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

Abstract

The utility model relates to a detection device, in particular to a detection device for CRI SPR test paper detection or Ago protein test paper detection. A detection device for CRI SPR test paper detection or Ago protein test paper detection is technically characterized by comprising a containing cavity, and the containing cavity comprises a first cavity, a second cavity and a third cavity, the second cavity is used for accommodating a CRI SPR nucleic acid detection reagent or an Ago protein detection reagent; the third cavity is used for accommodating test paper for CRI SPR detection or test paper for Ago protein detection; openings of the first cavity, the second cavity and the third cavity face the opening of the containing cavity, and the openings of the first cavity, the second cavity and the third cavity are communicated at the opening of the containing 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] This utility model relates to a detection device, and more particularly to a detection device for CRISPR test strip detection or Ago protein test strip detection. Background Technology

[0002] CRISPR assays include CRISPR nucleic acid assays and CRISPR non-nucleic acid assays. CRISPR nucleic acid assays are short for CRISPR / Cas system nucleic acid assays. CRISPR nucleic acid assays are nucleic acid assays developed based on the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and Cas protein (CRISPR associated protein) system. CRISPR nucleic acid assays include those utilizing the cis-cleavage activity of the Cas protein and those utilizing the trans-cleavage activity of the Cas protein. The components required for CRISPR nucleic acid assays utilizing the trans-cleavage activity of the Cas protein include the Cas protein, guide RNA, and single-stranded nucleic acid and / or nucleic acid analog reporter molecules (also referred to as nucleic acid probes in some literature). The core of this nucleic acid assay remains the base pairing principle: the guide sequence of the guide RNA pairs with the target nucleic acid. The role of the Cas protein is to bind to the direct repeat (DR) sequence of the guide RNA, and after the guide sequence of the guide RNA pairs with the target nucleic acid, its cleavage activity is activated. Activation of cleavage activity has been utilized for signal reporting to obtain qualitative or quantitative information for target nucleic acid detection. Therefore, Cas proteins can also be considered as part of signal reporting.

[0003] "One-step" refers to the process of performing nucleic acid amplification and CRISPR nucleic acid detection in a single step. A similar concept is "one-pot" (also known as "one-tube method"). One-pot method involves performing nucleic acid amplification and CRISPR nucleic acid detection in a single container; this can be either a two-step or one-step process.

[0004] Currently, in experiments involving two-step reactions such as the CRISPR nucleic acid detection two-step method, it is necessary to open the lid and add the reagents required for the second step reaction after completing the first step reaction.

[0005] Ago protein detection includes Ago protein nucleic acid detection and Ago protein non-nucleic acid detection. "Ago protein nucleic acid detection" refers to nucleic acid detection using Argonaute protein, such as the "Nucleic Acid Detection Method and Its Application Based on Prokaryotic Argonaute Protein" disclosed in Chinese invention patent CN108796036A, the "Nucleic Acid Detection Method and Its Application Based on Room Temperature Prokaryotic Argonaute Protein" disclosed in CN114277109A, the "Visual Detection System, Reagent or Kit and Detection Method for Detecting Target Nucleic Acid Molecules" disclosed in CN114085892A, and the "Nucleic Acid Detection Method Based on Mesothermal Argonaute Protein and Isothermal Amplification" disclosed in CN116064736A. Furthermore, short pAgo and its associated nuclease effector proteins can form a heterodimeric complex (TmuRE-Ago complex); unlike long pAgo which specifically cleaves target DNA, this complex is activated upon RNA-guided DNA target recognition, exhibiting highly efficient 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 and Ago protein assays have developed test strips. However, current CRISPR and Ago protein assays, especially for nucleic acid targets, require opening the test strip after nucleic acid amplification, which can lead to contamination (such as aerosol contamination) and thus false positives. Utility Model Content

[0007] The first objective of this invention is to provide a detection device for CRISPR test strips or Ago protein test strips, in order to solve the technical problem that current CRISPR test strip and Ago protein test strip detection require opening the cap after nucleic acid amplification, which causes contamination (such as aerosol contamination) and thus leads to false positives.

[0008] The present invention solves the above-mentioned technical problems through the following technical solutions, thereby achieving the technical effects of the present invention.

