CN117165416A - Dilution tube, nucleic acid detection device and detection method thereof - Google Patents
Dilution tube, nucleic acid detection device and detection method thereof Download PDFInfo
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- CN117165416A CN117165416A CN202311453962.8A CN202311453962A CN117165416A CN 117165416 A CN117165416 A CN 117165416A CN 202311453962 A CN202311453962 A CN 202311453962A CN 117165416 A CN117165416 A CN 117165416A
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- 238000010790 dilution Methods 0.000 title claims abstract description 180
- 239000012895 dilution Substances 0.000 title claims abstract description 180
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 53
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 53
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 53
- 238000001514 detection method Methods 0.000 title claims abstract description 47
- 239000000243 solution Substances 0.000 claims abstract description 77
- 239000003085 diluting agent Substances 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000001125 extrusion Methods 0.000 claims abstract description 12
- 230000003321 amplification Effects 0.000 claims abstract description 8
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims description 58
- 238000012360 testing method Methods 0.000 claims description 19
- 238000011010 flushing procedure Methods 0.000 claims description 10
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000005070 sampling Methods 0.000 claims description 6
- 238000003780 insertion Methods 0.000 claims description 4
- 230000037431 insertion Effects 0.000 claims description 4
- 239000013013 elastic material Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 238000007789 sealing Methods 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 abstract description 2
- 238000007689 inspection Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract 1
- 238000013461 design Methods 0.000 description 10
- 239000000523 sample Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 2
- 241000700605 Viruses Species 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012938 design process Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000011900 installation process Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Classifications
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- 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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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The application discloses a dilution tube, a nucleic acid detection device and a detection method thereof; belongs to the field of medical examination and inspection instruments and molecular diagnosis and detection instruments; the technical key points are as follows: the dilution pipe includes: upper dilution pipe, lower dilution pipe, pipe plug, diluent. The solution in the dilution tube for pushing and sucking back the amplified product solution in the reaction tank for many times is 200-300 mu L by the diluent, and the solution in the dilution tube is discharged through x times of extrusion and loosening; the inner diameter of the extrusion part of the upper dilution tube is ymm, and the diameter of the inner diameter of the second space is zmm; the values of y and z need to satisfy the following formula: y= -0.25x 2 +3.25x‑0.5；z=0.005x 2 0.015x+0.55, and can sufficiently mix the amplification product solution uniformly. The present application aims to provide a dilution tube, a nucleic acid detecting apparatus and a method for detecting nucleic acidThe detection method can improve the accuracy of nucleic acid detection.
Description
Technical Field
The application relates to the field of medical examination and inspection instruments and molecular diagnosis and detection instruments, in particular to a dilution tube, a nucleic acid detection device and a detection method thereof.
Background
In the prior art, CN115125129B, a constant temperature chromatography nucleic acid detection device and a detection method thereof, CN115141745B, a multichannel split kit, a nucleic acid detection device and a detection method combine the ideas of fluorescence quantitative PCR and antigen detection, and a detection method capable of being applied to a household environment is provided. The method comprises the following steps of amplifying a sample by a heating component, diluting the concentration of the sample to a proper concentration by a diluent, then feeding a sample solution onto chromatographic test paper, and analyzing whether viruses exist in the solution by the chromatographic test paper.
The success or failure of the above scheme depends mainly on the following points:
first, the running speed of the solution on the chromatographic test paper;
second, whether the sample is leak-proof during the heat-up amplification process.
Third, how well the sample is mixed with the diluent after amplification.
For the former two points, CN115125129B and CN115141745B have given solutions.
However, although the prior application of "a constant temperature chromatography nucleic acid detecting apparatus and a detecting method thereof" has observed this problem with respect to "the degree of mixing with a diluent after amplification of a sample", it has been mainly solved from the aspect of "the height of a reaction tank".
However, the whole volume of the reaction tank is greatly limited by the design, and the detection capability of the constant temperature chromatographic nucleic acid detection device is limited to a certain extent.
Accordingly, the present application has been made keeping in mind the above problems.
Disclosure of Invention
The present application has been made in view of the above-described drawbacks of the prior art, and an object of the present application is to provide a dilution tube, a nucleic acid detecting apparatus, and a detecting method thereof.
