Component for reaction tube body and reaction tube

Abstract

The utility model relates to a component for a reaction tube body. The second aspect of the utility model relates to a reaction tube, in particular to a reaction tube suitable for two-step reaction, such as one-tube two-step method CRISPR (clustered regularly interspaced short palindromic repeats) nucleic acid detection. A component for a reaction tube body comprises a body, the body comprises a bottom face opening, a containing cavity and a side face opening, the containing cavity is communicated with the bottom face opening, and the side face opening is located in the side face of the body and communicated with the containing cavity; the limiting ring and the top of the body are of an integrated structure, and the size of the limiting ring is suitable for being clamped in a tube body of a reaction tube. The reaction tube comprises a tube body and a tube cover, and further comprises a part for the tube body of the reaction tube, which can be arranged in the tube body. The utility model solves the technical problem that the cover needs to be opened in two-step reaction, for example, the technical problem that the cover needs to be opened in two-step CRISPR (clustered regularly interspaced short palindromic repeats) nucleic acid detection.

Description

Technical Field

[0001] The first aspect of this utility model relates to a component for a reaction tube body; the second aspect of this utility model relates to a reaction tube, and more particularly to a reaction tube suitable for two-step reactions such as the one-tube two-step CRISPR nucleic acid detection. Background Technology

[0002] Reaction tubes are the most widely used consumables in molecular diagnostic technology applications. They are typically 0.2mL or 0.5mL capped conical tubes made of polypropylene or similar materials, and are commonly referred to as PCR tubes. PCR tubes are containers used for nucleic acid amplification and detection reactions such as qPCR, LAMP, RPA, and CRISPR. During the experiment, various reaction substances, including buffers, magnesium ions, primers, probes, nucleotides, enzymes, and DNA or RNA templates extracted from the sample, are precisely added using a pipette. The tubes are then sealed and placed in a nucleic acid amplification and detection instrument with matching tube types for temperature-controlled reaction. Simultaneously, fluorescence signals are detected to determine the presence of the target nucleic acid in the sample. Currently, these reaction tubes are mostly available in single-tube or eight-tube bundles and are widely used in research laboratories and professional clinical testing laboratories.

[0003] 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.

[0004] "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.

[0005] 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

[0006] The first objective of this invention is to provide a component for a reaction tube body to solve the technical problem that, in two-step experiments (such as two-step CRISPR nucleic acid detection), it is necessary to open the cap to add reagents required for the second step reaction after the first step reaction is completed.

[0007] The present invention solves the above-mentioned technical problems through the following technical solutions, thereby achieving the technical effects of the present invention.

[0008] A component for a reaction tube body, comprising:

[0009] The body includes a bottom opening, a receiving cavity, and a side opening. The receiving cavity communicates with the bottom opening, and the side opening is located on the side of the body and communicates with the receiving cavity.

[0010] A limiting ring is integrally formed with the top of the main body, and the size of the limiting ring is suitable for being fitted into the tube body of the reaction tube.

[0011] Preferably, based on any of the above technical solutions, the diameter of the bottom opening is 1-3mm, and the outer diameter of the limiting ring is 4-6mm.

[0012] Preferably, based on any of the above technical solutions, the outer diameter of the limiting ring matches the inner diameter of the upper part of the reaction tube.

[0013] Preferably, based on any of the above technical solutions, the capacity of the accommodating cavity is 0.5-20 μL. Preferably, based on any of the above technical solutions, the reaction tube body component is a reaction tube body component made of polypropylene.

[0014] Preferably, based on any of the above technical solutions, the component for the reaction tube body is white or transparent.

[0015] Preferably, based on any of the above technical solutions, the body is tubular, and the accommodating cavity and the bottom opening are integral structures.

[0016] Further, based on any of the above technical solutions, the component for the reaction tube body includes a protrusion located at the opening on the side of the body, the protrusion being open, and the opening of the protrusion communicating with the receiving cavity through the side opening.

[0017] Preferably, based on any of the above technical solutions, the body is 5-10mm long, and the inner diameter of the accommodating cavity is the same as the diameter of the opening on the bottom surface.