[0009] A detection device for CRISPR test strip detection or Ago protein test strip detection includes: a accommodating cavity, the accommodating cavity including an opening and an inner cavity, one end of the accommodating cavity being provided with the opening and the opening being connected to the inner cavity;

[0010] A cover body that is adapted to the opening;

[0011] The accommodating cavity includes:

[0012] A first cavity for containing the diluent, the first cavity being separated from the inner cavity;

[0013] A second cavity is provided for accommodating CRISPR nucleic acid detection reagents or Ago protein detection reagents, the second cavity being separated from the inner cavity;

[0014] A third cavity is provided for accommodating CRISPR detection strips or Ago protein detection strips, the third cavity being separated from the inner cavity;

[0015] The openings of the first cavity, the second cavity, and the third cavity face the opening of the receiving cavity, and the openings of the first cavity, the second cavity, and the third cavity communicate with each other at the opening of the receiving cavity.

[0016] Preferably, the accommodating cavity is a tubular accommodating cavity, a cuboid accommodating cavity, or a conical tubular accommodating cavity.

[0017] Preferably, the top of the cover is flat, the lower part of the cover can be threaded or snapped to one end of the accommodating cavity, and one side of the cover is connected to the accommodating cavity through a connecting part.

[0018] Preferably, the ratio of the cross-sectional area of ​​the cover to the cross-sectional area of ​​the accommodating cavity is 1.1:1-10:1.

[0019] Furthermore, the detection device for CRISPR test strip detection or Ago protein test strip detection also includes:

[0020] First partition;

[0021] Second partition;

[0022] The first partition and the second partition divide the inner cavity of the accommodating cavity into the first cavity, the second cavity, and the third cavity; the third cavity is located to the left of the second cavity, and the first cavity is located to the right of the second cavity.

[0023] Furthermore, the detection device for CRISPR test strip detection or Ago protein test strip detection also includes: a sealing film, which seals the opening of the accommodating cavity.

[0024] Preferably, the accommodating cavity is a tubular accommodating cavity made of transparent plastic.

[0025] Furthermore, the detection device for CRISPR test strip detection or Ago protein test strip detection also includes:

[0026] CRISPR nucleic acid detection reagent or Ago protein detection reagent, wherein the CRISPR nucleic acid detection reagent or Ago protein detection reagent is contained in the second cavity;

[0027] CRISPR detection strips or Ago protein detection strips are housed within the third cavity, and the CRISPR detection strips or Ago protein detection strips are arranged along the length of the third cavity.

[0028] Preferably, the CRISPR nucleic acid detection reagent or Ago protein detection reagent is a lyophilized microsphere with a diameter of 1 mm-5 mm or is disposed in the second cavity in the form of in-situ lyophilization. The CRISPR nucleic acid detection reagent or Ago protein detection reagent is a one-step CRISPR nucleic acid detection reagent lyophilized body or a one-step Ago protein nucleic acid detection reagent lyophilized body.

[0029] The detection device for CRISPR test strip detection or Ago protein test strip detection of this utility model has a simple structure and is easy to use. It can complete nucleic acid amplification, CRISPR reaction or Ago protein reaction and test strip detection in the container. There is no need to open the lid in the middle. The whole process can be completed by simply inverting the detection device. Attached Figure Description

[0030] Figure 1 This is a schematic diagram of the detection device for CRISPR test strip detection or Ago protein test strip detection according to a specific embodiment of this utility model. Detailed Implementation

[0031] the term

[0032] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0033] The term "CRISPR" refers to clustered regularly interspaced short palindromic repeats, which originate from the immune system of microorganisms.

[0034] The term "CRISPR-Cas" refers to a unique genomic element derived from bacteria and archaea, serving as an adaptive immune defense system against invading bacteriophages or foreign nucleic acids. This system consists of clusters of regularly spaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas proteins, or Cas for short).

[0035] The term "Cas protein" refers to CRISPR-associated proteins, which are related proteins in the CRISPR system. In this article, "Cas protein" refers to CRISPR-related proteins (sometimes translated as CRISPR-Cas effector proteins, CRISPR / Cas effector proteins, CRISPR-Cas effectors, or CRISPR / Cas effectors). Currently used Cas proteins for detection include type I Cas protein (Cas3), type II Cas protein (Cas9), type III Cas protein (Cas10), type V Cas protein (Cas12), and type VI Cas protein (Cas13). In particular, type V Cas protein (Cas12), type VI Cas protein (Cas13), and some Cas3 and Cas10 proteins have been found to have trans-cleavage activity, which can amplify the detection signal; therefore, their trans-cleavage activity is commonly used for detection. Taking type V Cas protein as an example, once it binds to a cis-cleavage substrate under the guidance of guide RNA to form a ternary complex of Cas protein-guide RNA-cis-cleavage substrate, its trans-cleavage activity can be induced, i.e., randomly cleaving single-stranded DNA (including single-stranded DNA with base modifications). There are also reports of it randomly cleaving single-stranded nucleic acid analogs. Of course, the cis-cleavage activity or other properties of Cas protein can also be used for detection.