The technical scheme of the application is as follows:
a dilution pipe comprising: an upper dilution pipe, a lower dilution pipe, a pipe plug and a diluent;
the upper dilution pipe is a hollow pipe body with an opening only at the bottom, a first annular bulge is arranged at the lower part of the lower dilution pipe, and threads are arranged on the outer surface of the first annular bulge; the upper dilution pipe is provided with an extrusion part;
the upper part of the lower dilution pipe is provided with a second annular bulge, and the outer surface of the second annular bulge is provided with threads;
a third annular bulge is arranged at the upper part of the lower dilution pipe, the radius of the third annular bulge is smaller than that of the second annular bulge, and the third annular bulge and the second annular bulge are arranged at intervals; the lower part of the lower dilution pipe is provided with a fourth annular bulge, and the inner surface of the fourth annular bulge is provided with threads;
wherein, lower dilution tunnel includes: the first space and the second space are communicated with each other up and down, and the first space is arranged above the second space; the first space and the second space are both cylindrical, and the cross-sectional area of the first space is larger than that of the second space; an opening is formed in the top of the first space, and an opening is formed in the bottom of the second space;
the top of the pipe plug is provided with a fifth annular bulge, and the periphery of the fifth annular bulge is provided with threads;
placing the diluent in the first space;
the threads of the first annular bulge are matched with the threads of the second annular bulge; the first annular bulge is arranged between the second annular bulge and the third annular bulge;
the fourth annular bulge of the lower dilution pipe is matched with the screw thread of the fifth annular bulge of the pipe plug, and the fifth annular bulge is screwed into the fourth annular bulge in a screw mode.
Further, the solution in the dilution tube is discharged from 200 to 300 mu L through x times of extrusion-release;
the inner diameter of the extrusion part of the upper dilution tube is ymm, and the diameter of the inner diameter of the second space is zmm; the values of y and z need to satisfy the following formula:
y=-0.25x 2 +3.25x-0.5；
z=0.005x 2 -0.015x+0.55。
further, the upper dilution tube is made of elastic materials.
Further, the upper dilution tube is made of plastic materials.
A nucleic acid detecting apparatus, comprising: the dilution tube, the surface of the nucleic acid detecting device body is provided with a sixth annular bulge, and the outer surface of the sixth annular bulge is provided with threads; the fourth annular bulge of the lower dilution tube of the dilution tube is matched with the screw thread of the sixth annular bulge, and the dilution tube is fixed on the nucleic acid detection device body in a sealing way through the fourth annular bulge and the sixth annular bulge.
Further, a non-medical purpose detection method comprising the steps of:
s1, adding a sampling liquid: sampling and adding the sampled solution into an upper opening of a vertical solution pipeline, and enabling the sampled solution to flow to a reaction cavity along the vertical solution pipeline, the liquid collecting cavity and a liquid collecting cavity-reaction cavity connecting pipeline;
s2, the dilution tube is mounted on the nucleic acid detecting device, so that the nucleic acid detecting device is kept in a sealed state;
s3, heating: starting a heating component to perform amplification reaction;
s4, adding a diluent:
first press-release:
extruding the upper dilution tube of the dilution tube for the first time, allowing part of the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity;
loosening the upper dilution tube of the dilution tube for the first time, and partially sucking back the solution, namely sucking back the solution in the reaction cavity to the liquid collecting cavity and the liquid collecting cavity-reaction cavity connecting pipeline along the first insertion groove;
second press-release:
extruding the upper dilution tube of the dilution tube for the second time, wherein part of the dilution liquid enters the nucleic acid detection device below, the dilution liquid enters the reaction cavity, and then the solution part in the reaction cavity is flushed into the dilution cavity;
releasing the upper dilution tube of the dilution tube for the second time, and partially sucking back the solution, namely sucking back the solution in the reaction cavity to the liquid collecting cavity and the liquid collecting cavity-reaction cavity connecting pipeline along the first inserting groove 2041;
repeating the press-loosening operation until the x-th press:
extruding the upper dilution tube of the dilution tube for the x time, allowing the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing a solution part in the reaction cavity into the dilution cavity;
the release for the x time can not be carried out in the back suction;
wherein x is 8 or 9 or 10;
and S5, enabling the solution to flow onto the test strip from the capillary drainage groove, and further obtaining a detection result.