[0018] The second objective of this invention is to provide a reaction tube to solve the technical problem that in two-step experiments (such as two-step CRISPR nucleic acid detection), the tube needs to be opened to add reagents required for the second step reaction after the first step reaction is completed.

[0019] The present invention solves the above-mentioned technical problems through the following technical solutions, thereby achieving the technical effects of the present invention.

[0020] A reaction tube includes a tube body and a cap body, the cap body and the tube body being adapted to each other. The reaction tube also includes a reaction tube body component that can be disposed in the tube body, the reaction tube body component being as described in any of the first aspects of the technical solutions above.

[0021] The reaction tube components and reaction tubes of this invention are particularly suitable for use in a two-step method, such as in the two-step method of CRISPR nucleic acid detection, and in the detection of Argonaute protein (Ago enzyme) and nested PCR. Taking the two-step CRISPR nucleic acid detection method as an example, the beneficial effects of this invention are illustrated as follows: Before performing the first step of nucleic acid amplification reaction, the sample to be tested and nucleic acid amplification reaction reagent (a nucleic acid release reagent can also be added; the above reagents can also be added in lyophilized form or pre-filled in the reaction tube, and a reconstitution solvent is added when using) are added to the tube body of the reaction tube. The CRISPR nucleic acid detection reagent (the CRISPR nucleic acid detection reagent can also be added in lyophilized form or pre-filled, and a reconstitution solvent is added when using) is added to the tube body component, and the cap is closed. After the first step of nucleic acid amplification reaction is completed, without opening the cap a second time, the CRISPR nucleic acid detection reagent in the tube body component is allowed to enter the bottom of the reaction tube and mix with the nucleic acid amplification reaction product by centrifugation or hand shaking, and the second step of CRISPR nucleic acid detection reaction can then be carried out, 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

[0022] Figure 1 This is a front view structural diagram of the component for the reaction tube body according to a specific embodiment of this utility model.

[0023] Figure 2This is a schematic diagram of the reaction tube according to a specific embodiment of the present invention. Detailed Implementation

[0024] the term

[0025] 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.

[0026] The term "CRISPR" refers to clustered regularly interspaced short palindromic repeats, which originate from the immune system of microorganisms.

[0027] 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).

[0028] 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.

[0029] 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.

[0030] The term "Cas12a" (formerly "Cpf1") is a crRNA-dependent endonuclease, which is a type VA enzyme in the CRISPR system classification.

[0031] The term "Cas12b" (formerly "C2c1") is an sgRNA-dependent endonuclease, which is a type VB enzyme in the CRISPR system.

[0032] 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.

[0033] 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.

[0034] 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.

[0035] 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).

[0036] 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.

[0037] The term "temperature" refers to the temperature of the system (the mixture formed during testing).

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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).

[0043] 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).

[0044] In other embodiments, the biological sample may be plant cells, callus, tissue or organ (such as root, stem, leaf, flower, seed, fruit), etc.

[0045] 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.

[0046] 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.

[0047] The term "Ago protein" refers to Argonaute protein.

[0048] 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.

[0049] This specific embodiment proposes a component for the reaction tube body, and based on this, proposes product forms such as single tube and eight-tube array of reaction tubes.

[0050] See Figure 1 , 2 A component 1 for a reaction tube body includes: a body 11, a limiting ring 12, and a protrusion 13. The body 11 includes a bottom opening 110, a receiving cavity 111, and a side opening 112. The receiving cavity 111 communicates with the bottom opening 110, and the side opening 112 is located on the side of the body 11 and communicates with the receiving cavity 111. The diameter of the bottom opening 110 is 1-3 mm. The limiting ring 12 is integrally formed with the top of the body 11. The size of the limiting ring 12 is suitable for being fitted into the tube body 2 of the reaction tube, and the outer diameter of the limiting ring 12 is 4-6 mm. The outer diameter of the limiting ring 12 matches the upper inner diameter of the tube body 2 of the reaction tube, i.e., it is suitable for the limiting ring 12 to be fitted into the tube body 2 of the reaction tube. The capacity of the receiving cavity 111 is 2-20 μL. The component 1 for the reaction tube body is made of polypropylene. The component 1 for the reaction tube body is white or transparent. The main body 11 is tubular, with the accommodating cavity 111 and the bottom opening 110 being an integral structure. The reaction tube body component 1 includes a protrusion 13 located at the side opening 112 of the main body. The protrusion 13 is open, and its opening communicates with the accommodating cavity 111 through the side opening 112. The main body 11 is 5-10 mm long, and the inner diameter of the accommodating cavity 111 is the same as the diameter of the bottom opening 110.