[0036] The Cas protein described in this specific embodiment is preferably a protein with trans-cleavage activity. In particular, it is a Cas protein that retains activity, especially trans-cleavage activity, at temperatures higher than the system temperature at which the isothermal amplification reaction is performed.

[0037] The term "Cas12a" (formerly "Cpf1") is a crRNA-dependent endonuclease, which is a type VA enzyme in the CRISPR system classification.

[0038] The term "Cas12b" (formerly "C2c1") is an sgRNA-dependent endonuclease, which is a type VB enzyme in the CRISPR system.

[0039] The term "PAM" refers to the protospacer-adjacent motif, which is a short DNA sequence directly adjacent to the DNA sequence targeted by CRISPR effector proteins. It is essential for Cas12a or Cas12b to cleave double-stranded DNA. For example, the PAM of Cas12a is TTTV, and the PAM of AacCas12b is the TTN sequence.

[0040] The term "target DNA or RNA molecule" refers to the DNA or RNA to be tested or a specific portion thereof when the molecule to be tested is a nucleic acid molecule; when the molecule to be tested is a non-nucleic acid molecule, the target DNA or RNA molecule is a pre-designed nucleic acid sequence.

[0041] The term "CRISPR nucleic acid detection method" refers to nucleic acid detection methods that utilize Cas proteins, including nucleic acid detection methods that utilize the cis-cleavage activity, trans-cleavage activity, or other functions of Cas proteins.

[0042] The term "one-step CRISPR nucleic acid detection (method) utilizing the trans-cleavage activity of Cas proteins" (or simply CRISPR one-step nucleic acid detection, CRISPR one-step, one-step detection, one-step method) is a rapid and convenient detection technology developed based on the CRISPR nucleic acid detection system. It allows for the simultaneous amplification and detection of target nucleic acids in a single reaction tube. This technology combines the CRISPR-Cas system with isothermal amplification (or isothermal amplification) technology, eliminating the need to open the amplified nucleic acid product and enabling specific detection of target nucleic acids within a short time. CRISPR one-step detection technology is a rapid, accurate, highly sensitive, and highly specific detection technique. It is not only easy to operate but also improves upon the detection specificity of current isothermal amplification techniques. Compared to traditional PCR technology, CRISPR one-step detection does not require complex temperature control and multi-step operations, offering greater real-time performance and portability. Chinese invention patent application publication number CN110551800A, with an application publication date of December 10, 2019, first disclosed a one-step method (see paragraphs

[0238] ,

[0239] , etc. of the patent application).

[0043] The term "system" should be interpreted broadly, and can refer to compositions, product combinations, reagents, kits, instruments and equipment containing the aforementioned compositions, product combinations, reagents, kits, mixtures (systems) formed when the compositions, product combinations, reagents, kits are used for detection, as well as instruments and equipment containing the aforementioned mixtures, etc.

[0044] The term "temperature" refers to the temperature of the system (the mixture formed during testing).

[0045] The term "guide RNA" refers to a mature crRNA fused with tracrRNA (or not fused) as a guide RNA, or a mature crRNA fused with scoutRNA (or not fused) as a guide RNA, or crRNA alone as a guide RNA.

[0046] Generally, guide RNA (gRNA) can contain direct repeat sequences (DR sequences) and a guide sequence, or consist primarily of or composed of direct repeat sequences and a guide sequence (also called a spacer sequence in the context of endogenous CRISPR systems). In different type V CRISPR systems, depending on the Cas protein it relies on, gRNA can include crRNA and tracrRNA, crRNA and scoutRNA, or only crRNA. crRNA and tracrRNA can be artificially fused to form single guide RNA (sgRNA). In some cases, the guide sequence is a polynucleotide sequence that is sufficiently complementary to the cis-cleaved substrate nucleic acid to hybridize with it and guide the specific binding of the CRISPR / Cas protein-guide RNA complex to the cis-cleaved substrate nucleic acid. In type V CRISPR systems, it typically has a sequence length of 15-28 nt. The direct repeat sequences can fold into specific structures (such as stem-loop structures) for Cas protein recognition to form a complex. The guide sequence does not need to be 100% complementary to the cis-cleaved substrate nucleic acid. The guide sequence is not complementary to the nucleic acid in the trans-cleaved reporter molecule.

