Component for reaction tube cap body, cap body and reaction tube

Abstract

The utility model relates to a component for a reaction tube cap body. The utility model further relates to a reaction tube cap body. The utility model further relates to a reaction tube on the third aspect, in particular to a reaction tube suitable for two-step reaction such as one-tube two-step method CRI SPR nucleic acid detection. The component for the reaction tube cap body is characterized in that the component for the reaction tube cap body comprises a body, the body is in a hollow cone shape and comprises an upper end opening and a lower end opening hole, and the size of the body is suitable for being clamped in a cap body cavity of the reaction tube cap body. The utility model solves the technical problem that after the first-step reaction of two-step experiments (such as a two-step method of CRI SPR detection) is completed, a cover needs to be opened to add a reagent required by the second-step reaction.

Description

TECHNICAL FIELD

[0001] The first aspect of the utility model relates to a kind of components for reaction tube cap body;The second aspect of the utility model relates to a kind of reaction tube cap body;The third aspect of the utility model further relates to a kind of reaction tube, especially relates to a kind of reaction tube suitable for two-step reaction such as one pipe two-step method CRISPR nucleic acid detection. BACKGROUND

[0002] Reaction tube is the most widely used consumable in molecular diagnostic technology application, generally 0.2mL or 0.5mL of conical tube with cover, made of polypropylene or similar material, usually referred to as PCR tube. PCR tube is the container for nucleic acid amplification and detection reaction such as qPCR, LAMP, RPA, CRISPR etc. In the experiment process, pipette is generally used to accurately add various reaction materials, including buffer, magnesium ion, primer, probe, nucleotide, enzyme, DNA or RNA template extracted in sample etc., then cover is sealed and placed in nucleic acid amplification detector of matching tube type to carry out temperature control reaction, and fluorescence signal for judging whether the nucleic acid to be detected in sample exists is detected. At present, this kind of reaction tube exists in the form of single tube or eight-tube product, and is mostly used in scientific research laboratory and professional clinical laboratory.

[0003] CRISPR detection, including CRISPR nucleic acid detection and CRISPR non-nucleic acid detection. CRISPR nucleic acid detection is the abbreviation of CRISPR / Cas system nucleic acid detection. CRISPR nucleic acid detection is a nucleic acid detection based on the system developed from CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and Cas protein (abbreviation of CRISPR associated protein). CRISPR nucleic acid detection includes nucleic acid detection using the cis cleavage activity of Cas protein and nucleic acid detection using the trans cleavage activity of Cas protein. The components required for CRISPR nucleic acid detection using the trans cleavage activity of Cas protein include Cas protein, guide RNA and single-stranded nucleic acid and / or nucleic acid analogue reporter molecule (also referred to as nucleic acid probe in some documents). The core of this nucleic acid detection is still the principle of base pairing, i.e. the guide sequence of guide RNA is paired with target nucleic acid, and Cas protein acts to bind with the direct repeat sequence (DR) of guide RNA, and after the guide sequence of guide RNA is paired with target nucleic acid, it can be activated to cut activity. The activation of cutting activity is used by people to carry out signal reporting to obtain qualitative or quantitative information of target nucleic acid detection. Therefore, Cas protein can also be regarded as part of signal reporting.

[0004] The "one-step" refers to that the nucleic acid amplification and the CRISPR nucleic acid detection are implemented in one step. The concept similar to the "one-step" is "one-pot" (also referred to as "one-pot method"). The "one-pot" refers to that the nucleic acid amplification and the CRISPR nucleic acid detection are implemented in one container, which can be a two-step method or a one-step method.

[0005] Currently, when the two-step reaction such as the two-step method of the CRISPR nucleic acid detection is carried out, the reagent required for the second step reaction needs to be added after the first step reaction is completed. Practical new content

[0006] The first aspect of the utility model is to provide a component for a reaction tube cap body, so as to solve the technical problem that the two-step experiment (such as the two-step method of the CRISPR nucleic acid detection) needs to add the reagent required for the second step reaction after the first step reaction is completed.

[0007] The utility model solves the above technical problem and achieves the technical effect of the utility model by the following technical scheme.

[0008] A component for a reaction tube cap body, the component for the reaction tube cap body comprises a body, the body is in a hollow conical shape or a hollow arc shape, comprises an upper end opening and a lower end opening, the size of the body is suitable for being clamped in the cap body cavity of the reaction tube cap body, and the aperture of the lower end opening is 1-3mm.