The application has the beneficial effects that:
first, the basic idea of the application consists in a methodology: when the diluent in the diluent pipe enters the nucleic acid detection device, the diluent pushes the solution in the reaction cavity to flow to the diluent cavity and then to the chromatographic test paper when the diluent is extruded each time; the solution was refluxed each time it was released; after 8-10 times, the solution in the reaction cavity is continuously pushed and refluxed by the diluent, and the solution in the reaction cavity can be fully stirred in the process, so that the concentration of the amplified product is kept in a proper range, and the detection result of the chromatographic test paper is further ensured.
Second, a second application point of the present application is that:
the solution in the dilution tube is 200-300 mu L, and the solution in the dilution tube is discharged after x times of extrusion-release;
the inner diameter of the extrusion part of the upper dilution tube is ymm, and the diameter of the inner diameter of the second space is zmm; the values of y and z need to satisfy the following formula:
y=-0.25x 2 +3.25x-0.5；
z=0.005x 2 -0.015x+0.55。
the two indexes of the inner diameter are key indexes for ensuring extrusion for 8-10 times. The device and the method of the application have specific technical characteristics directly, and the device and the method meet the requirement of singleness.
Drawings
The application is described in further detail below in connection with the embodiments in the drawings, but is not to be construed as limiting the application in any way.
FIG. 1 is a schematic three-dimensional design of the dilution pipe of example 1.
FIG. 2 is a schematic cross-sectional design of the dilution pipe of example 1.
Fig. 3 is a schematic view of the installation process of the dilution pipe of example 1.
Fig. 4 is an actual view of the dilution pipe of example 2 in an assembled state.
FIG. 5 is a schematic cross-sectional view of a nucleic acid detecting apparatus according to example 2.
The reference numerals in fig. 1-5 are illustrated as follows:
an upper dilution pipe 101, a lower dilution pipe 102, a plug 103, a first space 1021, a second space 1022;
a vertical solution pipeline 201, a liquid collecting cavity 202, a liquid collecting cavity-reaction cavity connecting pipeline 203, a reaction cavity 204, a reaction cavity-dilution cavity connecting structure 205, a dilution cavity 206 and chromatographic test paper 207;
capillary drainage groove 2061;
a first insertion groove 2041, a freeze-dried ball stopper rod 2042;
ventilation holes 208 and a filter screen 209.
Detailed Description
The objects, technical solutions and advantages of the present application will become more apparent by the following detailed description of the present application with reference to the accompanying drawings. It should be understood that the description is only illustrative and is not intended to limit the scope of the application. In addition, in the following description, descriptions of well-known structures and techniques are omitted so as not to unnecessarily obscure the present application.
Example 1 monomer design of dilution tube
< structural design of dilution tube >
As shown in fig. 1 and 2, a dilution pipe 100 includes: an upper dilution pipe 101, a lower dilution pipe 102, a plug 103, and a diluent;
the upper dilution pipe 101 is constructed as follows: the upper dilution pipe is a hollow pipe body with an opening only at the bottom, a first annular bulge is arranged at the lower part of the lower dilution pipe, and threads are arranged on the outer surface of the first annular bulge; the upper dilution pipe 101 is of hose design;
the lower dilution tunnel 102 is constructed as follows: the upper part of the lower dilution pipe is provided with a second annular bulge, and the outer surface of the second annular bulge is provided with threads;
a third annular bulge is arranged at the upper part of the lower dilution pipe, the radius of the third annular bulge is smaller than that of the second annular bulge, and the third annular bulge and the second annular bulge are arranged at intervals;
the lower part of the lower dilution pipe 102 is provided with a fourth annular bulge, and the inner surface of the fourth annular bulge is provided with threads;
the lower dilution tube comprises: a first space 1021 and a second space 1022 which are communicated with each other up and down are obtained, and the first space is arranged above the second space; the first space and the second space are both cylindrical, and the cross-sectional area of the first space is larger than that of the second space;
an opening is arranged at the top of the first space (namely, an opening arranged at the top of the lower dilution pipe), and an opening is arranged at the bottom of the second space (namely, an opening arranged at the bottom of the lower dilution pipe);
a fifth annular protrusion is arranged at the top of the pipe plug 103, and threads are arranged at the periphery of the fifth annular protrusion;
the diluent is placed in the first space.
