所述侧壁15固定连接，所述分隔部30的另一端与所述侧壁15 活动连接，使得所述驱动件50推动所述分隔部30朝向所述第一端壁11运动时，所述分隔部30的一端朝向所述第一端壁11打开，从而使所述第一反应腔161中的反应产物和所述第二反应腔163中的反应物相混合，以发生第二次生化反应。
当将本实施方式的生化反应装置100应用于基于CRISPR技术的检测系统时，在所述第一反应腔161中的第一反应试剂和待检测DNA发生扩增富集反应后，所述驱动件50推动所述分隔部30朝向所述第一端壁11运动，使得所述分隔部30与所述侧壁15活动连接的一端朝向所述第一端壁11打开，进而使位于所述第二反应腔163中的第二反应试剂流入所述第一反应腔161中与扩增富集产物混合。
请参阅图3，图3为本发明第三实施方式提供的生化反应装置100的结构示意图。本实施方式的生化反应装置100与第二实施方式所示的生化反应装置100的不同之处在于，所示分隔部30并非通过所述驱动件50的驱动而被打开，而是通过所述分隔部30一侧的压强变化而被打开。
具体的，所述反应容器10包括弹性部分，所述弹性部分能够发生形变以增大所述分隔部30一侧的压强，从而压迫所述分隔部30与所述侧壁15活动连接的一端朝向所述第一端壁11打开。所述弹性部分可位于所述反应容器10的侧壁15、第一端壁11和第二端壁13中至少一者处。本实施方式中，所述第二反应部14由弹性材料制成，所述第二反应部14整体作为所述反应容器10的弹性部分。当所述第二反应部14被压缩变形时，所述分隔部30位于所述第二反应腔163一侧的压强被增大，从而压迫所述分隔部30与所述侧壁15活动连接的一端朝向所述第一端壁11打开。可以理解的是，在其他实施方式中，所述第一反应部12可由弹性材料制成，所述第一反应部12整体 作为所述反应容器10的弹性部分，当所述第一反应部12被压缩变形时，所述分隔部30位于所述第一反应腔161一侧的压强被增大，从而压迫所述分隔部30与所述侧壁15活动连接的一端朝向所述第二端壁13打开。
当将本实施方式的生化反应装置100应用于基于CRISPR技术的检测系统时，在所述第一反应腔161中的第一反应试剂和待检测DNA发生扩增富集反应后，压缩所述第一反应部12使其发生形变，进而增大所述分隔部30位于所述第一反应腔161一侧的压强，从而压迫所述分隔部30与所述侧壁15活动连接的一端朝向所述第二端壁13打开，使位于所述第二反应腔163中的第二反应试剂流入所述第一反应腔161中与扩增富集产物混合。
请参阅图4，图4为本发明第四实施方式提供的生化反应装置100的结构示意图。本实施方式的生化反应装置100与第二实施方式所示的生化反应装置100的不同之处在于，所述分隔部30的结构不同，所述驱动件50与所述分隔部30的连接关系不同。
所述分隔部30包括朝向所述第一端壁11凸伸形成的凸伸部32。所述凸伸部32沿垂直于所述第一端壁11的方向依次开设有容纳腔321和通孔323，所述通孔323背离所述容纳腔321的一端设有出口(图未示)。所述通孔323与所述容纳腔321相连通，所述容纳腔321和所述通孔323共同连通所述第二反应腔163和所述第一反应腔161。所述通孔323能够使容纳于所述容纳腔321中的第二反应试剂在未受到所述驱动件50施加的外部压力时不会从所述通孔323中流出。需要说明的是，本发明中，当所述容纳腔321中的第二反应试剂不能够从所述通孔323中流出时，所述分隔部30处于未打开状态；当所述容纳腔321中的第二反应试剂能够从所述通孔323中流出时， 所述分隔部30处于打开状态。
本实施方式中，所述容纳腔321大致为截头倒锥形，所述通孔323大致为圆柱状，所述通孔323的直径小于或等于所述容纳腔321的最小直径。本实施方式中，当所述容纳腔321中的第二反应试剂仅受到所述第一反应腔161和所述第二反应腔163中的空气的压强的作用时，而未受到外部压力时，其在空气压强的作用下，不会从所述通孔323中流出。也就是说，当容纳于所述容纳腔321中的第二反应试剂在未受到所述驱动件50施加的外部压力时，所述第一反应腔161中的空气对所述第二反应试剂施加的压强应大于所述第二反应腔163中的空气对所述第二反应试剂施加的压强，从而使所述第二反应试剂不会从所述通孔323中流出。
所述驱动件50包括相互连接的驱动部52和隔板54。所述驱动部52活动装设于所述第二通孔中并部分伸出于所述反应容器10外。所述隔板54滑动连接于所述侧壁15上，并将所述第二反应腔163分隔为相互隔离的第一部分和第二部分。所述第二部分用于盛装所述第二反应试剂。当所述驱动部52推动所述隔板54沿着所述侧壁15滑动时，所述第二反应腔163中的第一部分的体积增大，所述第二反应腔163中的第二部分的体积缩小，使得所述第二反应试剂在外部压力的作用下从所述通孔323中流出至所述第一反应腔161中与所述第一反应试剂混合。
可以理解的是，在其他实施方式中，所述凸伸部32还可朝向所述第二端壁13凸伸形成，所述驱动部52活动设置于所述第一端壁11上，并能够推动所述隔板54压迫位于所述第一反应腔161中的第一反应试剂通过所述通孔323流出至所述第二反应腔163中与所述第二反应试剂混合。
当将本实施方式的生化反应装置100应用于基于CRISPR技术的检测系统时，在所述第一反应腔161中的第一反应试剂和待检测DNA发生扩增富集反应后，所述驱动部52推动所述隔板54压迫位于所述第二反应腔163中的第二反应试剂通过所述通孔323流出至所述第一反应腔161中与扩增富集产物混合。
