Reaction device for CRISPR (clustered regularly interspaced short palindromic repeats) test paper detection or Ago protein test paper detection

Abstract

The utility model discloses a reaction device for CRISPR (clustered regularly interspaced short palindromic repeats) test paper detection or Ago protein test paper detection. The reaction device comprises a reaction container, the reaction container comprises a body, the body comprises an opening and an inner cavity, and the opening is formed in one end of the body and connected with the inner cavity; the reaction container further comprises a first partition plate, the inner cavity is divided into a first cavity body and a second cavity body by the first partition plate, and the opening is communicated with the first cavity body; and the overflow hole is formed in the first partition plate and is communicated with the first cavity and the second cavity. The utility model solves the technical problems that the existing CRI SPR test paper detection and Ago protein test paper detection need to be carried out after nucleic acid amplification, so that pollution (such as aerosol pollution) is caused, and false positive results are caused.

Description

TECHNICAL FIELD

[0001] The utility model relates to a reaction device, especially to a reaction device for detection, more especially to a reaction device for CRISPR test paper detection or Ago protein test paper detection. BACKGROUND

[0002] CRISPR detection includes CRISPR nucleic acid detection and CRISPR non-nucleic acid detection. CRISPR nucleic acid detection is the abbreviation of CRISPR / Cas system nucleic acid detection. CRISPR nucleic acid detection is a nucleic acid detection based on the system developed by CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and Cas protein (the abbreviation of CRISPR associated protein). CRISPR nucleic acid detection includes nucleic acid detection using the cis cleavage activity of Cas protein and nucleic acid detection using the trans cleavage activity of Cas protein. The components required for CRISPR nucleic acid detection using the trans cleavage activity of Cas protein include Cas protein, guide RNA, and single-stranded nucleic acid and / or nucleic acid analogue reporter molecule (also known as nucleic acid probe). The core of this nucleic acid detection is still the principle of base pairing, that is, the guide sequence of the guide RNA is paired with the target nucleic acid, and the Cas protein acts on the binding of the direct repeat sequence (DR) of the guide RNA, and after the guide sequence of the guide RNA is paired with the target nucleic acid, it can be activated to cut activity. The activation of the cutting activity is used by people to report signals to obtain qualitative or quantitative information of target nucleic acid detection. Therefore, Cas protein can also be regarded as part of the signal reporting.

[0003] "one-step" refers to the realization of nucleic acid amplification and CRISPR nucleic acid detection in one step. The concept similar to "one-step" is "one-pot" (also known as "one-pot method"). "One-pot" refers to the realization of nucleic acid amplification and CRISPR nucleic acid detection in one container, which may be a two-step method or a one-step method.

[0004] At present, when carrying out two-step CRISPR nucleic acid detection and other two-step reactions, the reagents required for the second step reaction need to be added after the first step reaction is completed.

[0005] The Ago protein detection includes Ago protein nucleic acid detection and Ago protein non-nucleic acid detection. The Ago protein nucleic acid detection refers to nucleic acid detection by using Argonaute protein. For example, a nucleic acid detection method based on prokaryotic Argonaute protein and application thereof are disclosed in Chinese patent CN108796036A, a nucleic acid detection method based on normal-temperature prokaryotic Argonaute protein and application thereof are disclosed in CN114277109A, a visual detection system, reagent or kit for detecting target nucleic acid molecules and a detection method are disclosed in CN114085892A, and a nucleic acid detection method based on medium-temperature Argonaute protein and isothermal amplification is disclosed in CN116064736A. In addition, short pAgo and its related nuclease effect protein can form a heterodimer complex (TmuRE-Ago complex). Unlike long pAgo which specifically cleaves target DNA, the complex is activated after RNA-guided DNA target recognition and exhibits high non-specific DNA cleavage activity (see https: / / doi.org / 10.1093 / nar / gkad1145). This non-specific DNA cleavage activity can also be used for detection.