[0047] In some implementations, when optimal alignment is achieved, the complementarity (match) between the guide sequence and its corresponding cis-cleaved substrate nucleic acid 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 skilled in the art. For example, publicly available and commercially available alignment algorithms and programs exist, such as, but not limited to, ClustalW, the Smith-Waterman algorithm in MATLAB, Bowtie, Geneious, Biopython, and SeqMan. The terms “polynucleotide,” “nucleotide sequence,” “nucleic acid sequence,” “nucleic acid molecule,” and “nucleic acid” are used interchangeably and include DNA, RNA, or hybrids thereof, which, unless otherwise specified, may be double-stranded or single-stranded.

[0048] The term "homology" or "identity" is used to refer to the sequence matching between two polypeptides or two nucleic acids. Two compared sequences are considered identical at that position when a position is occupied by the same base or amino acid monomeric 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). Typically, two sequences are compared to produce the greatest possible identity. Such alignments can be determined using, for example, the identity of amino acid sequences, through conventional methods, referring to the teachings of, for example, Smithand Waterman, 1981, Adv. Appl. Math. 2:482, Pearson & Lipman, 1988, Pro. Natl. Acad. Sci. USA 85:244, Thompson et al., 1994, Nucleic Acids Res 22:467380, etc., by computerizing algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group). Alternatively, the BLAST algorithm, available from the National Center for Biotechnology Information (NCBI www.ncbi.nlm.nih.gov / ), can be used with default parameters.

[0049] The term "nucleic acid analogues" refers to a class of RNA and DNA derivatives. Nucleic acids are mainly composed of phosphate, pentose sugar, and bases, while nucleic acid analogues replace at least one of these components with other substances. The main nucleic acid analogues include 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 analogs can even undergo biological processes such as replication and translation in vitro (Brudno, Yevgeny; Birnbaum, Michael E; Kleiner, Ralph E; Liu, David R. "An in vitro translation, selection and amplification system for peptidenucleic acids". Nature Chemical Biology. 6(2): 148–155. doi: 10.1038 / nchembio.280.PMC 2808706.PMID 20081830).

[0050] The term "sample to be tested" refers to a sample obtained by extracting nucleic acids from a biological sample, which may also be obtained through nucleic acid amplification, transcription, or reverse transcription. The biological sample is any solid or fluid sample obtained, excreted, or secreted from any organism, including but not limited to single-celled organisms such as bacteria, yeast, protozoa, and amoebas, and multicellular organisms (e.g., plants or animals, including samples from healthy or seemingly healthy human subjects or human patients affected by a condition or disease to be diagnosed or investigated, such as infections caused by pathogenic microorganisms such as pathogenic bacteria or viruses). For example, a biological sample can be a biological fluid obtained from, for example, 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, biological fluid (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 disease, such as rheumatoid arthritis, osteoarthritis, gout, or septic arthritis), or a swab from the surface of the skin or mucous membrane. The sample can also be a sample obtained from any organ or tissue (including biopsy or autopsy specimens, such as tumor biopsies) or may contain cells (primary cells or cultured cells) or a culture medium conditioning any cell, tissue, or organ. Exemplary samples include, but are not limited to, cells, cell lysates, blood smears, cell centrifugation 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).

[0051] In other embodiments, the biological sample may be plant cells, callus, tissue or organ (such as root, stem, leaf, flower, seed, fruit), etc.

[0052] The "sample to be tested" may contain the nucleic acid molecules to be tested. In this invention, the nucleic acid molecules to be tested include DNA molecules, and also include RNA molecules or DNA molecules formed through reverse transcription of RNA. Furthermore, the nucleic acid molecules to be tested can be amplified using techniques known in the art, specifically isothermal amplification techniques. Isothermal amplification can include LAMP (loop-mediated isothermal amplification), RPA (recombinase polymerase amplification), RAA (recombinase-mediated amplification), ERA (enzyme-catalyzed recombination isothermal amplification), MIRA (multi-enzyme isothermal rapid amplification), bDNA (branched DNA amplification), NASBA (nucleic acid sequence-dependent 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 method), and SMAP2 (version 2). Intelligent amplification methods, CPA (cross-primer amplification), MDA (multiple substitution amplification), RAM (Ramification), cHDA (helicase-dependent circular amplification), SMART (RNA signal-mediated amplification), 3SR (autonomous sequence replication system), GEAR (genomic exponential amplification reaction), IMDA (isothermal multiple substitution amplification), ERA (enzyme-catalyzed recombination isothermal amplification), TAS (transcription-dependent amplification system), RIDA (rapid isothermal detection amplification), NEMA (nicking endonuclease isothermal amplification of nucleic acids), EXPAR (exponential isothermal amplification), ICAN (chimeric primer-induced isothermal amplification of nucleic acids), SEA (strand exchange amplification), SHARP (SSB-helicase-mediated rapid PCR), IMSA (isothermal multiple self-combination amplification), WGA (whole genome amplification), PSR (polymerase helical reaction), or combinations thereof.

