Detection tube
By designing a detection tube with a sealed connection between the inner and outer tubes, operation without opening the cap is achieved, solving the contamination problem in CRISPR nucleic acid detection and improving detection efficiency and accuracy.
Patent Information
- Application Number
- CN202422333386.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-25
- Publication Date
- 2025-11-07
- Estimated Expiration
- 2034-09-25
AI Technical Summary
Existing CRISPR nucleic acid detection technology requires opening and closing the cap after nucleic acid amplification, which can easily lead to aerosol contamination and cross-contamination in the laboratory, affecting the accuracy and efficiency of the test results.
A detection tube was designed with a sealed connection between the inner and outer tubes. The inner tube contains nucleic acid amplification reagents and sample nucleic acid, while the outer tube contains CRISPR reagents. The tube cap can squeeze the inner tube to break it, transferring the sample solution to the outer tube, thus achieving operation without opening the cap and reducing the risk of contamination.
Without reducing detection sensitivity, the experimental procedures were reduced, contamination was prevented, and the efficiency and accuracy of nucleic acid testing were improved.
Smart Images
Figure CN223522536U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to nucleic acid molecule diagnosis experimental equipment technical field, especially a detection tube. BACKGROUND
[0002] At present, in the field of human infectious disease diagnosis, food safety, pathogen determination, global biological safety and environmental analysis tracking biological pollution and environmental quality monitoring, rapid, accurate, sensitive and quantitative detection of specific nucleic acid sequences is indispensable.
[0003] CRISPR is the full name of Clustered regularly interspaced short palindromic repeats, and the full name in Chinese is regular clustered interspaced short palindromic repeat. It is a kind of adaptive immune system widely existing in ancient bacteria and bacteria. In the prior art, after the CRISPR technology is applied to the field of nucleic acid detection, various nucleic acid detection platforms based on Cas12, Cas13, Cas14 and other CRISPR enzymes also emerge as the times require. CRISPR technology shows strong advantages in detection cost, efficiency, portability, specificity and simplicity, and has stronger scene application. At the same time, it can be combined with other technologies, so as to more simply and highly sensitively detect nucleic acid.
[0004] However, there are still some problems in the application of CRISPR technology, such as: before the fluorescence value of CRISPR nucleic acid detection, the nucleic acid amplification product needs to be added into the CRISPR reagent to realize the detection of the target. The opening and closing of the cover after nucleic acid amplification operation is easy to cause laboratory aerosol pollution, which not only leads to higher false positive results in subsequent results, but also causes cross contamination of laboratory and related equipment consumables. Utility model content
[0005] The utility model aims at providing a detection tube to solve the problems existing in the prior art, which can reduce experimental operation, prevent pollution and improve the efficiency and accuracy of nucleic acid detection.
[0006] To achieve the above-mentioned purpose, the utility model provides the following scheme:
[0007] The utility model provides a kind of detection tube, including outer tube, inner tube and pipe cover, the top end opening of outer tube, the outer tube can accommodate CRISPR reagent, the inner tube can accommodate nucleic acid amplification reagent and the sample nucleic acid to be detected, the inner tube is used to be fixed in the outer tube, and the inner tube is placed above the CRISPR reagent contained in the outer tube, the outer wall of the inner tube is sealed with the inner wall of the outer tube, the pipe cover is placed above the inner tube, the bottom end of the pipe cover is used to be inserted into the outer tube from the top end opening of the outer tube, the outer wall of the part on the pipe cover placed in the outer tube is sealed with the inner wall of the outer tube, the bottom end of the pipe cover can extrude the inner tube and make the inner tube break, so that sample liquid in the inner tube falls into the CRISPR reagent contained in the outer tube.
[0008] Preferably, the inner wall of the outer tube has a first limiting step, the space below the first limiting step in the outer tube is a first containing space, the first containing space is used to contain the CRISPR reagent, the first limiting step is connected with the inner tube, and the first limiting step can bear the inner tube.
[0009] Preferably, the inner tube includes a top column and a sleeve, the bottom surface of the sleeve is in contact with the top surface of the first limiting step, the sleeve is sleeved outside the top column, the bottom of the sleeve is sealingly and fixedly connected with the bottom of the top column, and the bottom of the sleeve has a weak connection part connected with the bottom of the top column.
[0010] Preferably, the thickness of the weak connection part is 0.05-0.07mm.
[0011] Preferably, the inner tube further includes a hollow anti-falling frame, the top end of the hollow anti-falling frame is fixedly connected with the bottom end of the sleeve, and the hollow anti-falling frame can receive the top column.
[0012] Preferably, the top column is a first material top column, the sleeve is a second material sleeve, and the first material is different from the second material.