[0006] Currently, both CRISPR detection and Ago protein detection have test paper detection. However, the current CRISPR detection and Ago protein detection test paper detection, especially the detection of nucleic acid targets, needs to be opened after nucleic acid amplification, which will cause pollution (such as aerosol pollution) and thus the problem of false positives. Utility model content

[0007] The first aspect of the utility model aims to provide a reaction device for CRISPR test paper detection or Ago protein test paper detection, so as to solve the technical problem that the current CRISPR test paper detection and Ago protein test paper detection need to be opened after nucleic acid amplification, which will cause pollution (such as aerosol pollution) and thus the problem of false positives.

[0008] The utility model solves the above technical problems and achieves the technical effects of the utility model by the following technical solutions.

[0009] A reaction device for CRISPR test paper detection or Ago protein test paper detection includes a reaction container, which includes:

[0010] A body includes an opening and an inner cavity, the opening is arranged at one end of the body, and the opening is connected with the inner cavity;

[0011] The reaction container further includes:

[0012] a first partition plate, the first partition plate separating the inner cavity into a first cavity and a second cavity, the opening being in communication with the first cavity;

[0013] an overflow hole, the overflow hole being disposed on the first partition plate, the overflow hole being in communication between the first cavity and the second cavity.

[0014] Preferably, the body is a plastic transparent tubular body, a cuboid body or a conical tubular body.

[0015] Further to any of the above technical solutions, the reaction container further comprises a cover body, the cover body being adapted to the opening of the body. Still further, the reaction container further comprises a connecting portion, one side of the cover body being connected to the body through the connecting portion.

[0016] Further to any of the above technical solutions, the reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises a reagent container, the reagent container being separate from the reaction container, the reagent container comprising a container mouth, the container mouth of the reagent container being adapted to the opening of the body. Still further, the reagent container is a soft plastic reagent bottle, and the container mouth is provided with a sharp nozzle.

[0017] Further to any of the above technical solutions, the reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises:

[0018] a CRISPR detection reagent freeze-dried body or an Ago protein detection reagent freeze-dried body, the CRISPR detection reagent freeze-dried body or the Ago protein detection reagent freeze-dried body being located in the first cavity;

[0019] a CRISPR detection test paper or an Ago protein detection test paper, the CRISPR detection test paper or the Ago protein detection test paper being located in the second cavity and being arranged along the length direction of the reaction container. Still further, the CRISPR detection reagent freeze-dried body or the Ago protein detection reagent freeze-dried body is a CRISPR detection reagent freeze-dried microsphere or an Ago protein detection reagent freeze-dried microsphere with a diameter of 1 mm-5 mm.

[0020] Still further, the reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises:

[0021] a second partition located in the first cavity and above the overflow hole, the second partition partially separates the first cavity into an upper chamber and a lower chamber, the upper chamber and the lower chamber are communicated by a communication hole or a communication groove, the upper chamber is communicated with the opening of the body; the overflow hole communicates the lower chamber with the second cavity; the CRISPR detection reagent freeze-dried microspheres or Ago protein detection reagent freeze-dried microspheres are located in the second cavity and the size is greater than the pore size of the communication hole or the width of the communication groove.

[0022] Further, the reaction container further comprises: a sealing film sealing the opening of the body.

[0023] Further based on any of the above technical solutions, the overflow hole is close to the other end of the body.

[0024] The CRISPR test paper detection reaction container or Ago protein test paper detection reaction container has simple structure, is convenient to use, can complete nucleic acid amplification, CRISPR reaction or Ago protein reaction and test paper strip detection in the container, and does not need to be opened in the middle. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is a structural schematic diagram of the CRISPR test paper detection reaction device or Ago protein test paper detection reaction device of the embodiment of the utility model. DETAILED DESCRIPTION

[0026] TERMS

[0027] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0028] The term "CRISPR" refers to Clustered regularly interspaced short palindromic repeats, which is from the immune system of microorganisms.

[0029] The term "CRISPR-Cas": a unique genomic element derived from bacteria and archaea, as an adaptive immune defense system to resist invading phages or foreign nucleic acids. The system is composed of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (abbreviated as Cas proteins, Cas).

