CN214300128U - Loop-mediated isothermal amplification reaction tube based on capillary tube - Google Patents
Loop-mediated isothermal amplification reaction tube based on capillary tube Download PDFInfo
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- CN214300128U CN214300128U CN202120066077.4U CN202120066077U CN214300128U CN 214300128 U CN214300128 U CN 214300128U CN 202120066077 U CN202120066077 U CN 202120066077U CN 214300128 U CN214300128 U CN 214300128U
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Abstract
The utility model discloses a loop-mediated isothermal amplification reaction tube based on capillary establishes the tube cap on the reaction body, and the reaction body is built-in to have loop-mediated isothermal amplification reaction system, and the bottom is provided with the rubber buffer in the tube cap, is provided with the capillary on the rubber buffer, and the last port of capillary and inside looks UNICOM of tube cap, lower port and reaction body looks UNICOM. The utility model discloses a loop-mediated isothermal amplification system and fluorochrome add reaction tube body and capillary respectively in, insert the capillary rubber buffer through in order being fixed, the fluorochrome that extrusion tube cap made in the capillary after the reaction finishes blows in the reaction tube body, mix with the reaction system liquid in the reaction tube body and take place the color-changing reaction, the realization is with the fluorochrome of appraising loop-mediated isothermal amplification reaction product in the capillary separately with the reaction system in the reaction tube body, add fluorochrome and lead to loop-mediated isothermal amplification reaction to be restrained before avoiding the reaction, thereby add fluorochrome because of uncapping and cause the pollution to make the reaction appear false positive result after the reaction.
Description
Technical Field
The utility model belongs to the technical field of reaction equipment or device, concretely relates to loop-mediated isothermal amplification reaction tube based on capillary.
Background
Loop-mediated isothermal amplification (LAMP) is a technique proposed by the japanese scholars in Notomi equal to 2000 that can rapidly amplify DNA fragments under constant temperature conditions, and the technique utilizes 4 specific primers and strand displacement DNA Polymerase (Bst DNA Polymerase) for 6 gene segments in a target sequence, and can complete amplification reaction under constant temperature conditions for tens of minutes even without the process of denaturation, annealing and cyclic temperature change of double-stranded template DNA through PCR reaction. In recent years, the technology is researched and applied to detection of bacteria, viruses, microorganisms and the like in a plurality of research fields such as food safety, medical inspection, planting, livestock raising, aquatic products and the like. LAMP research is widely applied to rapid detection of nucleic acid and is considered to have good application prospect. In the common detection methods of four amplification results of gel electrophoresis, fluorescent dye, biosensor method and turbidity method, the fluorescent dye does not need extra instruments, can be judged by observing the color change of the fluorescent dye, and is very convenient for field detection. SYBR Green I dye is used as a most commonly used color developing agent in LAMP product analysis, if the dye is added into an LAMP reaction system in advance, LAMP reaction can be severely inhibited, the color developing agent is added after the reaction, and high-concentration amplification products are easy to cause aerosol pollution when the tube opening operation is carried out, are difficult to remove and are easy to cause false positive detection results.
At present, in order to solve the problem of aerosol pollution caused by adding dye after reaction by opening a tube, related technical schemes are disclosed and used. In their patent, us issued 2014, a device comprising a reaction tube and a syringe is proposed, which is obtained by piercing a silicone plug provided at the nozzle of the reaction tube body with the syringe needle used. However, in the LAMP experiment, the usage amount of the fluorescent dye is very small, the sterile syringe is not easy to accurately absorb and add, and the aerosol can not be ensured to leak along with the syringe. In 2017, Xiaojiu et al proposed a reaction kit consisting of a common centrifugal outer tube and a paraffin inner tube in the patent, but the inner tube is made of a material with a melting point of not higher than 65 ℃, so that the inner tube is easy to solidify and is not easy to control in the production and manufacturing process, the temperature of LAMP reaction is also between 58 ℃ and 65 ℃, and the risk of leakage of fluorescent dye may exist. In 2018, a method for improving loop-mediated isothermal amplification false positive pollution by using isothermal amplification device equipment is proposed in a Liweihao patent, but the equipment used in the technology is complex, complete adsorption of aerosol by water vapor cannot be guaranteed in actual operation, and dropwise addition of a trace amount of liquid is not easy to control.
Disclosure of Invention
The utility model aims to solve the technical problem that overcome above-mentioned prior art equipment complicacy and be difficult for the shortcoming of control, nucleic acid amplification product hourglass outward, provide a simple structure, easy operation, small, accurate add fluorochrome, avoid aerosol pollution's loop-mediated isothermal amplification reaction tube based on capillary.