Matching relation:
the threads of the first annular bulge are matched with the threads of the second annular bulge; the first annular bulge is arranged between the second annular bulge and the third annular bulge;
the fourth annular projection of the lower dilution pipe 102 is fitted with the thread of the fifth annular projection of the plug 103, i.e. the fifth annular projection is screwed into the interior of the fourth annular projection.
Example 2 method of Using dilution tube and design emphasis ]
The application scene of the dilution tube is still CN115125129B, a constant temperature chromatographic nucleic acid detection device;
as shown in fig. 4 and 5, a nucleic acid detecting apparatus includes: the dilution tube, nucleic acid detecting body of example 1 (apparatus that may be "CN115125129B" or "CN 115141745B");
the nucleic acid detecting body includes: a solution flowing area, a heating component and a chromatographic test paper detecting area; the solution flowing region, the heating member and the chromatographic test paper detecting region are all arranged inside the nucleic acid detecting device.
1) A solution flow area comprising: a vertical solution pipeline 201, a liquid collecting cavity 202, a liquid collecting cavity-reaction cavity connecting pipeline 203, a reaction cavity 204, a reaction cavity-dilution cavity connecting structure 205 and a dilution cavity 206 which are sequentially communicated; the bottom surface of the reaction chamber 204 is lower than the bottom surface of the liquid collecting chamber-reaction chamber connecting line 203, and the bottom surface of the dilution chamber 206 is higher than the bottom surface of the reaction chamber 204.
The reaction chamber-dilution chamber connection configuration 205 is: the combined design of the U-shaped opening and the diversion canal is that the diversion canal (rectangular opening) is arranged at the bottom of the U-shaped opening, and the size of the diversion canal meets the following requirements: the ratio of the width of the guide channel to the width of the U-shaped opening is [0.2,0.4], and the aspect ratio of the guide channel is [0.85,1.2].
A first insertion groove 2041 and a freeze-drying ball limiting rod 2042 are inserted into the reaction cavity 204; the freeze-dried ball limiting rod 2042 is used for placing freeze-dried balls;
a capillary drainage groove 2061 is inserted into the dilution chamber 206; the liquid is drained onto the chromatographic strip by the action of the capillary drainage groove 2061.
In addition, the surface of the vertical solution channel 201, which is open at the upper part and protrudes from the nucleic acid detecting apparatus body, has a sixth annular protrusion, and the outer surface of the sixth annular protrusion is provided with a screw thread (the fourth annular protrusion of the lower part of the lower dilution tube 102 is adapted to the screw thread of the sixth annular protrusion of the lower nucleic acid detecting apparatus).
2) A heating member (which heats only the reaction chamber) is provided below the reaction chamber 204 to satisfy the nucleic acid detection amplification procedure.
3) A chromatographic test strip detection zone comprising: chromatographic strip 207; the chromatographic test paper 207 is inside the nucleic acid detecting device, the height of the end part of the chromatographic test paper 207 is higher than that of the dilution cavity 206, and the end part of the chromatographic test paper 207 connected with the dilution cavity 206 adopts a Z-shaped structure. The upper part of the other end of the nucleic acid detection device, which corresponds to the chromatographic test paper, is provided with a display window so as to check the result of the chromatographic test paper.
4) And an air vent 208 is also arranged, and a filter screen 209 is arranged in the middle of the air vent 208. The vent communicates with the solution flow area.
5) The number of solution flow areas may be plural.
For the problems mentioned in the background art: mixing uniformity.