[0030] The term "Cas protein" refers to a CRISPR-associated protein, which is a relevant protein in the CRISPR system. The "Cas protein" described herein refers to a CRISPR-associated protein (sometimes translated as CRISPR-Cas effector protein, CRISPR / Cas effector protein, CRISPR-Cas effector, CRISPR / Cas effector). The Cas proteins currently detected include type I Cas protein (Cas3), type II Cas protein (Cas9), type III Cas protein (Cas10), type V Cas protein (Cas12) or type VI Cas protein (Cas13). In particular, type V Cas protein (Cas12), type VI Cas protein (Cas13) and part of Cas3 and Cas10 are found to have trans-cleavage activity, which can amplify the detection signal, so the trans-cleavage activity is often used for detection. Taking type V Cas protein as an example, once it binds to the cis-cleavage substrate under the guidance of the guide RNA to form a ternary complex of Cas protein-guide RNA-cis-cleavage substrate, it can induce its trans-cleavage activity, i.e. random cleavage of single-stranded DNA (including base-modified single-stranded DNA), and there are also reports that it will randomly cleave single-stranded nucleic acid analogs. Of course, the cis-cleavage activity of Cas protein or other properties can also be used for detection.

[0031] The Cas protein described in the present specific embodiment is preferably a protein having trans-cleavage activity. In particular, the Cas protein still has activity, especially trans-cleavage activity, at a temperature higher than the temperature of the system in which the isothermal amplification reaction is carried out.

[0032] The term "Cas12a" (formerly "Cpf1") is a crRNA-dependent endonuclease, which is a V-A type enzyme in the classification of CRISPR system.

[0033] The term "Cas12b" (formerly "C2c1") is an sgRNA-dependent endonuclease, which is a V-B type enzyme in the classification of CRISPR system.

[0034] The term "PAM" refers to a protospacer-adjacent motif, which is a short DNA sequence directly adjacent to the DNA sequence targeted by the CRISPR effector protein, and is necessary for Cas12a or Cas12b to cleave double-stranded DNA, for example, the PAM of Cas12a is TTTV, and the PAM of AacCas12b is TTN sequence.

[0035] The term "target DNA or RNA molecule" is the DNA or RNA to be detected or its specific part when the nucleic acid molecule to be detected is a nucleic acid molecule; when the non-nucleic acid molecule to be detected is a nucleic acid sequence designed in advance.

[0036] The term "CRISPR nucleic acid detection method" refers to a nucleic acid detection method using a Cas protein, including a nucleic acid detection method using Cas protein cis-cleavage activity, trans-cleavage activity or other functions.

[0037] The term "(one-step CRISPR nucleic acid detection method (using Cas protein trans-cleavage activity))" (or simply CRISPR one-step nucleic acid detection, CRISPR one-step, one-step detection, one-step) is a rapid and convenient detection technology developed on the basis of the CRISPR nucleic acid detection system, which can simultaneously realize the amplification and detection of target nucleic acid in one reaction tube. This technology combines CRISPR-Cas system and isothermal amplification (or constant temperature amplification) technology, and does not need to open the cover operation of the amplified nucleic acid product, which can specifically detect the target nucleic acid in a short time. The CRISPR one-step detection technology is a rapid, accurate, high sensitivity and high specificity detection technology, which not only has simple operation, but also can improve the detection specificity of the current isothermal amplification technology. Compared with the traditional PCR technology, the CRISPR one-step detection does not need complex temperature control and multi-step operation, and has higher real-time and portability. The Chinese invention patent with application publication number CN 110551800 A and application publication date 2019.12.10 first discloses one-step method (see the patent application

[0238] ,

[0239] and other paragraphs).

[0038] The term "system" should be understood broadly, which can be a composition, a product combination, a reagent, a kit, or an instrument containing the aforementioned composition, product combination, reagent, kit, or a mixture (system) formed when the composition, product combination, reagent, kit is used for detection, and an instrument containing the aforementioned mixture, etc.

[0039] The term "temperature" refers to the temperature of the system (the mixture formed when used for detection).

[0040] The term "guide RNA" is a mature crRNA fused (or not fused) with tracrRNA as a guide RNA, or a mature crRNA fused (or not fused) with scoutRNA as a guide RNA, or a crRNA alone as a guide RNA.

