CN219637217U - Reaction tube for detecting loop-mediated isothermal amplification product - Google Patents
Reaction tube for detecting loop-mediated isothermal amplification product Download PDFInfo
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- CN219637217U CN219637217U CN202320172727.2U CN202320172727U CN219637217U CN 219637217 U CN219637217 U CN 219637217U CN 202320172727 U CN202320172727 U CN 202320172727U CN 219637217 U CN219637217 U CN 219637217U
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- China
- Prior art keywords
- reaction
- tube
- isothermal amplification
- mediated isothermal
- liquid storage
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- 238000006243 chemical reaction Methods 0.000 title claims abstract description 85
- 238000007397 LAMP assay Methods 0.000 title claims abstract description 43
- 239000007788 liquid Substances 0.000 claims abstract description 36
- 239000012295 chemical reaction liquid Substances 0.000 claims abstract description 21
- 238000001514 detection method Methods 0.000 claims abstract description 18
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 238000007789 sealing Methods 0.000 claims description 12
- 239000012188 paraffin wax Substances 0.000 claims description 8
- 239000002480 mineral oil Substances 0.000 claims description 5
- 235000010446 mineral oil Nutrition 0.000 claims description 5
- CGNLCCVKSWNSDG-UHFFFAOYSA-N SYBR Green I Chemical group CN(C)CCCN(CCC)C1=CC(C=C2N(C3=CC=CC=C3S2)C)=C2C=CC=CC2=[N+]1C1=CC=CC=C1 CGNLCCVKSWNSDG-UHFFFAOYSA-N 0.000 claims description 4
- 238000005192 partition Methods 0.000 claims 4
- 238000002844 melting Methods 0.000 claims 1
- 230000003321 amplification Effects 0.000 abstract description 15
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 10
- 238000001917 fluorescence detection Methods 0.000 abstract description 7
- 239000007850 fluorescent dye Substances 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 3
- 230000000007 visual effect Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 15
- 239000000443 aerosol Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 239000000243 solution Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The utility model discloses a reaction tube for detecting loop-mediated isothermal amplification products, which comprises a tube body and a tube cover, wherein a closed reaction cavity is formed between the tube body and the tube cover, a liquid storage cavity connected with the inner wall of the tube body is arranged at the upper part of the reaction cavity, the liquid storage cavity is arranged in an upward opening way, the open end of the liquid storage cavity is communicated with the reaction cavity, the reaction cavity is used for placing loop-mediated isothermal amplification reaction liquid therein, and the liquid storage cavity is used for placing detected color reagent therein. According to the utility model, through the design of the liquid storage cavity, the separation of the reaction system and the fluorescence detection system is realized, and after the reaction is finished, the combination of the amplification product and the fluorescent dye can be realized by reversing and uniformly mixing, so that the LAMP uncapped visual detection is realized.
Description
Technical Field
The utility model belongs to the technical field of loop-mediated isothermal amplification, and particularly relates to a reaction tube for detecting loop-mediated isothermal amplification products.
Background
Nucleic acid amplification is one of the most commonly used methods in gene diagnosis technology, and at present, the methods for nucleic acid amplification include Polymerase Chain Reaction (PCR), nucleic acid sequence-dependent amplification (NASBA), autonomous Sequence Replication (SR), strand displacement technology (SDA), etc., which can rapidly amplify a trace sample, but have various disadvantages in terms of specificity, simplicity, temperature, reagent instrument requirements, etc. Notomi et al in 2000 reported a novel nucleic acid amplification technique, namely, loop-mediated isothermal amplification (loop mediated isothermal amplification, LAMP) technique. The LAMP technology is a novel nucleic acid amplification technology, has the advantages of simplicity, rapidness, strong specificity, high sensitivity and the like, and has wide application prospect.