[0013] Preferably, the volume of the outer tube is 200μL, the top column is a hexagonal prism, the length of the top column is 5-10mm, and the length of the sleeve is 1-5mm.
[0014] Preferably, a positioning groove is formed in the pipe cover, the positioning groove can be in contact with the top end of the top column, and the top end of the top column is inserted into the positioning groove.
[0015] Preferably, the inner wall of the outer tube has a second limiting step, the bottom end of the pipe cover has an annular limiting protrusion, and the annular limiting protrusion can be in contact with the top surface of the second limiting step or the top surface of the sleeve.
[0016] Preferably, it also includes a connecting strap, one end of which is fixedly connected to the top end of the outer tube, and the other end of which is fixedly connected to the tube cap.
[0017] The present invention achieves the following technical advantages over the prior art:
[0018] The detection tube provided by this utility model has an outer tube containing CRISPR reagents and an inner tube containing nucleic acid amplification reagents and the nucleic acid sample to be tested. The inner tube serves both to store the nucleic acid amplification reagents used in the nucleic acid amplification reaction and as a reaction container for the nucleic acid amplification reaction. The outer wall of the inner tube is sealed to the inner wall of the outer tube to prevent the CRISPR reagents in the outer tube from escaping into the inner tube and causing interference. The outer wall of the portion of the tube cap placed inside the outer tube is sealed to the inner wall of the outer tube, ensuring the airtightness between the tube cap, the inner tube, and the outer tube after the tube cap is closed. The bottom end of the tube cap can squeeze the inner tube and cause it to break, allowing the sample liquid in the inner tube to fall into the CRISPR reagent contained in the outer tube. The detection can be completed without opening the cap. The detection tube provided by this utility model can ensure that nucleic acid detection based on CRISPR technology can reduce experimental operations, prevent contamination, and improve the efficiency and accuracy of nucleic acid detection without reducing sensitivity or causing a high false positive rate. Attached Figure Description
[0019] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0020] Figure 1 A schematic diagram of the structure of the detection tube provided by this utility model;
[0021] Figure 2 for Figure 1 Schematic diagram of the inner tube;
[0022] Figure 3 for Figure 1 Schematic diagram of the outer pipe and pipe cap;
[0023] Figure 4 for Figure 1 A top view of the detection tube in the diagram;
[0024] Figure 5 for Figure 1 A schematic diagram of the first stage of the tube cap of the detection tube in the middle;
[0025] Figure 6 for Figure 1 A schematic diagram of the second stage of the tube cap of the detection tube in the middle;
[0026] In the diagram: 1-outer tube, 2-inner tube, 3-tube cap, 4-nucleic acid amplification reagent and nucleic acid of the sample to be tested, 5-CRISPR reagent, 6-first limiting step, 7-first accommodating space, 8-top column, 9-sleeve, 10-weak connection, 11-hollowed-out anti-fall frame, 12-connecting strap. Detailed Implementation
[0027] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.
[0028] The purpose of this invention is to provide a detection tube that solves the problems existing in the prior art, reduces experimental operations, prevents contamination, and improves the efficiency and accuracy of nucleic acid detection.
[0029] To make the above-mentioned objectives, features and advantages of this utility model more apparent and understandable, the utility model will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0030] like Figures 1-6 As shown, this utility model provides a detection tube, including an outer tube 1, an inner tube 2, and a cap 3. The top end of the outer tube 1 is open, and the outer tube 1 can contain CRISPR reagent 5. The inner tube 2 can contain nucleic acid amplification reagent and the nucleic acid of the sample to be tested 4. The inner tube 2 is fixed inside the outer tube 1 and is positioned above the CRISPR reagent 5 contained in the outer tube 1. The outer wall of the inner tube 2 is sealed and fitted to the inner wall of the outer tube 1. The cap 3 is positioned above the inner tube 2. The bottom end of the cap 3 is used to extend into the outer tube 1 from the top end opening of the outer tube 1. The outer wall of the portion of the cap 3 placed inside the outer tube 1 is sealed and connected to the inner wall of the outer tube 1. The bottom end of the cap 3 can squeeze the inner tube 2 and cause the inner tube 2 to break, so that the sample liquid in the inner tube 2 falls into the CRISPR reagent 5 contained in the outer tube 1.