[0041] Generally, a guide RNA (gRNA) can comprise, consist essentially of, or consist of direct repeat sequences (also referred to as DR sequences) and a guide sequence. The gRNA can include crRNA and tracrRNA, crRNA and scoutRNA, or only crRNA in different Type V CRISPR systems depending on the Cas protein it relies on. The crRNA and tracrRNA can be artificially engineered to form a single guide RNA (sgRNA). In some cases, the guide sequence is a polynucleotide sequence that is of sufficient complementarity to hybridize to and direct specific binding of a CRISPR / Cas protein-guide RNA complex to a cis-cleavage substrate nucleic acid, typically having a sequence length of 15-28 nt in Type V CRISPR systems. The direct repeat sequences can fold to form a specific structure (e.g., stem-loop structure) for recognition by the Cas protein to form a complex. The guide sequence does not need to be 100% complementary to the cis-cleavage substrate nucleic acid. The guide sequence is not complementary to the nucleic acid in the trans-cleavage reporter molecule.

[0042] In some embodiments, the degree of complementarity (match) between the guide sequence and its corresponding cis-cleavage substrate nucleic acid, when optimally aligned, is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%. Determining optimal alignment is within the capabilities of a person of ordinary skill in the art. For example, there are publicly available and commercially available alignment algorithms and programs, such as, but not limited to, ClustalW, Smith-Waterman in matlab, Bowtie, Geneious, Biopython, and SeqMan.

[0043] The terms “polynucleotide,” “nucleotide sequence,” “nucleic acid sequence,” “nucleic acid molecule,” and “nucleic acid” can be used interchangeably and encompass DNA, RNA, or hybrids thereof, which can be double-stranded or single-stranded, unless indicated otherwise.

[0044] The terms "homology" or "identity" are used in reference to the matching of sequences between two polypeptides or between two nucleic acids. When a position in each of two sequences being compared is occupied by the same base or amino acid monomer subunit (e.g., a position in each of two DNA molecules occupied by adenine, or a position in each of two polypeptides occupied by lysine), then the molecules are identical at that position. Generally, comparisons are made using complete sequences aligned for maximum identity. Such alignments can be determined by computerized running of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group) using, for example, the default parameters of the programs. BLAST algorithms available from the National Center for Biotechnology Information (NCBI www.ncbi.nlm.nih.gov / ) can also be used, using the default parameters.

[0045] The term "nucleic acid analogues" is a class of derivatives of RNA and DNA, which is mainly composed of phosphate, pentose and base, while the nucleic acid analogue replaces at least one of them with some other substance. The main nucleic acid analogues are peptide nucleic acid (PNA), morpholino (MNA), bridged nucleic acid (BNA), locked nucleic acid (LNA), glycol nucleic acid (GNA) and threose nucleic acid (TNA). Some of these nucleic acid analogues can even carry out biological processes such as replication, translation, etc. in vitro (Brudno, Yevgeny; Birnbaum, Michael E; Kleiner, Ralph E; Liu, David R. "An in vitro translation, selection and amplification system for peptide nucleic acids". Nature Chemical Biology. 6 (2): 148-155. doi:10.1038 / nchembio.280. PMC 2808706. PMID 20081830).

[0046] The term "test sample" refers to a sample obtained from a biological sample that has been subjected to extraction of nucleic acids, which sample can also have been subjected to amplification, transcription, reverse transcription of nucleic acids. The biological sample is any solid or fluid sample obtained, excreted or secreted from any organism, including but not limited to unicellular organisms, such as bacteria, yeast, protozoa, and amoebae, and multicellular organisms (e.g., plants or animals, including samples from a healthy or apparently healthy human subject or a human patient affected by a condition or disease to be diagnosed or investigated, e.g., an infection by a pathogenic microorganism, such as a pathogenic bacterium or virus). For example, the biological sample can be a biological fluid obtained from, e.g., blood, plasma, serum, urine, feces, sputum, mucus, lymph, synovial fluid, bile, ascites, pleural effusion, seroma, saliva, cerebrospinal fluid, aqueous or vitreous fluid, or any bodily secretion, exudate, exudate (e.g., fluid obtained from an abscess or any other site of infection or inflammation), or fluid obtained from a joint (e.g., a normal joint or a joint affected by a disease, such as rheumatoid arthritis, osteoarthritis, gout, or septic arthritis), or a swab of a skin or mucosal surface. The sample can also be a sample obtained from any organ or tissue (including a biopsy or autopsy specimen, such as a tumor biopsy) or can comprise cells (primary cells or cultured cells) or a medium conditioned by any cell, tissue, or organ. Exemplary samples include, but are not limited to, cells, cell lysates, blood smears, cytocentrifuged preparations, cytological smears, body fluids (e.g., blood, plasma, serum, saliva, sputum, urine, bronchoalveolar lavage, semen, etc.), tissue biopsies (e.g., tumor biopsies), fine needle aspirates, and / or tissue sections (e.g., cryostat tissue sections and / or paraffin-embedded tissue sections).