At present, detection modes of LAMP amplification products mainly comprise electrophoresis detection, fluorescence detection and ultraviolet absorption detection. The electrophoresis detection needs to take out and detect the reaction product after the reaction is finished, so that aerosol pollution is easy to generate. Most of fluorescent detection does not need to be carried out by uncovering at present, but fluorescent dye is added into a reaction system in advance to inhibit amplification reaction, so that amplification efficiency is reduced, and detection sensitivity is affected; if the fluorescent color-developing agent is added after the reaction is finished, the problem of aerosol pollution can also occur. The turbidimeter based on ultraviolet absorption detection design can realize uncapped detection of amplified products, but needs professional turbidimeter equipment, increases detection cost, and is inconvenient for performing on-site inspection. Therefore, how to solve the LAMP aerosol pollution and realize the uncapped detection of LAMP amplification products is a technical problem that needs to be solved by the skilled person.
Disclosure of Invention
The utility model aims to provide a reaction tube for detecting loop-mediated isothermal amplification products, which solves the problem of aerosol pollution in the prior art when the loop-mediated isothermal amplification products are detected. In order to achieve the above purpose, the specific technical scheme of the utility model is as follows:
the reaction tube for detecting the loop-mediated isothermal amplification product comprises a tube body and a tube cover, wherein a closed reaction cavity is formed between the tube body and the tube cover, a liquid storage cavity connected with the inner wall of the tube body is arranged at the upper part of the reaction cavity, the liquid storage cavity is arranged upwards in an opening manner, the opening end of the liquid storage cavity is communicated with the reaction cavity, loop-mediated isothermal amplification reaction liquid is placed in the reaction cavity, and a detected color reagent is placed in the liquid storage cavity.
Further, the liquid storage cavity is arranged upwards in an opening mode, and the liquid storage cavity and the pipe body are of an integrated structure.
Further, the upper part of the inner wall of the tube body is provided with an upward inclined baffle plate, the bottom of the baffle plate is fixedly connected with the inner wall of the tube body, the top of the baffle plate is accommodated in the reaction cavity, and an open liquid storage cavity is formed between the baffle plate and the inner wall of the tube body.
Further, a groove is formed in the upper portion of the inner wall of the tube body, one side wall of the groove is fixedly connected with the inner wall of the tube body, the other side wall of the groove is accommodated in the reaction cavity, and a liquid storage cavity for accommodating a color developing agent is formed in the groove.
Further, a sealing layer is arranged on the loop-mediated isothermal amplification reaction liquid.
Further, the sealing layer is mineral oil or low-melting-point paraffin.
Further, the color reagent is SYBR Green I.
Further, the tube cover is provided in an upwardly convex arc shape.
Compared with the prior art, the utility model has the following advantages:
(1) Through the design of the liquid storage cavity, the separation of a reaction system and a fluorescence detection system is realized, and after the reaction is finished, the combination of an amplification product and a fluorescent dye can be realized by reversing and uniformly mixing, so that the LAMP uncapping visual detection is realized;
(2) The sealing layer (such as paraffin and the like) is arranged in the reaction tube, so that the paraffin floats above the reaction system in the reaction process due to the small density, and the separation of the reaction system (loop-mediated isothermal amplification reaction liquid) and the fluorescence detection system (color reagent) is effectively realized; meanwhile, after the color development reaction is finished, the paraffin can be condensed again after the reaction tube is cooled, so that the liquid level is isolated from the air, and the possible hysteresis pollution caused by reaction products is effectively avoided.
(3) The reaction tube can carry out the LAMP amplification reaction and the fluorescence detection reaction in the same tube, can detect the LAMP amplification product without opening a reaction tube cover, effectively avoids the problem of aerosol pollution while saving operation steps, and simultaneously avoids the inhibition of the amplification reaction caused by the fact that fluorescent dye is added into a reaction system in advance; and the LAMP technology established by the reaction tube is simple and convenient to operate, low in detection cost, short in detection time, free of special instruments and suitable for on-site detection.
Drawings
FIG. 1 is a schematic diagram of the hollow tube structure of the reaction tube of the present utility model;
FIG. 2 is a schematic representation of the use of a reactor tube according to the present utility model;
FIG. 3 is a schematic view of another hollow tube structure of the reaction tube of the present utility model;
FIG. 4 is a schematic view showing the process of using the reaction tube of the present utility model.