[0031] The detection tube provided by the utility model, the outer tube 1 contains CRISPR reagent 5, the inner tube 2 contains nucleic acid amplification reagent and sample nucleic acid 4 to be detected, the inner tube 2 is used to store nucleic acid amplification reagent for nucleic acid amplification reaction and is used as the reaction container of nucleic acid amplification reaction, the outer wall of the inner tube 2 is sealed and matched with the inner wall of the outer tube 1, the interference caused by the escape of CRISPR reagent 5 in the outer tube 1 to the inner tube 2 is avoided, the outer wall of the part of the tube cover 3 placed in the outer tube 1 is sealed and connected with the inner wall of the outer tube 1, the sealing property between the tube cover 3 and the inner tube 2 and the outer tube 1 after covering the tube cover 3 can be guaranteed, the bottom end of the tube cover 3 can extrude the inner tube 2 and make the inner tube 2 break, so that the sample liquid in the inner tube 2 falls into the CRISPR reagent 5 contained in the outer tube 1, the detection can be completed without opening the cover, the detection tube provided by the utility model can guarantee that the nucleic acid detection based on CRISPR technology does not reduce the sensitivity and does not cause higher false positives, reduces experimental operation, prevents pollution, improves the efficiency and accuracy of nucleic acid detection.
[0032] As a more preferred embodiment of the present embodiment, the outer tube 1 is a conical tube with a closed bottom, which is simple in structure and convenient to manufacture and use.
[0033] As a more preferred embodiment of the present embodiment, the inner wall of the outer tube 1 has a first limiting step 6, the space below the first limiting step 6 in the outer tube 1 is a first containing space 7, the first containing space 7 is used to contain CRISPR reagent 5, the volume of the first containing space 7 is preferably 10-100 muL, more preferably 25 muL, the first limiting step 6 is connected with the inner tube 2, and the first limiting step 6 can bear the inner tube 2, which is simple in structure and convenient to manufacture and use.
[0034] As a more preferred embodiment of the present embodiment, the inner tube 2 includes a top column 8 and a sleeve 9, the bottom surface of the sleeve 9 is in contact with the top surface of the first limiting step 6, which is simple in structure and convenient to use, the sleeve 9 is sleeved on the outside of the top column 8, the bottom of the sleeve 9 is sealingly and fixedly connected with the bottom of the top column 8, the space surrounded by the sleeve 9 and the top column 8 is a storage space for nucleic acid amplification reagent and sample nucleic acid 4 to be detected, and the volume is preferably 10-100 muL, more preferably 40 muL, and the bottom of the sleeve 9 and the bottom of the top column 8 have a weak connection part 10, which is more easily pressed and broken.
[0035] As a more preferred embodiment of the present embodiment, the thickness of the weak connection part 10 is 0.05-0.07 mm, and the weak connection part 10 is a circular ring with a thickness of 0.06 mm.
[0036] As a more preferred embodiment of the present embodiment, the inner tube 2 further comprises a hollow anti-falling frame 11, the top end of the hollow anti-falling frame 11 is fixedly connected with the bottom end of the sleeve 9, and the hollow anti-falling frame 11 can accommodate the top column 8, so as to avoid the top column 8 from falling to the CRISPR reagent 5, thereby avoiding affecting the subsequent detection.
[0037] As a more preferred embodiment of the present embodiment, the top column 8 is a first material top column, the sleeve 9 is a second material sleeve, the first material is different from the second material, and according to different characteristics of different materials, the weak connection part 10 at the connection between the bottom of the sleeve 9 and the bottom of the top column 8 can be easily separated by pressing the top column 8 through the tube cover 3, and the top column 8 and the sleeve 9 are prevented from being adhered after being separated.
[0038] As a more preferred embodiment of the present embodiment, the volume of the outer tube 1 is 200 μL, the top column 8 is a hexagonal prism, the length of the top column 8 is 5-10 mm, preferably 9 mm, and the length of the sleeve 9 is 1-5 mm, preferably 3 mm.
[0039] As a more preferred embodiment of the present embodiment, the tube cover 3 is provided with a positioning groove, the top end of the top column 8 can be in contact with the positioning groove, and the top end of the top column 8 can be inserted into the positioning groove, thereby improving the contact reliability of the tube cover 3 and the top column 8.
[0040] As a more preferred embodiment of the present embodiment, the inner wall of the outer tube 1 is provided with a second limiting step, the bottom end of the tube cover 3 is provided with an annular limiting protrusion, the annular limiting protrusion can be in contact with the top surface of the second limiting step or the top surface of the sleeve 9, the movement of the tube cover 3 can be effectively limited, and the top column 8 can be prevented from falling to the CRISPR reagent 5, thereby avoiding affecting the subsequent detection.
[0041] Further, the detection tube provided by the utility model further comprises a connecting belt 12, one end of the connecting belt 12 is fixedly connected with the top end of the outer tube 1, the other end of the connecting belt 12 is fixedly connected with the tube cover 3, as a more preferred embodiment of the present embodiment, the outer tube 1, the connecting belt 12 and the tube cover 3 are all made of polypropylene material and are integrally formed, the connecting belt 12 is 0.1-1 cm longer than the connecting strip of a conventional eight connecting pipe, and is more preferably 0.5 cm longer.