[0009] Preferably, the outer diameter of the upper end of the body is 4-6mm.

[0010] Preferably, the outer diameter of the upper end of the body matches the inner diameter of the cap body cavity of the reaction tube cap body. Preferably, the capacity of the component for the reaction tube cap body is 0.5-20μL.

[0011] Preferably, the component for the reaction tube cap body is a component for the reaction tube cap body made of polypropylene. Preferably, the component for the reaction tube cap body is a white or transparent component for the reaction tube cap body. Further, the component for the reaction tube cap body further comprises a through-hole tube body, the upper end of the through-hole tube body is in an integral structure with the lower end of the body, the through-hole tube body has a through hole penetrating through the upper end and the lower end, and the through-hole tube body is in communication with the lower end opening of the body. Preferably, the length of the through-hole tube body is 5-10mm, and the inner diameter of the through-hole tube body is the same as the aperture of the lower end opening.

[0012] The second aspect of the utility model is to provide a reaction tube cap body, so as to solve the technical problem that the two-step experiment (such as the two-step method of the CRISPR nucleic acid detection) needs to add the reagent required for the second step reaction after the first step reaction is completed.

[0013] The utility model discloses the following technical scheme solves above-mentioned technical problem, achieves the technical effect of the utility model.

[0014] A reaction tube cap body, comprising a reaction tube cap body body, the reaction tube cap body body comprising a cap body cavity, further comprising a reaction tube cap body component;The reaction tube cap body component can be clamped in the cap body cavity, or the reaction tube cap body component is arranged in the cap body cavity and is integrated, and the reaction tube cap body component is as any one of the first aspect described above.

[0015] The third aspect of the utility model is to provide a reaction tube to solve the technical problem that two-step experiments (such as the two-step method of CRISPR detection) need to open the cover to add reagents required for the second step reaction after completing the first step reaction.

[0016] The utility model discloses the following technical scheme solves above-mentioned technical problem, achieves the technical effect of the utility model.

[0017] A reaction tube, comprising a reaction tube cap body and a reaction tube body, the reaction tube cap body is matched with the reaction tube body, and the reaction tube cap body is as any one of the second aspect described above.

[0018] The reaction tube cap body component, the reaction tube cap body and the reaction tube of the utility model are especially suitable for use in one-tube two-step method, such as use in one-tube two-step method of CRISPR nucleic acid detection, and also such as use in Argonaute protein (Ago enzyme) detection and nested PCR.Taking one-tube two-step method of CRISPR detection as an example, the beneficial effects of the utility model are as follows: when carrying out the first step nucleic acid amplification reaction, adding the sample to be detected and nucleic acid amplification reagent (such as also adding nucleic acid release reagent) in the reaction tube body, adding CRISPR nucleic acid detection reagent in the reaction tube cap body component, and covering the cap body, after the first step nucleic acid amplification reaction is finished, without opening the cover again, the CRISPR detection nucleic acid reagent in the reaction tube cap body component enters the bottom of the reaction tube body and mixes with the nucleic acid amplification reaction product by centrifugation or hand shaking, and the second step CRISPR nucleic acid detection reaction can be continued, preventing pollution (such as aerosol pollution) caused by opening the cover again and problems such as false positive caused thereby. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 It is the front view structural schematic diagram of the detection component for the reaction tube of the embodiment of the utility model.

[0020] Figure 2 It is the top view structural schematic diagram of the detection component for the reaction tube of the embodiment of the utility model.

[0021] Figure 3 is a structural schematic diagram of the reaction tube of the embodiment of the present application. DETAILED DESCRIPTION

[0022] TERMS

[0023] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0024] The term "CRISPR" refers to Clustered Regularly Interspaced Short Palindromic Repeats, which is from the immune system of microorganisms.

[0025] The term "CRISPR-Cas": a unique genomic element derived from bacteria and archaea, as an adaptive immune defense system to resist invading phages or foreign nucleic acids. The system is composed of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated proteins (abbreviated as Cas proteins, Cas).

