CIRSPR nucleic acid detection reagent freeze-dried body and preloaded product thereof

Abstract

The utility model relates to a detection reagent, in particular to a CIRSPR nucleic acid detection reagent. The CIRSPR nucleic acid detection reagent freeze-dried body comprises a body, the body is a particle prepared from CIRSPR nucleic acid detection reagent freeze-dried powder, the CIRSPR nucleic acid detection reagent freeze-dried body further comprises an inert fragile material wrapping layer, and the inert fragile material wrapping layer wraps the body. The utility model relates to a preloaded product of a CIRSPR nucleic acid detection reagent. The preloaded product comprises a CIRSPR nucleic acid detection reagent freeze-dried body and a preloaded container, according to the two-step CRISPR nucleic acid detection device, the technical problem that a cover needs to be opened to add a reagent required by a second-step reaction after the first-step reaction is completed in two-step CRISPR nucleic acid detection is solved.

Description

Technical Field

[0001] This utility model relates to a detection reagent, and more particularly to a CIRSPR nucleic acid detection reagent. 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 CRISPR nucleic acid detection, it is necessary to open the container and add the reagents required for the second step reaction after completing the first step reaction. Utility Model Content

[0005] The first objective of this invention is to provide a lyophilized CRISPR nucleic acid detection reagent to solve the technical problem that the reagents required for the second step reaction need to be added after the first step reaction in the two-step CRISPR nucleic acid detection method.

[0006] The present invention solves the above-mentioned technical problems through the following technical solutions, thereby achieving the purpose of the present invention.

[0007] A CIRSPR nucleic acid detection reagent lyophilized body includes a main body, wherein the main body is a particle made of CIRSPR nucleic acid detection reagent lyophilized powder, and the CIRSPR nucleic acid detection reagent lyophilized body further includes an inert and fragile material coating layer, wherein the inert and fragile material coating layer coats the main body.

[0008] Preferably, the body is spherical with a diameter of 1-5 mm.

[0009] Preferably, the thickness of the inert and fragile material coating layer is 0.5-2 mm.

[0010] Preferably, the inert and fragile material used for the inert and fragile material coating layer is one or more of paraffin wax, rosin, polyethylene wax, palm wax, and ceresin. This inert and fragile material does not participate in nucleic acid amplification or CRISPR nucleic acid detection reactions, and is easily crushed, exposing the CRISPR nucleic acid detection reagents.

[0011] The second objective of this invention is to provide a pre-packaged CRISPR nucleic acid detection reagent product to solve the technical problem that the two-step CRISPR nucleic acid detection method requires opening the cap to add reagents for the second step reaction after the first step reaction is completed.

[0012] The present invention solves the above-mentioned technical problems through the following technical solutions, thereby achieving the purpose of the present invention.

[0013] A pre-packaged CIRSPR nucleic acid detection reagent product includes:

[0014] CIRSPR nucleic acid detection reagent lyophilized body, wherein the CIRSPR nucleic acid detection reagent lyophilized body is as described in any of the technical solutions of the first aspect above;

[0015] A pre-filled container, which is a pre-filled reaction tube or a pre-filled plate, wherein the pre-filled reaction tube includes a cap, a sealing film, and a flexible tube body, and the CIRSPR nucleic acid detection reagent lyophilized body is pre-filled in the flexible tube body, the sealing film seals the opening of the flexible tube body, and the cap body is adapted to the flexible tube body; the pre-filled plate includes a cover plate, a sealing film, a flexible tube body assembly, and a connecting plate, wherein the flexible tube body assembly includes a plurality of flexible tube bodies, the CIRSPR nucleic acid detection reagent lyophilized body is pre-filled in each of the flexible tube bodies, the connecting plate connects the plurality of flexible tube bodies, the sealing film seals the opening of the plurality of flexible tube bodies, and the cover plate is adapted to the flexible tube body assembly.

[0016] Preferably, the flexible tube is a white or transparent conical flexible tube.

[0017] Furthermore, the pre-assembled reaction tube also includes a connecting part, which connects the cover and the flexible tube.

[0018] Preferably, the pre-assembled plate is a 6-hole plate or a 12-hole plate.