请参阅图5，图5为本发明第五实施方式提供的生化反应装置100的结构示意图。本实施方式的生化反应装置100与第四实施方式提供的生化反应装置100的不同之处在于，所示分隔部30并非通过所述驱动件50的驱动而被打开，而是通过所述分隔部30一侧的压强变化而被打开。
具体的，所述反应容器10包括弹性部分，所述弹性部分能够发生形变以增大容置于所述容纳腔321中的第二反应试剂一侧的压强，从而压迫所述第二反应试剂从所述通孔323中流出至所述第一反应腔161中与所述第一反应试剂混合。本实施方式中，所述第二反应部14由弹性材料制成，当所述第二反应部14被压缩变形时，容置于所述容纳腔321中的第二反应试剂一侧的压强被增大，从而压迫所述第二反应试剂从所述通孔323中流出，以打开所述分隔部30。
可以理解的是，在其他实施方式中，所述凸伸部32还可朝向所述第二端壁13凸伸形成，所述第一反应部12可由弹性材料制成，当所述第一反应部12被压缩变形时，容置于所述容纳腔321中的第一反应试剂一侧的压强被增大，从而压迫所述第一反应试剂从所述通孔323中流出，以打开所述分隔部30。本实施方式中，所述第一反应部12和所述第二反应部14均为试管。
当将本实施方式的生化反应装置100应用于基于CRISPR技术的检测系统时，在所述第一反应腔161中的第一反应试剂和待检测DNA 发生扩增富集反应后，压缩所述第二反应部14使其发生形变进而压迫位于所述第二反应腔163中的第二反应试剂通过所述通孔323流出至所述第一反应腔161中与扩增富集产物混合。
以上实施方式仅用以说明本发明的技术方案而非限制，对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的，本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此，本发明将不会被限制于本文所示的这些实施例，而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。

Claims (10)

  1. 一种生化反应装置，包括：
    反应容器，包括相对设置的第一端壁和第二端壁、以及连接所述第一端壁和所述第二端壁的侧壁，所述第一端壁、所述第二端壁和所述侧壁围合成一密封腔；以及
    分隔部，设置于所述侧壁上并将所述密封腔分隔为相互隔离的第一反应腔和第二反应腔，所述第一反应腔用于盛装第一反应试剂，所述第二反应腔用于盛装第二反应试剂；
    其中，所述分隔部能够被打开以将所述第二反应试剂和所述第一反应试剂进行混合。
  2. 如权利要求1所述的生化反应装置，其特征在于，所述生化反应装置还包括驱动件，所述驱动件活动设置于所述反应容器上并包括相对设置的第一端和第二端，所述第一端与所述分隔部相连接，所述第二端露出于所述反应容器外，所述驱动件用于驱动所述分隔部打开。
  3. 如权利要求2所述的生化反应装置，其特征在于，所述侧壁开设有第一通孔，所述第二端穿过所述第一通孔露出于所述反应容器外，所述驱动件用于拉动所述分隔部朝向所述侧壁运动以撕裂所述分隔部。
  4. 如权利要求2所述的生化反应装置，其特征在于，所述第一端壁或所述第二端壁上开设有第二通孔，所述第二端穿过所述第二通孔露出于所述反应容器外，所述分隔部的一端固定设置于所述侧壁上，所述分隔部的另一端活动设置于所述侧壁上，所述驱动件用于推动所述分隔部朝向相应的所述第二端壁或所述第一端壁运动以打开所述分隔部。
  5. 如权利要求1所述的生化反应装置，其特征在于，所述分隔部包括朝向所述第一端壁或所述第二端壁凸伸形成的凸伸部，所述凸伸部开设有相互连通的容纳腔和通孔，所述容纳腔和所述通孔共同连通所述第一反应腔和所述第二反应腔，所述通孔能够使容置于所述容纳腔中的第一反应试剂或第二反应试剂在未受到外部压力时不会从所述通孔中流出。
  6. 如权利要求5所述的生化反应装置，其特征在于，所述生化反应装置还包括驱动件，所述驱动件活动设置于所述反应容器上并能够压迫容置于所述容纳腔中的第一反应试剂或第二反应试剂从所述通孔中流出。
  7. 如权利要求6所述的生化反应装置，其特征在于，所述驱动件包括驱动部和隔板，所述驱动部活动设置于所述第一端壁或所述第二端壁上，所述隔板滑动连接于所述侧壁上并将所述第一反应腔或所述第二反应腔分隔为相互隔离的第一部分和第二部分。
  8. 如权利要求5所述的生化反应装置，其特征在于，所述反应容器包括弹性部分，所述弹性部分能够发生形变以压迫容置于所述容纳腔中的第一反应试剂或第二反应试剂从所述通孔中流出。
  9. 如权利要求1所述的生化反应装置，其特征在于，所述反应容器包括第一反应部和第二反应部，所述第一反应部开设有第一腔体，所述第二反应部开设有第二腔体，所述第二反应部可拆卸地装设于所述第一反应部上使所述第二腔体和所述第一腔体共同形成一密封腔。
  10. 一种基于CRISPR技术的检测系统，包括：
    如权利要求1-9中任一项所述的生化反应装置；
    第一反应试剂，收容于所述第一反应腔中，用于使待检测样品发生扩增富集反应；以及
    第二反应试剂，收容于所述第二反应腔中，用于使扩增富集产物进行CRISPR检测反应。
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