[0053] Furthermore, the detection method of the present invention further includes a step of amplifying the nucleic acid molecule to be tested; the detection system further includes components for amplifying the nucleic acid molecule to be tested. The amplification components include one or more of the following: DNA polymerase, reverse transcriptase, strand displacement enzyme, nicking endonuclease, helicase, recombinase, single-strand binding protein, recombinant regulatory protein, T7 RNA polymerase, RNase H, dNTPs for amplification and / or reverse transcription reactions, NTPs for transcription reactions, buffer solutions, etc.

[0054] The term "Ago protein" refers to Argonaute protein.

[0055] The term "Ago protein nucleic acid detection" refers to nucleic acid detection using Argonaute proteins, such as the "Nucleic Acid Detection Method and Its Application Based on Prokaryotic Argonaute Protein" disclosed in Chinese invention patent CN108796036A, the "Nucleic Acid Detection Method and Its Application Based on Room Temperature Prokaryotic Argonaute Protein" disclosed in CN114277109A, the "Visual Detection System, Reagent or Kit and Detection Method for Detecting Target Nucleic Acid Molecules" disclosed in CN114085892A, and the "Nucleic Acid Detection Method Based on Mesothermal Argonaute Protein and Isothermal Amplification" disclosed in CN116064736A. Furthermore, short pAgo and its associated nuclease effector proteins can form a heterodimeric complex (TmuRE-Ago complex); unlike long pAgo which specifically cleaves target DNA, this complex is activated after RNA-guided DNA target recognition, exhibiting highly efficient 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.

[0056] See Figure 1 A detection device for CRISPR nucleic acid detection or Ago protein detection includes a accommodating cavity 1, a cover 2, a first partition 3, and a second partition 4. It may also pre-load CRISPR nucleic acid detection reagents or Ago protein detection reagents 5, and CRISPR detection test strips or Ago protein detection test strips 6. Alternatively, the CRISPR nucleic acid detection reagents or Ago protein detection reagents 5, and CRISPR detection test strips or Ago protein detection test strips 6 may not be pre-loaded and can be loaded during use.

[0057] The receiving cavity 1 is a tubular receiving cavity, a cuboid receiving cavity, or a conical tubular receiving cavity. The receiving cavity 1 includes an opening and an inner cavity. One end of the receiving cavity 1 is provided with an opening that connects to the inner cavity; the cover 2 is adapted to the opening.

[0058] The accommodating cavity 1 includes: a first cavity 11 for containing diluent, a second cavity 12 for containing CRISPR nucleic acid detection reagents or Ago protein detection reagents, and a third cavity 13 for containing CRISPR detection strips or Ago protein detection strips. A first partition 3 and a second partition 4 divide the inner cavity of the accommodating cavity 1 into the first cavity 11, the second cavity 12, and the third cavity 13; the third cavity 13 is located to the left of the second cavity 12, and the first cavity 11 is located to the right of the second cavity 12.

[0059] The openings of the first cavity 11, the second cavity 12, and the third cavity 13 face the opening of the receiving cavity 1, and the openings of the first cavity 11, the second cavity 12, and the third cavity 13 communicate with each other at the opening of the receiving cavity 1. In this way, the reagents in the first cavity 11 and the second cavity 12 can reach the third cavity 13 after the detection device for CRISPR test strip detection or Ago protein test strip detection is placed upside down.

[0060] To facilitate the inverted placement of the detection device for CRISPR test strips or Ago protein test strips, the top of the cover 2 is flat, and the lower part of the cover 2 can be threaded or snapped to one end of the accommodating cavity 1. One side of the cover 2 is connected to the accommodating cavity 1 through a connecting part (not shown in the figure).