[0047] In other embodiments, the biological sample can be a plant cell, callus, tissue, or organ (e.g., roots, stems, leaves, flowers, seeds, fruits), etc.

[0048] The "sample to be tested" can contain a nucleic acid molecule to be tested. In the present application, the nucleic acid molecule to be tested includes a DNA molecule, and also includes an RNA molecule or a DNA molecule formed by reverse transcription of an RNA molecule, or further, the nucleic acid molecule to be tested can be amplified by a technique known in the art, and the amplification technique is an isothermal amplification technique, and the isothermal amplification can be LAMP (loop-mediated isothermal amplification), RPA (recombinase polymerase amplification), RAA (recombinase-mediated amplification), ERA (enzyme recombination isothermal amplification technique), MIRA (multi-enzyme isothermal rapid amplification technique), bDNA (branch DNA amplification), NASBA (nucleic acid sequence-based amplification), SDA (strand displacement amplification), TMA (transcription-mediated amplification), RCA (rolling circle amplification), HDA (helicase-dependent amplification), SPIA (single primer isothermal amplification), NEAR (nicking enzyme amplification reaction), SMAP (smart amplification process), SMAP2 (second edition of smart amplification process), CPA (crossover primer amplification), MDA (multiple displacement amplification), RAM (Ramification), cHDA (helicase-dependent circular amplification), SMART (signal-mediated amplification of RNA technique), 3SR (self-sustained sequence replication system), GEAR (genomic exponential amplification reaction), IMDA (isothermal multiple displacement amplification), ERA (enzyme recombination isothermal amplification), TAS (transcription-dependent amplification system), RIDA (rapid isothermal detection amplification technique), NEMA (nicking endonuclease isothermal amplification of nucleic acids), EXPAR (exponential isothermal amplification), ICAN (isothermal chimeric primer-initiated amplification of nucleic acids), SEA (strand exchange amplification), SHARP (SSB-helicase-mediated rapid PCR), IMSA (isothermal multiple self-priming amplification), WGA (whole genome amplification), PSR (polymerase spiral reaction), or a combination thereof.

[0049] Further, the detection method of the present application further comprises a step of amplifying the nucleic acid molecule to be tested; and the detection system further comprises components for amplifying the nucleic acid molecule to be tested. The components for amplification include one or more of the following: a DNA polymerase, a reverse transcriptase, a strand displacement enzyme, a nicking endonuclease, a helicase, a recombinase, a single-strand binding protein, a recombination modulating protein, a T7 RNA polymerase, an RNase H, dNTPs for amplification reaction and / or reverse transcription reaction, NTPs for transcription reaction, a buffer, and the like.

[0050] The term "Ago protein" is an Argonaute protein.

[0051] The term "Ago protein nucleic acid detection" refers to nucleic acid detection using Argonaute protein, such as "Nucleic acid detection method based on prokaryotic Argonaute protein and its application" disclosed in Chinese invention patent CN108796036A, "Nucleic acid detection method based on normal temperature prokaryotic Argonaute protein and its application" disclosed in CN114277109A, "Visual detection system, reagent or kit for detecting target nucleic acid molecules and detection method" disclosed in CN114085892A, and "Nucleic acid detection method based on medium temperature Argonaute protein and isothermal amplification" disclosed in CN116064736A. In addition, short pAgo and its related nuclease effect protein can form a heterodimer complex (TmuRE-Ago complex); unlike long pAgo which specifically cuts target DNA, this complex is activated after RNA-guided DNA target recognition, showing high non-specific DNA cleavage activity (see https: / / doi.org / 10.1093 / nar / gkad1145). This non-specific DNA cleavage activity can also be used for detection.