[0026] The term "Cas protein" refers to CRISPR-associated protein, which is a related protein in the CRISPR system. The "Cas protein" described herein refers to CRISPR-associated protein (some literature translates it as CRISPR-Cas effector protein, CRISPR / Cas effector protein, CRISPR-Cas effector, CRISPR / Cas effector). The Cas proteins currently detected include type I Cas protein (Cas3), type II Cas protein (Cas9), type III Cas protein (Cas10), type V Cas protein (Cas12) or type VI Cas protein (Cas13). In particular, type V Cas protein (Cas12), type VI Cas protein (Cas13) and part of Cas3, Cas10 are found to have trans-cleavage activity, which can realize signal amplification for detection, so the trans-cleavage activity is often used for detection. Taking type V Cas protein as an example, once it binds to the cis-cleavage substrate under the guidance of the guide RNA to form a ternary complex of Cas protein-guide RNA-cis-cleavage substrate, it can induce its trans-cleavage activity, i.e. random cleavage of single-stranded DNA (including base-modified single-stranded DNA), and there are also reports that it will randomly cleave single-stranded nucleic acid analogs. Of course, the cis-cleavage activity of Cas protein or other properties can also be used for detection.

[0027] The Cas protein described in the present embodiment is preferably a protein having trans-cleavage activity. In particular, the Cas protein is active, especially trans-cleavage active, at a temperature higher than the temperature of the system in which the isothermal amplification reaction is performed.

[0028] The term "Cas12a" (formerly "Cpf1") is a crRNA-dependent endonuclease, which is a V-A type enzyme in the classification of CRISPR systems.

[0029] The term "Cas12b" (formerly "C2c1") is a sgRNA-dependent endonuclease, which is a V-B type enzyme in the classification of CRISPR systems.

[0030] The term "PAM" refers to a protospacer-adjacent motif, which is a short DNA sequence directly adjacent to the DNA sequence targeted by the CRISPR effector protein, and is necessary for Cas12a or Cas12b to cleave double-stranded DNA, for example, the PAM of Cas12a is TTTV, and the PAM of AacCas12b is a TTN sequence.

[0031] The term "target DNA or RNA molecule" refers to a DNA or RNA or a specific part thereof to be tested when the nucleic acid molecule to be tested is a nucleic acid molecule; when the non-nucleic acid molecule to be tested is a nucleic acid sequence designed in advance.

[0032] The term "CRISPR nucleic acid detection method" refers to a nucleic acid detection method using a Cas protein, including a nucleic acid detection method using the Cas protein for cis-cleavage activity, trans-cleavage activity or other functions.

[0033] The term "one-step CRISPR nucleic acid detection method (using Cas protein transcleavage activity)" (or simply CRISPR one-step nucleic acid detection, CRISPR one-step, one-step detection, one-step) is a rapid and convenient detection technology developed on the basis of the CRISPR nucleic acid detection system, which can simultaneously realize the amplification and detection of target nucleic acid in one reaction tube. This technology combines CRISPR-Cas system and isothermal amplification (or constant temperature amplification) technology, and does not need to open the cover operation after amplification of nucleic acid product, which can specifically detect the target nucleic acid in a short time. The CRISPR one-step detection technology is a rapid, accurate, high sensitivity and high specificity detection technology, which is not only simple to operate, but also can improve the detection specificity of the current isothermal amplification technology. Compared with the traditional PCR technology, the CRISPR one-step detection does not need complex temperature control and multi-step operation, and has higher real-time and portability. The Chinese invention patent with application publication number CN 110551800 A and application publication date 2019.12.10 first discloses one-step method (see

[0238] ,

[0239] and other paragraphs of the patent application).

[0034] The term "system" should be understood broadly, which can be a composition, a product combination, a reagent, a kit, an instrument containing the aforementioned composition, product combination, reagent, kit, a mixture (system) formed when the composition, product combination, reagent, kit is used for detection, and an instrument containing the aforementioned mixture, etc.

[0035] The term "temperature" refers to the temperature of the system (mixture formed when used for detection).

[0036] The term "guide RNA" is a mature crRNA fused (or not fused) with tracrRNA as a guide RNA, or a mature crRNA fused (or not fused) with scoutRNA as a guide RNA, or a crRNA alone as a guide RNA.