[0042] The pressing of the tube cover 3 has two stages, the first stage is that the tube cover 3 contacts the top column 8, and the second stage is that the top column 8 presses and breaks the weak connection part 10, when the tube cover 3 is pressed to the first stage and the second stage, the outer wall of the part of the tube cover 3 in the outer tube 1 is in sealing connection with the inner wall of the outer tube 1.
[0043] The method for performing CRISPR molecular diagnosis by using the detection tube comprises the following steps: adding CRISPR reagent 5 (such as 36 μL) into the outer tube 1, then fixing the inner tube 2 in the outer tube 1, and adding nucleic acid amplification reagent (such as 13 μL) into the inner tube 2; adding sample nucleic acid (such as 2 μL) to be detected into the inner tube 2, covering the tube cover 3, and pressing the tube cover 3 to the first stage; performing nucleic acid constant temperature amplification program, the amplification temperature is 36-42 DEG C, the duration is 10-30 min, preferably 37 DEG C, 12 min; after the nucleic acid amplification is completed, the tube cover 3 is pressed once, the tube cover 3 is pressed to the second stage, the weak connection part 10 is broken, the amplification product falls to the CRISPR reagent 5, then the inner tube 2 is shaken manually for 3-10 times, or instantaneous centrifugation is performed, the rotation speed is 100-3000 rpm, the time is 5-20 s, preferably 2000 rpm, 10 s, the CRISPR nucleic acid detection program is performed, the detection temperature is 36-42 DEG C, the duration is 15-50 min, preferably 37 DEG C, 25 min, and then the fluorescence value is detected.
[0044] The principle and implementation mode of the specific examples are described in the utility model, and the above embodiment is only used for helping to understand the method and core idea of the utility model; meanwhile, according to the idea of the utility model, the specific implementation mode and application range will be changed by the general technical personnel in the field; according to the above, the content of the specification should not be understood as the limitation of the utility model.
Claims
1. A detection tube, characterized by: The application relates to a CRISPR reagent container, which comprises an outer tube, an inner tube and a tube cover, wherein the top end of the outer tube is open, the outer tube can contain CRISPR reagents, the inner tube can contain nucleic acid amplification reagents and sample nucleic acids to be detected, the inner tube is arranged in the outer tube, and the inner tube is arranged above the CRISPR reagents contained in the outer tube; the outer wall of the inner tube is tightly combined with the inner wall of the outer tube; the tube cover is arranged above the inner tube, the bottom end of the tube cover is arranged in the outer tube from the top end opening of the outer tube, the outer wall of the part of the tube cover arranged in the outer tube is tightly connected with the inner wall of the outer tube, and the bottom end of the tube cover can extrude the inner tube and make the inner tube break, so that the sample liquid in the inner tube falls into the CRISPR reagents contained in the outer tube; the inner tube comprises a top column and a sleeve, the sleeve is arranged outside the top column, the bottom of the sleeve is tightly fixedly connected with the bottom of the top column, and the bottom of the sleeve is connected with the bottom of the top column and has a weak connection part; the inner tube further comprises a hollow anti-falling frame, the top end of the hollow anti-falling frame is fixedly connected with the bottom end of the sleeve, and the hollow anti-falling frame can support the top column; a positioning groove is arranged on the tube cover, the top end of the top column can be in contact with the positioning groove, and the top end of the top column is arranged in the positioning groove.
2. The test tube of claim 1, wherein: The inner wall of the outer tube is provided with a first limiting step, the space below the first limiting step in the outer tube is a first containing space, the first containing space is used for containing the CRISPR reagents, the first limiting step is connected with the inner tube, and the first limiting step can support the inner tube.
3. The test tube of claim 2, wherein: The bottom surface of the sleeve is in contact with the top surface of the first limiting step.
4. The test tube of claim 3, wherein: The thickness of the weak connection part is 0.05-0.07 mm.
5. The test tube of claim 3, wherein: The top column is a top column made of a first material, and the sleeve is a sleeve made of a second material; the first material is different from the second material.
6. The test tube of claim 3, wherein: The volume of the outer tube is 200 muL, the top column is a hexagonal prism, the length of the top column is 5-10 mm, and the length of the sleeve is 1-5 mm.
7. The test tube of claim 3, wherein: The inner wall of the outer tube is provided with a second limiting step, the bottom end of the tube cover is provided with an annular limiting protrusion, and the annular limiting protrusion can be in contact with the top surface of the second limiting step or the top surface of the sleeve.
8. The test tube of claim 1, wherein: The application further comprises a connecting belt, one end of the connecting belt is fixedly connected with the top end of the outer tube, and the other end of the connecting belt is fixedly connected with the tube cover.