[0052] Referring to Figure 1 A reaction device for CRISPR test paper detection or Ago protein test paper detection includes a reaction container 1 and a reagent container 2; it can also include a preloaded CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body 3, a CRISPR detection test paper or an Ago protein detection test paper 4. The reagent container can be preloaded with liquid reagent, or the sample to be tested can be loaded at the time of use.

[0053] The reaction container 1 includes a body 11, a first partition plate 12, a first cavity 101, a second cavity 102, and an overflow hole 13; it can also include a cover 14 and a connecting part 15. The body 11 includes an opening 10 and an inner cavity, the opening 10 is arranged at one end of the body and is connected to the inner cavity. The first partition plate 12 divides the inner cavity into the first cavity 101 and the second cavity 102, and the opening is in communication with the first cavity 101; the overflow hole 13 is arranged on the first partition plate 12, and is in communication with the first cavity 101 and the second cavity 102, and the overflow hole 13 is close to the other end of the body 11. Figure 1 The body 11 is a plastic transparent cuboid body, which can also be a plastic transparent tubular body or a plastic transparent conical tubular body. The plastic transparent material is conducive to observation.

[0054] The cover 14 is matched with the opening 10 of the body, that is, it can tightly cover the opening. One side of the cover 14 is connected to the body 11 through the connecting part 15.

[0055] The reagent container 2 is free from the reaction container 1, and the reagent container 2 comprises a container mouth which is matched with the opening 10 of the body 11, that is, the reagent container 2 can be arranged in the opening 10, which can be threadedly connected, and the connection is tight so that nucleic acid cannot overflow. The reagent container 2 is preferably a soft plastic reagent bottle, and the container mouth is provided with a sharp mouth 21, so that after the reagent container 2 is inserted into the opening 10 of the body 11 of the reaction container 1, the liquid reagent and the sample to be tested contained in the reagent container cannot fall into the reaction container 1, and the liquid reagent can be added into the reaction container 1 by extruding the soft plastic reagent bottle.

[0056] The CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body 3 is located in the first cavity 101, and can be preloaded in the first cavity 101 in the form of in-situ freeze-drying; the CRISPR detection test paper or Ago protein detection test paper 4 is located in the second cavity 102 and is arranged along the length direction of the reaction container 1, and one side of the body 11 can be arranged as a structure which can be opened and closed, so as to facilitate preloading of the CRISPR detection test paper or Ago protein detection test paper 4, which belongs to the prior art and will not be described here.

[0057] In addition to in-situ freeze-drying, the CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body 3 can also be a CRISPR detection reagent freeze-dried microsphere or Ago protein detection reagent freeze-dried microsphere with a diameter of 1 mm-5 mm. In this technical solution, in order to prevent the CRISPR detection reagent freeze-dried microsphere or Ago protein detection reagent freeze-dried microsphere from running away after preloading, the reaction device for CRISPR test paper detection or Ago protein test paper detection can further comprise:

[0058] The second partition plate 16 is located in the first cavity 101 and above the overflow hole 13, and the second partition plate 16 partially divides the first cavity 101 into an upper chamber 1011 and a lower chamber 1012, the upper chamber 1011 and the lower chamber 1012 are communicated through a communication hole or a communication groove 17, and the upper chamber 1011 is communicated with the opening 10 of the body 11; the overflow hole 13 communicates the lower chamber 1012 with the second cavity 102; the CRISPR detection reagent freeze-dried microsphere or Ago protein detection reagent freeze-dried microsphere is located in the lower chamber 1012 and has a size greater than the pore diameter of the communication hole or the width of the communication groove 17.

[0059] In the case of preloading the CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body, the reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises a sealing film (not shown in the figure) which seals the opening 10 of the body 11. Of course, the CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body can also not be preloaded, and can be loaded from the opening when in use. In this technical solution, the second partition plate 16 does not need to be provided.