[0037] Generally, a guide RNA (gRNA) can comprise, consist essentially of, or consist of direct repeat sequences (also referred to as DR sequences) and a guide sequence. The gRNA can comprise, consist essentially of, or consist of crRNA and tracrRNA, crRNA and scoutRNA, or only crRNA, depending on the Cas protein it relies on in different Type V CRISPR systems. The crRNA and tracrRNA can be artificially engineered to form a single guide RNA (sgRNA). In some cases, the guide sequence is a polynucleotide sequence that is of sufficient complementarity to hybridize to and direct specific binding of a CRISPR / Cas protein-guide RNA complex to a cis-cleavage substrate nucleic acid, typically having a sequence length of 15-28 nt in Type V CRISPR systems. The direct repeat sequences can fold to form a specific structure (e.g., stem-loop structure) for recognition by the Cas protein to form a complex. The guide sequence does not need to be 100% complementary to the cis-cleavage substrate nucleic acid. The guide sequence is not complementary to the nucleic acid in the trans-cleavage reporter molecule.

[0038] In some embodiments, the degree of complementarity (match) between the guide sequence and its corresponding cis-cleavage substrate nucleic acid, when optimally aligned, is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%. Determining optimal alignment is within the capabilities of a person of ordinary skill in the art. For example, there are publicly available and commercially available alignment algorithms and programs, such as, but not limited to, ClustalW, Smith-Waterman in matlab, Bowtie, Geneious, Biopython, and SeqMan. The terms “polynucleotide,” “nucleotide sequence,” “nucleic acid sequence,” “nucleic acid molecule,” and “nucleic acid” can be used interchangeably and include DNA, RNA, or a hybrid thereof, which can be double-stranded or single-stranded, unless indicated otherwise.

[0039] The terms "homology" or "identity" are used in reference to the matching of sequences between two polypeptides or between two nucleic acids. When a position in each of two sequences being compared is occupied by the same base or amino acid monomer subunit (e.g., a position in each of two DNA molecules occupied by adenine, or a position in each of two polypeptides occupied by lysine), then the molecules are identical at that position. Generally, comparisons are made using the complete sequences aligning the two sequences to produce the maximum identity. Such alignments can be determined, for example, by using computerized running of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group) which are routinely used and are available, e.g., using the default parameters, through the University of Wisconsin Biotechnology Center. BLAST algorithms, available from the National Center for Biotechnology Information (NCBI www.ncbi.nlm.nih.gov / ), can also be used.

[0040] The term "nucleic acid analogues" is a class of derivatives of RNA and DNA, which is mainly composed of phosphate, pentose and base, while the nucleic acid analogue replaces at least one of them with some other substance. The main nucleic acid analogues are peptide nucleic acid (PNA), morpholino (MNA), bridged nucleic acid (BNA), locked nucleic acid (LNA), glycol nucleic acid (GNA) and threose nucleic acid (TNA). Some of these nucleic acid analogues can even carry out biological processes such as replication, translation, etc. in vitro (Brudno, Yevgeny; Birnbaum, Michael E; Kleiner, Ralph E; Liu, David R. "An in vitro translation, selection and amplification system for peptide nucleic acids". Nature Chemical Biology. 6 (2): 148-155. doi:10.1038 / nchembio.280. PMC 2808706. PMID 20081830).

[0041] The term "test sample" refers to a sample obtained from a biological sample that has been subjected to extraction of nucleic acids, which sample can also have been subjected to amplification, transcription, reverse transcription of nucleic acids. The biological sample is any solid or fluid sample obtained, excreted or secreted from any organism, including but not limited to unicellular organisms, such as bacteria, yeast, protozoa and amoebae, and multicellular organisms, such as plants or animals, including samples from a healthy or apparently healthy human subject or a human patient affected by a condition or disease to be diagnosed or investigated, for example, an infection with a pathogenic microorganism, such as a pathogenic bacterium or virus. For example, the 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 body secretion, exudate, exudate (e.g., fluid obtained from an abscess or any other site of infection or inflammation), or a swab of a skin or mucosal surface, or a fluid obtained from a joint (e.g., a normal joint or a joint affected by a disease, such as rheumatoid arthritis, osteoarthritis, gout or septic arthritis). The sample can also be a sample obtained from any organ or tissue (including a biopsy or autopsy specimen, such as a tumor biopsy) or can comprise cells (primary cells or cultured cells) or a medium conditioned by any cell, tissue or organ. Exemplary samples include, but are not limited to, cells, cell lysates, blood smears, 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).

[0042] In other embodiments, the biological sample can be a plant cell, callus, tissue or organ (e.g., roots, stems, leaves, flowers, seeds, fruits), etc.