[0061] The CRISPR nucleic acid detection reagent or Ago protein detection reagent 5 is contained in the second chamber. The CRISPR nucleic acid detection reagent or Ago protein detection reagent is a lyophilized form for CRISPR detection or a lyophilized form for Ago protein detection.

[0062] CRISPR detection test strip or Ago protein detection test strip 6 is housed in the third cavity and is arranged along the length of the third cavity 13.

[0063] The CRISPR nucleic acid detection reagent or Ago protein detection reagent 5 is a lyophilized microsphere with a diameter of 1 mm-5 mm or is set in the second cavity 12 in the form of in-situ lyophilization. The CRISPR nucleic acid detection reagent or Ago protein detection reagent is preferably a one-step CRISPR nucleic acid detection reagent lyophilized body or a one-step Ago protein nucleic acid detection reagent lyophilized body.

[0064] When the detection device for CRISPR test strip detection or Ago protein test strip detection of this utility model is pre-loaded with a one-step CRISPR nucleic acid detection reagent lyophilized body or a one-step Ago protein nucleic acid detection reagent lyophilized body, a CRISPR detection test strip or an Ago protein detection test strip 6, the detection device also includes a sealing film (not shown in the figure), which seals the opening of the accommodating cavity 1.

[0065] In practical use, remove the sealing film, add the sample to be tested to the second chamber 12, add the diluent (the diluent is the reagent required for the test strip) to the first chamber 11, and tighten the cap 2. Place the detection device for CRISPR test strips or Ago protein test strips in a constant temperature water bath. After the nucleic acid amplification is complete, invert the detection device for CRISPR test strips or Ago protein test strips, wait for a period of time, and observe the test strip to determine whether the target nucleic acid is present.

Claims

1. A detection device for CRISPR test strip detection or Ago protein test strip detection, comprising: A receiving cavity, the receiving cavity including an opening and an inner cavity, wherein one end of the receiving cavity is provided with the opening and the opening is connected to the inner cavity; A cover body that is adapted to the opening; The accommodating cavity is characterized by comprising: A first cavity for containing the diluent, the first cavity being separated from the inner cavity; A second cavity is provided for accommodating CRISPR nucleic acid detection reagents or Ago protein detection reagents, the second cavity being separated from the inner cavity; A third cavity is provided for accommodating CRISPR detection strips or Ago protein detection strips, the third cavity being separated from the inner cavity; The openings of the first cavity, the second cavity, and the third cavity face the opening of the receiving cavity, and the openings of the first cavity, the second cavity, and the third cavity communicate with each other at the opening of the receiving cavity.

2. The detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1, characterized in that, The accommodating cavity is a tubular accommodating cavity, a cuboid accommodating cavity, or a conical tubular accommodating cavity.

3. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The top of the cover is flat, and the lower part of the cover can be threaded or snapped to one end of the accommodating cavity. One side of the cover is connected to the accommodating cavity through a connecting part.

4. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The ratio of the cross-sectional area of ​​the cover to the cross-sectional area of ​​the accommodating cavity is 1.1:1-10:

1.

5. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The detection device for CRISPR test strip detection or Ago protein test strip detection also includes: First partition; Second partition; The first partition and the second partition divide the inner cavity of the accommodating cavity into the first cavity, the second cavity, and the third cavity; the third cavity is located to the left of the second cavity, and the first cavity is located to the right of the second cavity.

6. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The detection device for CRISPR test strip detection or Ago protein test strip detection further includes: a sealing film, which seals the opening of the accommodating cavity.

7. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The accommodating cavity is a tubular accommodating cavity made of transparent plastic.

8. A detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 1 or 2, characterized in that, The detection device for CRISPR test strip detection or Ago protein test strip detection also includes: CRISPR nucleic acid detection reagent or Ago protein detection reagent, wherein the CRISPR nucleic acid detection reagent or Ago protein detection reagent is contained in the second cavity; CRISPR detection strips or Ago protein detection strips are housed within the third cavity, and the CRISPR detection strips or Ago protein detection strips are arranged along the length of the third cavity.

9. The detection device for CRISPR test strip detection or Ago protein test strip detection as described in claim 8, characterized in that, The CRISPR nucleic acid detection reagent or Ago protein detection reagent is a lyophilized microsphere with a diameter of 1 mm-5 mm or is disposed in the second cavity in the form of in-situ lyophilization. The CRISPR nucleic acid detection reagent or Ago protein detection reagent is a one-step CRISPR nucleic acid detection reagent lyophilized body or a one-step Ago protein nucleic acid detection reagent lyophilized body.

Images