[0019] This invention relates to a CRISPR nucleic acid detection reagent lyophilized body and its pre-packaged product, which has a simple structure and is easy to use, especially suitable for use in two-step methods. Taking the two-step CRISPR nucleic acid detection as an example, the beneficial effects of this invention are illustrated as follows: Before the nucleic acid amplification reaction, the sample to be tested, nucleic acid amplification reaction reagent, and reconstitution solvent (a nucleic acid release reagent can also be added) are added to the flexible tube; the above reagents can also be pre-packaged in lyophilized powder form, and the sample to be tested and reconstitution solvent are added when using. Then the cap is closed. After the nucleic acid amplification reaction is completed, without opening the cap a second time, the lyophilized CRISPR nucleic acid detection reagent can be crushed by squeezing the flexible tube by hand to continue the second step of the CRISPR nucleic acid detection reaction, preventing contamination (such as aerosol contamination) caused by opening the cap a second time and the resulting problems such as false positives. Attached Figure Description

[0020] Figure 1 This is a cross-sectional view of the lyophilized form of the CIRSPR nucleic acid detection reagent according to a specific embodiment of this utility model.

[0021] Figure 2 This is a schematic diagram of the pre-loaded reaction tube according to a specific embodiment of the present invention.

[0022] Figure 3 This is a structural schematic diagram of a specific embodiment of the present invention. Detailed Implementation

[0023] the term

[0024] 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.

[0025] The term "CRISPR" refers to clustered regularly interspaced short palindromic repeats, which originate from the immune system of microorganisms.

[0026] 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).

[0027] 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.

[0028] 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.

[0029] The term "Cas12a" (formerly "Cpf1") is a crRNA-dependent endonuclease, which is a type VA enzyme in the CRISPR system classification.

[0030] The term "Cas12b" (formerly "C2c1") is an sgRNA-dependent endonuclease, which is a type VB enzyme in the CRISPR system.

[0031] 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.

[0032] 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.

[0033] 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.

[0034] 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 CN 110551800 A, with an application publication date of December 10, 2019, first disclosed a one-step method (see paragraphs

[0238] ,

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

[0035] 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.

[0036] The term "temperature" refers to the temperature of the system (the mixture formed during testing).

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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, Smith and 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 computerized operation of 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.

[0041] The term "nucleic acid analogue" 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).

[0042] 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).

[0043] In other embodiments, the biological sample may be plant cells, callus, tissue or organ (such as root, stem, leaf, flower, seed, fruit), etc.

[0044] The "sample to be tested" may contain 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. The amplification technique is isothermal amplification, which 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 (second primer isothermal amplification). The following are some of the amplification methods: intelligent amplification method, 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 induced amplification), WGA (whole genome amplification), PSR (polymerase helical reaction), or combinations thereof.

[0045] Furthermore, the detection method of this 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.

[0046] See Figure 1A lyophilized CRISPR nucleic acid detection reagent 1 comprises a main body 11 and an inert and fragile material coating layer 12. The main body 11 is a granule made from the lyophilized CRISPR nucleic acid detection reagent powder 11, and is spherical with a diameter of 1-5 mm. The inert and fragile material coating layer 12 encapsulates the main body 11, and the thickness of the inert and fragile material coating layer 12 is 0.5-2 mm. The inert and fragile material used in the inert and fragile material coating layer 12 is one or more of paraffin wax, rosin, polyethylene wax, palm wax, and ceresin. This inert and fragile material does not participate in nucleic acid amplification or the CRISPR nucleic acid detection reaction, and is easily crushed, exposing the CRISPR nucleic acid detection reagent.

[0047] A pre-packaged CIRSPR nucleic acid detection reagent product includes: CIRSPR nucleic acid detection reagent lyophilized 1 and a pre-packaged container.

[0048] As described above, the CIRSPR nucleic acid detection reagent lyophilized body 1 comprises a main body 11 and an inert and fragile material coating layer 12. The main body 11 is a granule made from the CIRSPR nucleic acid detection reagent lyophilized powder, and is spherical with a diameter of 1-5 mm. The inert and fragile material coating layer 12 encapsulates the main body 11, and the thickness of the inert and fragile material coating layer 12 is 0.5-2 mm. The inert and fragile material used in the coating layer is one or more of paraffin wax, rosin, polyethylene wax, palm wax, and ceresin. This inert and fragile material does not participate in nucleic acid amplification or the CRISPR nucleic acid detection reaction, and is easily crushed, exposing the CIRSPR nucleic acid detection reagent.