[0043] The "sample to be tested" can contain a nucleic acid molecule to be tested. In the present application, the nucleic acid molecule to be tested includes a DNA molecule, and also includes an RNA molecule or a DNA molecule formed by reverse transcription of an RNA molecule, or further, the nucleic acid molecule to be tested can be amplified by a technique known in the art, and the amplification technique is an isothermal amplification technique, and the isothermal amplification can be LAMP (loop-mediated isothermal amplification), RPA (recombinase polymerase amplification), RAA (recombinase-mediated amplification), ERA (enzyme recombination isothermal amplification technique), MIRA (multi-enzyme isothermal rapid amplification technique), bDNA (branch DNA amplification), NASBA (nucleic acid sequence-based amplification), SDA (strand displacement amplification), TMA (transcription-mediated amplification), RCA (rolling circle amplification), HDA (helicase-dependent amplification), SPIA (single primer isothermal amplification), NEAR (nicking enzyme amplification reaction), SMAP (smart amplification process), SMAP2 (second edition of smart amplification process), CPA (crossover primer amplification), MDA (multiple displacement amplification), RAM (Ramification), cHDA (helicase-dependent circular amplification), SMART (signal-mediated amplification of RNA technique), 3SR (self-sustained sequence replication system), GEAR (genomic exponential amplification reaction), IMDA (isothermal multiple displacement amplification), ERA (enzyme recombination isothermal amplification), TAS (transcription-dependent amplification system), RIDA (rapid isothermal detection amplification technique), NEMA (nicking endonuclease isothermal amplification of nucleic acids), EXPAR (exponential isothermal amplification), ICAN (isothermal chimeric primer-initiated amplification of nucleic acids), SEA (strand exchange amplification), SHARP (SSB-helicase-mediated rapid PCR), IMSA (isothermal multiple self-priming amplification), WGA (whole genome amplification), PSR (polymerase spiral reaction), or a combination thereof.

[0044] Further, the detection method of the present application further comprises a step of amplifying the nucleic acid molecule to be tested; and the detection system further comprises components for amplifying the nucleic acid molecule to be tested. The components for amplification include one or more of the following: a DNA polymerase, a reverse transcriptase, a strand displacement enzyme, a nicking endonuclease, a helicase, a recombinase, a single-strand binding protein, a recombination modulating protein, a T7 RNA polymerase, an RNase H, dNTPs for amplification reaction and / or reverse transcription reaction, NTPs for transcription reaction, a buffer, and the like.

[0045] The term "Ago protein" is an Argonaute protein.

[0046] The term "Ago protein nucleic acid detection" refers to nucleic acid detection using Argonaute protein, such as "Nucleic acid detection method based on prokaryotic Argonaute protein and its application" disclosed in Chinese invention patent CN108796036A, "Nucleic acid detection method based on normal temperature prokaryotic Argonaute protein and its application" disclosed in CN114277109A, "Visual detection system, reagent or kit for detecting target nucleic acid molecules and detection method" disclosed in CN114085892A, and "Nucleic acid detection method based on medium temperature Argonaute protein and isothermal amplification" disclosed in CN116064736A. In addition, short pAgo and its related nuclease effect protein can form a heterodimer complex (TmuRE-Ago complex); unlike long pAgo which specifically cuts target DNA, this complex is activated after RNA-guided DNA target recognition, exhibiting high non-specific DNA cleavage activity (see https: / / doi.org / 10.1093 / nar / gkad1145). This non-specific DNA cleavage activity can also be used for detection.

[0047] Referring to Figure 1 、 2 A reaction tube cap component 1 includes a body 11 and can also include a through-hole tube 12. The body 11 is hollow conical or hollow arc-shaped, includes an upper end opening 110 and a lower end opening 111, and the size of the body 11 is suitable for being clamped in the cap cavity (not shown in the figure) of the reaction tube cap 2, and the aperture of the lower end opening is 1-3 mm.

[0048] The upper end outer diameter of the body 11 is 4-6 mm. The upper end outer diameter of the body 11 matches the inner diameter of the cap cavity of the reaction tube cap 2. The capacity of the reaction tube cap component 1 is 0.5-20 mL. The reaction tube cap component 1 is a reaction tube cap component made of polypropylene. The reaction tube cap component 1 is a white or transparent reaction tube cap component. The upper end of the through-hole tube 12 is in an integral structure with the lower end of the body 11, the through-hole tube 12 has a through-hole penetrating through the upper end and the lower end, and the through-hole tube 12 communicates with the lower end opening of the body 11. The length of the through-hole tube is 5-10 mm, and the inner diameter of the through-hole tube 12 is the same as the aperture of the lower end opening.