[0060] The utility model is explained below from the use method. With the reagent container 2 of preloaded CRISPR detection reagent freeze-dried body or Ago protein detection reagent freeze-dried body 3, reagent soft plastic bottle is taken as an example, when using, the liquid reagent (such as diluent etc.) of reagent container 2 is loaded, and the sample to be measured, the sharp mouth 21 is sleeved on the container mouth of reagent container 2, the sealing film on the opening 10 of the body 11 of reaction container 1 is punctured, reagent container 2 is inserted into opening 10 (the container mouth of reagent container 2 and the opening 10 of the body 11 are provided with screw threads, then the reagent container 2 is squeezed, and the first cavity 101 is dripped into part of liquid reagent, the sample to be measured, after the time required for reaction, the reagent container 2 is continuously squeezed, and part of liquid reagent, the sample to be measured is continuously dripped, until the reaction product in reaction container 1 can flow into the second cavity 102 from the overflow hole 13, after the time required for test paper detection, the test paper is observed, and whether the target exists in the sample to be measured is judged.

[0061] Various changes can be made according to the disclosure of the present specification by those skilled in the art after reading the present specification, and the changes all belong to the utility model. The description of the present specification does not limit the protection scope of the utility model, and the protection scope of the utility model is subject to the claims and equivalent embodiments.

Claims

1. A reaction device for CRISPR test paper detection or Ago protein test paper detection, comprising a reaction container, the reaction container comprising: a body, the body comprising an opening and an inner cavity, the opening being arranged at one end of the body and connecting the inner cavity; characterized in that the reaction container further comprises: a first partition plate, the first partition plate separating the inner cavity into a first cavity and a second cavity, the opening being in communication with the first cavity; an overflow hole, the overflow hole being arranged on the first partition plate and communicating the first cavity and the second cavity.

2. The reaction device for CRISPR test paper detection or Ago protein test paper detection according to claim 1, characterized in that, The body is a plastic transparent tubular body, a cuboid body or a conical tubular body.

3. The reaction device for CRISPR test paper or Ago protein test paper according to claim 1 or 2, characterized in that, The reaction container further comprises a cover body, the cover body being matched with the opening of the body.

4. The reaction device for CRISPR test paper or Ago protein test paper according to claim 1 or 2, characterized in that, The reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises a reagent container, the reagent container being separated from the reaction container, the reagent container comprising a container mouth, the container mouth of the reagent container being matched with the opening of the body.

5. The reaction device for CRISPR test paper or Ago protein test paper according to claim 4, characterized by, The reagent container is a soft plastic reagent bottle, and the container mouth is provided with a sharp nozzle.

6. The reaction device for CRISPR test paper or Ago protein test paper according to claim 1 or 2, characterized in that, The reaction device for CRISPR test paper detection or Ago protein test paper detection further comprises: a CRISPR detection reagent freeze-dried body or an Ago protein detection reagent freeze-dried body, the CRISPR detection reagent freeze-dried body or the Ago protein detection reagent freeze-dried body being located in the first cavity; a CRISPR detection test paper or an Ago protein detection test paper, the CRISPR detection test paper or the Ago protein detection test paper being located in the second cavity and arranged along the length direction of the reaction container.

7. The reaction device for CRISPR test paper or Ago protein test paper according to claim 6, wherein The CRISPR detection reagent freeze-dried body or the Ago protein detection reagent freeze-dried body is a CRISPR detection reagent freeze-dried microsphere or an Ago protein detection reagent freeze-dried microsphere with a diameter of 1 mm-5 mm.

8. The reaction device for CRISPR test paper or Ago protein test paper according to claim 7, wherein Further comprising: a second partition plate, the second partition plate being located in the first cavity and above the overflow hole, the second partition plate partially separating the first cavity into an upper chamber and a lower chamber, the upper chamber and the lower chamber being in communication through a communication hole or a communication groove, the upper chamber being in communication with the opening of the body; the overflow hole communicating the lower chamber and the second cavity; the CRISPR detection reagent freeze-dried microsphere or the Ago protein detection reagent freeze-dried microsphere being located in the second cavity and having a size greater than the pore diameter of the communication hole or the width of the communication groove.

9. The reaction device for CRISPR test paper or Ago protein test paper according to claim 6, wherein The reaction container further comprises a sealing film, the sealing film sealing the opening of the body.

10. The reaction device for CRISPR test paper or Ago protein test paper according to claim 1 or 2, characterized in that, The overflow hole is close to the other end of the body.

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