[0051] A reaction tube includes a tube body 2 and a cap 3, and also includes a reaction tube body component 1 that can be disposed in the tube body, as described above.

[0052] Furthermore, the reaction tube also includes a lyophilized nucleic acid amplification reaction reagent 4, which is disposed within the tube body 2. When the lyophilized nucleic acid amplification reaction reagent 4 is pre-filled in the tube body 2, a sealing film can be applied to the opening of the tube body 2.

[0053] Furthermore, the reaction tube also includes a connecting part 5, through which the cover 3 is connected to the tube body 2. The reaction tube of this invention can also be made into an eight-unit row reaction tube. An eight-unit row reaction tube includes eight plastic transparent reaction tubes arranged in a row, wherein the plastic transparent reaction tubes are the reaction tubes described in any of the above technical solutions.

[0054] The reaction tube components and reaction tubes of this invention are particularly suitable for use in a two-step method, such as in the two-step method of CRISPR nucleic acid detection, and in the detection of Argonaute protein (Ago enzyme) and nested PCR. Taking the two-step CRISPR nucleic acid detection method as an example, the beneficial effects of this invention are illustrated as follows: Before performing the first step of nucleic acid amplification reaction, the sample to be tested and nucleic acid amplification reaction reagent (a nucleic acid release reagent can also be added) are added to the tube body 2 of the reaction tube; the above reagents can also be added in lyophilized form or pre-filled in the tube body of the reaction tube, and a reconstitution solvent is added when using. CRISPR nucleic acid detection reagent (CRISPR nucleic acid detection reagent can also be added in lyophilized form or pre-filled, and a reconstitution solvent is added when using) is added to the tube body component 1 of the reaction tube, and the cap 3 is closed. After the first step of nucleic acid amplification reaction is completed, without opening the cap a second time, the CRISPR nucleic acid detection reagent in the tube body component 1 of the reaction tube enters the bottom of the tube body 2 of the reaction tube and mixes with the nucleic acid amplification reaction product by centrifugation or hand shaking, and the second step of CRISPR nucleic acid detection reaction can be continued, 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 component for a reaction tube body, characterized in that, include: The body includes a bottom opening, a receiving cavity, and a side opening. The receiving cavity communicates with the bottom opening, and the side opening is located on the side of the body and communicates with the receiving cavity. A limiting ring is integrally formed with the top of the main body, and the size of the limiting ring is suitable for being fitted into the tube body of the reaction tube.

2. The component for the reaction tube body as described in claim 1, characterized in that, The diameter of the opening on the bottom surface is 1-3mm, and the outer diameter of the limiting ring is 4-6mm.

3. The component for the reaction tube body as described in claim 1, characterized in that, The outer diameter of the limiting ring matches the inner diameter of the upper part of the reaction tube.

4. The component for the reaction tube body as described in claim 1, characterized in that, The capacity of the accommodating cavity is 0.5-20 μL.

5. The component for the reaction tube body as described in claim 1, characterized in that, The components for the reaction tube body are made of polypropylene.

6. The component for the reaction tube body as described in claim 1, characterized in that, The components for the reaction tube body are white or transparent.

7. The component for the reaction tube body as described in claim 1, characterized in that, The main body is tubular, and the accommodating cavity and the bottom opening are integrally formed.

8. The component for the reaction tube body as described in claim 7, characterized in that, The component for the reaction tube body includes a protrusion located at an opening on the side of the body. The protrusion is open and communicates with the receiving cavity through the side opening.

9. The component for the reaction tube body as described in claim 8, characterized in that, The body is 5-10mm long, and the inner diameter of the accommodating cavity is the same as the diameter of the opening on the bottom surface.

10. A reaction tube, comprising a tube body and a cap, wherein the cap and the tube body are adapted to fit together, characterized in that, The reaction tube further includes a reaction tube body component that can be disposed in the tube body, the reaction tube body component being as described in any one of claims 1-9.

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