[0049] See Figure 2 In one embodiment, the pre-filled container is a pre-filled reaction tube 2, which includes a cap 21, a flexible tube 22, and a sealing film (not shown in the figure), and may further include a connecting part 23. The CIRSPR nucleic acid detection reagent lyophilized body 1 is pre-filled in the flexible tube 22. The sealing film seals the opening of the flexible tube 22; the sealing film is removed before use. The cap 21 is adapted to the flexible tube 22 and is free from the flexible tube 22. It can be connected to the flexible tube 22 via the connecting part 23. Before use, the sample to be tested, the nucleic acid amplification reaction reagent, and the reconstitution solvent are added, and then the cap 21 is closed. The flexible tube 22 is a white or transparent conical flexible tube.

[0050] See Figure 3In another embodiment, the pre-filled container is a pre-filled plate 3; the pre-filled plate 3 includes a cover plate 31, a sealing film (not shown in the figure), a flexible tube assembly 32, and a connecting plate 33. The flexible tube assembly 32 includes a plurality of flexible tubes 321, with the CIRSPR nucleic acid detection reagent lyophilized body 1 pre-filled in each flexible tube 321. The connecting plate 33 connects the plurality of flexible tubes 321, and the sealing film seals the openings of the plurality of flexible tubes 321; the sealing film is removed before use. The cover plate 31 is adapted to the flexible tube assembly 32 and is free from the flexible tubes 321; the cover plate 31 is closed after adding the sample to be tested, the nucleic acid amplification reaction reagent, and the reconstitution solvent. The flexible tubes 321 are white or transparent conical flexible tubes. The pre-filled plate is a 6-well plate or a 12-well plate.

[0051] This invention relates to a CRISPR nucleic acid detection reagent lyophilized body and its pre-packaged product, which has a simple structure and is easy to use, especially suitable for use in a two-step method. Taking the two-step method of CRISPR nucleic acid detection as an example, the beneficial effects of this invention are illustrated as follows: Before the nucleic acid amplification reaction, the sample to be tested, nucleic acid amplification reaction reagent, and reconstitution solvent (a nucleic acid release reagent can also be added) are added to the flexible tube; the above reagents can also be pre-packaged in lyophilized powder form, and the sample to be tested and reconstitution solvent are added during use. Then, the cap is closed. After the nucleic acid amplification reaction is completed, without opening the cap a second time, the lyophilized CRISPR nucleic acid detection reagent can be crushed by squeezing the flexible tube by hand to continue the second step of the CRISPR nucleic acid detection reaction, preventing contamination (such as aerosol contamination) caused by opening the cap a second time and the resulting problems such as false positives.

Claims

1. A CIRSPR nucleic acid detection reagent lyophilized body, comprising a bulk, wherein the bulk is granules made from CIRSPR nucleic acid detection reagent lyophilized powder, characterized in that, The CIRSPR nucleic acid detection reagent lyophilized body further includes an inert and fragile material coating layer, which encapsulates the main body.

2. The CIRSPR nucleic acid detection reagent lyophilized form as described in claim 1, characterized in that, The body is spherical with a diameter of 1-5 mm.

3. The CIRSPR nucleic acid detection reagent lyophilized form as described in claim 1, characterized in that, The thickness of the inert and fragile material coating layer is 0.5-2 mm.

4. A pre-packaged CIRSPR nucleic acid detection reagent product, characterized in that, include: The CIRSPR nucleic acid detection reagent lyophilized form is as described in any one of claims 1-3; A pre-filled container, which is a pre-filled reaction tube or a pre-filled plate, wherein the pre-filled reaction tube includes a cap, a sealing film, and a flexible tube body, and the CIRSPR nucleic acid detection reagent lyophilized body is pre-filled in the flexible tube body, the sealing film seals the opening of the flexible tube body, and the cap body is adapted to the flexible tube body; the pre-filled plate includes a cover plate, a sealing film, a flexible tube body assembly, and a connecting plate, wherein the flexible tube body assembly includes a plurality of flexible tube bodies, the CIRSPR nucleic acid detection reagent lyophilized body is pre-filled in each of the flexible tube bodies, the connecting plate connects the plurality of flexible tube bodies, the sealing film seals the opening of the plurality of flexible tube bodies, and the cover plate is adapted to the flexible tube body assembly.

5. A pre-packaged CIRSPR nucleic acid detection reagent product as described in claim 4, characterized in that, The flexible tube is a white or transparent conical flexible tube.

6. The CIRSPR nucleic acid detection reagent pre-packaged product as described in claim 4, characterized in that, The pre-installed reaction tube further includes a connecting part that connects the cover and the flexible tube.

7. A pre-packaged CIRSPR nucleic acid detection reagent product as described in claim 4, characterized in that, The pre-assembled plate is a 6-hole plate or a 12-hole plate.

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