In the figure: 1-tube body, 2-tube cover, 3-baffle, 4-recess, 10-reaction chamber, 30-stock solution chamber.
Description of the embodiments
The reaction tube for detecting loop-mediated isothermal amplification products according to the present utility model will be further described with reference to the accompanying drawings.
Examples
The reaction tube for detecting loop-mediated isothermal amplification products shown in fig. 1-2 comprises a tube body 1 and a tube cover 2, wherein a closed reaction cavity 10 is formed between the tube body 1 and the tube cover 2, a liquid storage cavity 30 connected with the inner wall of the tube body 1 is arranged at the upper part of the reaction cavity 10, the liquid storage cavity 30 is arranged in an upward opening manner, the open end of the liquid storage cavity 30 is communicated with the reaction cavity 10, loop-mediated isothermal amplification reaction liquid is placed in the reaction cavity 10, and a detected color reagent is placed in the liquid storage cavity 30.
Specifically, the upper portion of the inner wall of the tube body 1 is provided with an upward inclined baffle plate 3, a reaction cavity 10 is arranged in the tube body 1, and an open liquid storage cavity 30 is formed between the baffle plate 3 and the inner wall of the tube body 1. The baffle 3 and the pipe body 1 are integrally formed, the bottom of the baffle 3 is fixedly connected with the inner wall of the pipe body 1, and the top of the baffle 3 is accommodated in the reaction cavity 10. The bottom of the reaction chamber 10 is used for placing a loop-mediated isothermal amplification reaction liquid, and a sealing layer is arranged on the reaction liquid so as to seal the loop-mediated isothermal amplification reaction liquid in the sealing layer, wherein the sealing layer is preferably mineral oil or low-melting-point paraffin. The reservoir 30 is configured to hold a detection reagent, preferably SYBR Green I.
In order to facilitate the fusion of the color-developing agent and the loop-mediated isothermal amplification reaction solution, the tube cover 2 is preferably provided in an upwardly convex arc shape.
The specific use of the reaction tube for detecting loop-mediated isothermal amplification products as shown in FIG. 4 comprises the following steps:
(1) Opening a tube cover 2 of the reaction tube, adding a loop-mediated isothermal amplification reaction liquid into the reaction cavity 10, adding a drop of mineral oil or low-melting-point paraffin above the loop-mediated isothermal amplification reaction liquid, adding a proper amount of color reagent into the liquid storage cavity 30, and closing the tube cover 2;
(2) After sample addition is finished, the reaction tube is placed in a water bath to start reaction, and in the reaction process, a sealing layer is covered above the loop-mediated isothermal amplification reaction liquid because the loop-mediated isothermal amplification reaction liquid and the color developing agent are positioned in two different cavities, so that inhibition of amplification caused by the color developing agent fused into the loop-mediated isothermal amplification reaction liquid is effectively avoided;
(3) After the reaction is finished, the reaction tube is reversely vibrated, then the reaction tube is positively vibrated, so that the amplified product and the color reagent are fully mixed, the mixture is placed at room temperature, and the color development result is observed.
Examples
The reaction tube for detecting loop-mediated isothermal amplification products shown in fig. 3 comprises a tube body 1 and a tube cover 2, wherein a closed reaction cavity 10 is formed between the tube body 1 and the tube cover 2, a liquid storage cavity 30 connected with the inner wall of the tube body 1 is arranged at the upper part of the reaction cavity 10, the liquid storage cavity 30 is arranged in an upward opening manner, the open end of the liquid storage cavity 30 is communicated with the reaction cavity 10, loop-mediated isothermal amplification reaction liquid is placed in the reaction cavity 10, and a detected color reagent is placed in the liquid storage cavity 30.
The liquid storage cavity 30 is arranged upwards in an opening way, the liquid storage cavity 30 and the pipe body 1 are of an integrated structure, and the color developing agent in the liquid storage cavity 30 is preferably SYBR Green I. Specifically, the upper portion of the inner wall of the tube body 1 is provided with a groove 4, one side wall of the groove 4 is fixedly connected with the inner wall of the tube body 1, the other side wall of the groove is accommodated in a reaction cavity 10, a liquid storage cavity 30 for accommodating a color developing agent is arranged in the groove 4, the bottom of the reaction cavity 10 is used for accommodating loop-mediated isothermal amplification reaction liquid, a sealing layer is arranged on the reaction liquid to seal the loop-mediated isothermal amplification reaction liquid in the sealing layer, and mineral oil or low-melting-point paraffin is preferably used as the sealing layer.