The technical scheme for solving the technical problems is as follows: the reaction tube body is provided with a tube cover, a loop-mediated isothermal amplification reaction system is arranged in the reaction tube body, the bottom in the tube cover is provided with a rubber plug, the rubber plug is provided with a capillary tube, the upper port of the capillary tube is communicated with the inside of the tube cover, and the lower port of the capillary tube is communicated with the reaction tube body.
The utility model discloses a tube cap is the rubber tube cap.
The capillary tube of the utility model is: be provided with the top tube on the low tube and top tube and low tube are even as an organic whole, and the internal diameter of top tube is 1.2 ~ 2mm, and the internal diameter of low tube is 0.5 ~ 0.9 mm.
The utility model discloses a be marked with scale mark 303 on the capillary outer wall.
The space between the plane of the bottom end face of the capillary and the plane of the upper end face of the loop-mediated isothermal amplification reaction system of the reaction tube body is not more than 1.5 cm.
The inner diameter of the reaction tube body of the utility model is 0.7-0.8 cm.
The reaction tube body of the utility model is connected with the tube cap through screw threads.
The utility model discloses a loop-mediated isothermal amplification system and fluorochrome add reaction tube body and capillary respectively in, insert the capillary rubber buffer through in order being fixed, the fluorochrome that extrusion tube cap made in the capillary after the reaction finishes blows in the reaction tube body, mix with the reaction system liquid in the reaction tube body and take place the color-changing reaction, can realize separately the fluorochrome of appraising loop-mediated isothermal amplification reaction product in the capillary with the reaction system in the reaction tube body, add fluorochrome and lead to loop-mediated isothermal amplification reaction to be restrained before avoiding the reaction, thereby add fluorochrome because of uncapping and cause the pollution to make the reaction appear false positive result after the reaction. The utility model has the advantages of simple structure, convenient operation and easy operation.
Drawings
Fig. 1 is a schematic structural diagram of an embodiment of the present invention.
Fig. 2 is a schematic view of the structure of the capillary tube 3 in fig. 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
As shown in FIGS. 1 and 2, the capillary-based loop-mediated isothermal amplification reaction tube of the present embodiment is composed of a tube cap 1, a rubber stopper 2, a capillary 3, and a reaction tube body 4 connected together.
The reaction tube body 4 is connected with a tube cap 1 through threads, and the tube cap 1 is a rubber tube cap. The inner diameter of the reaction tube body 4 is 0.75cm, a loop-mediated isothermal amplification reaction system is arranged in the reaction tube body 4, and the reaction carried out in the reaction tube body 4 is a loop-mediated isothermal nucleic acid amplification reaction. Bottom stopper has rubber buffer 2 in the tube cap 1, and fixed mounting has capillary 3 on the rubber buffer 2, and the material of capillary 3 is rigid plastic, can pierce through rubber buffer 2 and not broken, and the last port of capillary 3 and 1 inside looks UNICOM of tube cap, lower port and 4 looks UNICOMs of reaction body, capillary 3 are: an upper pipe 301 is arranged on a lower pipe 302, the upper pipe 301 is connected with the lower pipe 302 into a whole, the inner diameter of the upper pipe 301 is 1.5mm, the inner diameter of the lower pipe 302 is 0.7mm, the outer wall of a capillary 3 is marked with a scale mark 303, the scale mark 303 is used for accurately sucking and identifying fluorescent dye of a loop-mediated isothermal amplification reaction product, and the distance between the plane where the bottom end face of the capillary 3 is located and the plane where the upper end face of a loop-mediated isothermal amplification reaction system of a reaction pipe body 4 is located is not more than 1.5 cm. The utility model discloses can realize separately the fluorochrome of appraising among the capillary 3 loop-mediated isothermal amplification reaction product with the reaction system among the reaction body 4, add fluorochrome and lead to loop-mediated isothermal amplification reaction to be inhibited before avoiding reacting, blow out the fluorochrome in capillary 3 through the air in the extrusion tube cap 1, make its mix with the loop-mediated isothermal amplification reaction system among the reaction body 1, thereby avoided the reaction back because of uncapping adds fluorochrome and causes the pollution to make the reaction appear false positive result.
The operation steps of this embodiment are as follows:
before the loop-mediated isothermal nucleic acid amplification reaction, adding a reaction system of the loop-mediated isothermal amplification reaction into a reaction tube body 4, accurately absorbing a proper amount of fluorescent dye according to a scale mark 303 on a capillary tube 3, inserting the capillary tube 3 absorbing the fluorescent dye through a rubber plug 2 and fixing the capillary tube 3 on the rubber plug 2, hermetically connecting a tube cover 1 and the reaction tube body 4, after the reaction is finished, extruding the tube cover 1, applying air pressure to blow out the fluorescent dye in the capillary tube 3 into the reaction tube body 4, mixing the fluorescent dye with a reaction system liquid in the reaction tube body 4 to perform a color change reaction, and judging the nucleic acid amplification condition according to the color change.