[0049] A reaction tube cap 2 includes a reaction tube cap body 21, which includes a cap cavity and a reaction tube cap component 1; the reaction tube cap component 1 can be clamped in the cap cavity, or the reaction tube cap component 1 is arranged in the cap cavity and in an integral structure, and the reaction tube cap component 1 is as described in any one of the first aspect.

[0050] Referring to Figure 3A reaction tube comprises a reaction tube cap 2 and a reaction tube body 3, the reaction tube cap 2 is matched with the reaction tube body 3, and the reaction tube cap 2 is as described in any of the technical solutions of the second aspect.

[0051] Further, the reaction tube further comprises a nucleic acid amplification reaction reagent freeze-dried body 4 arranged in the reaction tube body. The reaction tube further comprises a connecting part 5, and the reaction tube cap 2 is connected to the reaction tube body 4 through the connecting part 5.

[0052] The reaction tube of the utility model can also be made into eight-row reaction tubes. An eight-row reaction tube comprises eight plastic transparent reaction tubes arranged in a row, and the plastic transparent reaction tube is the reaction tube as described in any of the technical solutions.

[0053] The reaction tube cap, the reaction tube cap part and the reaction tube of the utility model are especially suitable for use in one-tube two-step method, such as use in one-tube two-step method of CRISPR nucleic acid detection, and use in Argonaute protein (Ago enzyme) detection and nested PCR. The beneficial effects of the utility model are described below by taking the one-tube two-step method of CRISPR detection as an example: when performing the first step of nucleic acid amplification reaction, the sample to be detected and the nucleic acid amplification reaction reagent (such as a nucleic acid release reagent; the reagent can also be made into a freeze-dried body and preloaded with a reconstitution agent) are added to the reaction tube body, the CRISPR nucleic acid detection reagent is added to the reaction tube cap part, and the cap is covered. After the first step of nucleic acid amplification reaction is completed, without opening the cap again, the CRISPR nucleic acid detection reagent in the reaction tube cap part is mixed with the nucleic acid amplification reaction product at the bottom of the reaction tube body by centrifugation or hand shaking, and the second step of CRISPR nucleic acid detection reaction can be continued, thereby preventing pollution (such as aerosol pollution) caused by opening the cap again and problems such as false positive caused thereby.

Claims

1. A component for a reaction tube cap, characterized by, The reaction tube cap component comprises a body in the shape of a hollow cone or a hollow arc, comprising an upper end opening and a lower end aperture, the size of the body being suitable for being clamped in the cap cavity of the reaction tube cap, the lower end aperture having a hole diameter of 1-3 mm.

2. The component for a reaction tube cap according to claim 1, wherein The upper end outer diameter of the body is 4-6 mm.

3. The component for a reaction tube cap according to claim 1, wherein The upper end outer diameter of the body matches the inner diameter of the cap cavity of the reaction tube cap.

4. The component for a reaction tube cap according to claim 1, wherein The capacity of the reaction tube cap component is 0.5-20 μL.

5. The component for a reaction tube cap according to claim 1, wherein The reaction tube cap component is made of polypropylene.

6. The component for a reaction tube cap according to claim 1, wherein The reaction tube cap component is white or transparent.

7. The component for a reaction tube cap according to claim 1, wherein The reaction tube cap component further comprises a through-hole tube body, the upper end of the through-hole tube body being in an integral structure with the lower end of the body, the through-hole tube body having a through-hole penetrating through the upper end and the lower end, the through-hole tube body being in communication with the lower end aperture of the body.

8. The component for a reaction tube cap according to claim 7, wherein The through-hole tube body is 5-10 mm long, the inner diameter of the through-hole tube body being the same as the hole diameter of the lower end aperture.

9. A reaction tube cap comprising a reaction tube cap body, the reaction tube cap body comprising a cap cavity, characterized by, The reaction tube cap component can be clamped in the cap cavity, or the reaction tube cap component is arranged in the cap cavity and in an integral structure, the reaction tube cap component being as claimed in any one of claims 1-8.

10. A reaction tube comprising a reaction tube cap and a reaction tube body, the reaction tube cap being adapted to the reaction tube body, characterized in that, The reaction tube cap is as claimed in claim 9.

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