In this embodiment, the groove 4 may be an arc groove connected to the inner wall of a part of the pipe body 1, or may be an annular groove connected to the inner wall of the whole pipe body. In order to facilitate the fusion of the color-developing agent and the loop-mediated isothermal amplification reaction solution, the tube cover 2 is preferably provided in an upwardly convex arc shape.
In summary, the liquid storage cavity 30 may have other structures, and only needs to separate the reaction system (loop-mediated isothermal amplification reaction liquid) from the fluorescence detection system (color-developing agent) during the sample addition reaction, and when the detection result is observed, the reaction system (loop-mediated isothermal amplification reaction liquid) and the fluorescence detection system (color-developing agent) can be fused by inversion.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present utility model, and not for limiting the same; although the utility model has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the utility model.
Claims (8)
1. The utility model provides a detect reaction tube of loop-mediated isothermal amplification product, includes body (1) and tube cap (2), its characterized in that forms inclosed reaction chamber (10) between body (1) and tube cap (2), reaction chamber (10) upper portion set up liquid storage chamber (30) with body (1) inner wall connection, liquid storage chamber (30) are upwards open setting, and open end communicates with each other with reaction chamber (10), be used for placing loop-mediated isothermal amplification reaction liquid in reaction chamber (10), be used for placing the developer of detection in liquid storage chamber (30).
2. The reaction tube for detecting loop-mediated isothermal amplification products according to claim 1, wherein the liquid storage cavity (30) and the tube body (1) are of an integrally formed structure.
3. The reaction tube for detecting loop-mediated isothermal amplification products according to claim 2, wherein an upward inclined partition plate (3) is arranged on the upper portion of the inner wall of the tube body (1), the bottom of the partition plate (3) is fixedly connected with the inner wall of the tube body (1), the top of the partition plate (3) is accommodated in the reaction cavity (10), and an open liquid storage cavity (30) is formed between the partition plate (3) and the inner wall of the tube body (1).
4. The reaction tube for detecting loop-mediated isothermal amplification products according to claim 2, wherein a groove (4) is formed in the upper portion of the inner wall of the tube body (1), one side wall of the groove (4) is fixedly connected with the inner wall of the tube body (1), the other side wall is accommodated in the reaction cavity (10), and a liquid storage cavity (30) for accommodating a color developing agent is formed in the groove (4).
5. The reaction tube for detecting loop-mediated isothermal amplification products according to claim 1, wherein a sealing layer is arranged on the loop-mediated isothermal amplification reaction liquid in the reaction cavity (10).
6. The reaction tube for detecting loop-mediated isothermal amplification products according to claim 5, wherein the sealing layer is mineral oil or low-melting paraffin.
7. The reaction tube for detecting loop-mediated isothermal amplification products according to any one of claims 1 to 6, wherein the chromogenic agent is SYBR Green I.
8. A reaction tube for detecting loop-mediated isothermal amplification products according to any of claims 1-6, wherein the tube cover (2) is provided in the shape of an upwardly convex arc.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320172727.2U CN219637217U (en) | 2023-02-10 | 2023-02-10 | Reaction tube for detecting loop-mediated isothermal amplification product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320172727.2U CN219637217U (en) | 2023-02-10 | 2023-02-10 | Reaction tube for detecting loop-mediated isothermal amplification product |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219637217U true CN219637217U (en) | 2023-09-05 |
Family
ID=87818508
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320172727.2U Active CN219637217U (en) | 2023-02-10 | 2023-02-10 | Reaction tube for detecting loop-mediated isothermal amplification product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN219637217U (en) |
-
2023
- 2023-02-10 CN CN202320172727.2U patent/CN219637217U/en active Active
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