Example 2
The reaction tube body 4 is connected with a tube cap 1 through threads, and the tube cap 1 is a rubber tube cap. The inner diameter of the reaction tube body 4 is 0.8cm, and a loop-mediated isothermal amplification reaction system is arranged in the reaction tube body 4. The bottom stopper has rubber buffer 2 in the tube cap 1, and fixed mounting has capillary 3 on the rubber buffer 2, and the last port of capillary 3 and 1 inside looks UNICOM of tube cap, lower port and 4 looks UNICOMs of reaction tube body, capillary 3 are: the upper tube 301 is arranged on the lower tube 302, the upper tube 301 and the lower tube 302 are connected into a whole, the inner diameter of the upper tube 301 is 1.2mm, the inner diameter of the lower tube 302 is 0.5mm, the outer wall of the capillary tube 3 is marked with a scale mark 303, and the distance between the plane where the bottom end face of the capillary tube 3 is located and the plane where the upper end face of the loop-mediated isothermal amplification reaction system of the reaction tube body 4 is located is not more than 1.5 cm.
Other components and their connection and operation steps are the same as those in embodiment 1.
Example 3
The reaction tube body 4 is connected with a tube cap 1 through threads, and the tube cap 1 is a rubber tube cap. The inner diameter of the reaction tube body 4 is 0.7cm, and a loop-mediated isothermal amplification reaction system is arranged in the reaction tube body 4. The bottom stopper has rubber buffer 2 in the tube cap 1, and fixed mounting has capillary 3 on the rubber buffer 2, and the last port of capillary 3 and 1 inside looks UNICOM of tube cap, lower port and 4 looks UNICOMs of reaction tube body, capillary 3 are: the upper tube 301 is arranged on the lower tube 302, the upper tube 301 and the lower tube 302 are connected into a whole, the inner diameter of the upper tube 301 is 2mm, the inner diameter of the lower tube 302 is 0.9mm, the outer wall of the capillary tube 3 is marked with a scale mark 303, and the distance between the plane where the bottom end face of the capillary tube 3 is located and the plane where the upper end face of the loop-mediated isothermal amplification reaction system of the reaction tube body 4 is located is not more than 1.5 cm.
Other components and their connection and operation steps are the same as those in embodiment 1.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention.
Claims (7)
1. A loop-mediated isothermal amplification reaction tube based on capillary tubes is characterized in that: the reaction tube body (4) is provided with a tube cover (1), a loop-mediated isothermal amplification reaction system is arranged in the reaction tube body (4), the bottom in the tube cover (1) is provided with a rubber plug (2), the rubber plug (2) is provided with a capillary tube (3), the upper port of the capillary tube (3) is communicated with the inside of the tube cover (1), and the lower port of the capillary tube (3) is communicated with the reaction tube body (4).
2. The capillary-based loop-mediated isothermal amplification reaction tube of claim 1, wherein: the pipe cover (1) is a rubber pipe cover.
3. The capillary-based loop-mediated isothermal amplification reaction tube according to claim 1, wherein the capillary (3) is: be provided with on lower tube (302) and go up pipe (301) and lower tube (302) even as an organic whole, the internal diameter of going up pipe (301) is 1.2 ~ 2mm, and the internal diameter of lower tube (302) is 0.5 ~ 0.9 mm.
4. The capillary-based loop-mediated isothermal amplification reaction tube according to claim 1 or 3, wherein: the outer wall of the capillary tube (3) is marked with a scale mark 303.
5. The capillary-based loop-mediated isothermal amplification reaction tube of claim 1, wherein: the distance between the plane of the bottom end face of the capillary tube (3) and the plane of the upper end face of the loop-mediated isothermal amplification reaction system of the reaction tube body (4) is not more than 1.5 cm.
6. The capillary-based loop-mediated isothermal amplification reaction tube of claim 1, wherein: the inner diameter of the reaction tube body (4) is 0.7-0.8 cm.
7. The capillary-based loop-mediated isothermal amplification reaction tube of claim 1, wherein: the reaction tube body (4) is connected with the tube cover (1) through threads.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202120066077.4U CN214300128U (en) | 2021-01-12 | 2021-01-12 | Loop-mediated isothermal amplification reaction tube based on capillary tube |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202120066077.4U CN214300128U (en) | 2021-01-12 | 2021-01-12 | Loop-mediated isothermal amplification reaction tube based on capillary tube |
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| Publication Number | Publication Date |
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| CN214300128U true CN214300128U (en) | 2021-09-28 |
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| CN202120066077.4U Active CN214300128U (en) | 2021-01-12 | 2021-01-12 | Loop-mediated isothermal amplification reaction tube based on capillary tube |
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| CN (1) | CN214300128U (en) |
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2021
- 2021-01-12 CN CN202120066077.4U patent/CN214300128U/en active Active
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