The solution provided by the application is as follows:
a. as shown in fig. 3 and 4, in use, the dilution tube is unscrewed and the fourth annular projection on the lower part of the lower dilution tube 102 is rotatably connected to the sixth annular projection of the lower nucleic acid detecting apparatus; since the dilution liquid is placed in the first space, and the holes (cross sections) of the second space below are small, the liquid does not flow downwards (the solution is ensured not to be leaked into the nucleic acid detecting device below at will);
b. the core design of the dilution tube is as follows:
first press-release:
extruding the upper dilution tube of the dilution tube for the first time, allowing part of the dilution liquid to enter the nucleic acid detection device below, allowing the dilution liquid to enter the reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity 206;
the upper dilution tube of the dilution tube is loosened for the first time, and the solution is partially sucked back, namely, the solution in the reaction cavity is sucked back to the liquid collecting cavity 202 and the liquid collecting cavity-reaction cavity connecting pipeline 203 along the first inserting groove 2041;
second press-release:
extruding the upper dilution tube of the dilution tube for the second time, allowing part of the dilution liquid to enter the nucleic acid detection device below, allowing the dilution liquid to enter the reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity 206;
the upper dilution tube of the dilution tube is loosened for the second time, and the solution is partially sucked back, namely, the solution in the reaction cavity is sucked back to the liquid collecting cavity 202 and the liquid collecting cavity-reaction cavity connecting pipeline 203 along the first inserting groove 2041;
repeating the press-loosening operation until the x-th press:
extruding the upper dilution tube of the dilution tube for the x time, allowing the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing a solution part in the reaction cavity into a dilution cavity 206;
the x-th release will not cause the back suction (i.e. "the remaining solution volume after the x-1 th press-release" < "the solution volume of a single press", the x-th press will press the remaining dilution liquid into the nucleic acid detecting device, and at this time, the back suction will not occur).
The mixing effect is very good when x is 8-10 times.
That is, the past ideas are: the diluent is flushed from the diluent tube once and flows into the reaction cavity and then into the diluent cavity. However, the present application is different:
when the reaction chamber is squeezed each time, the diluent pushes the solution in the reaction chamber to flow to the dilution chamber and then to the chromatographic test paper; the solution was refluxed each time it was released; after 8-10 times, the solution in the reaction cavity is continuously pushed and refluxed by the diluent, and the solution in the reaction cavity can be fully stirred in the process, so that the concentration of the amplified product is kept in a proper range, and the detection result of the chromatographic test paper is further ensured.
The solution in the dilution tube is generally 200-300. Mu.L, and the key design of the above-mentioned "x is 8-10 times" is that:
A. the inner diameter of the squeeze portion provided for the upper dilution pipe 101 (which affects the volume of flow per squeeze) is set to ymm, and the inner diameter of the second space 1022 (which affects the volume of flow per reflux) is set to zmm; the values of y and z need to satisfy the following formula:
B. y=-0.25x 2 +3.25x-0.5
C. z=0.005x 2 -0.015x+0.55
the whole design process of the application is as follows:
s1, adding a sampling liquid: sampling and adding the sampled solution into an upper opening of the vertical solution pipeline 201, and flowing the sampled solution to the reaction cavity 204 along the vertical solution pipeline 201, the liquid collecting cavity 202 and the liquid collecting cavity-reaction cavity connecting pipeline 203;
s2, the dilution tube is mounted on the nucleic acid detecting device, so that the nucleic acid detecting device is kept in a sealed state;
s3, heating: starting a heating component to perform amplification reaction, wherein the solution after the reaction cavity 204 is ended is called mixed solution;
s4, adding a diluent:
first press-release:
extruding the upper dilution tube of the dilution tube for the first time, allowing part of the dilution liquid to enter the nucleic acid detection device below, allowing the dilution liquid to enter the reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity 206;
the upper dilution tube of the dilution tube is loosened for the first time, and the solution is partially sucked back, namely, the solution in the reaction cavity is sucked back to the liquid collecting cavity 202 and the liquid collecting cavity-reaction cavity connecting pipeline 203 along the first inserting groove 2041;
second press-release:
extruding the upper dilution tube of the dilution tube for the second time, allowing part of the dilution liquid to enter the nucleic acid detection device below, allowing the dilution liquid to enter the reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity 206;
the upper dilution tube of the dilution tube is loosened for the second time, and the solution is partially sucked back, namely, the solution in the reaction cavity is sucked back to the liquid collecting cavity 202 and the liquid collecting cavity-reaction cavity connecting pipeline 203 along the first inserting groove 2041;
repeating the press-loosening operation until the x-th press:
extruding the upper dilution tube of the dilution tube for the x time, allowing the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing a solution part in the reaction cavity into a dilution cavity 206;
the x-th loosening can not occur back suction.
And S5, enabling the solution to flow onto the test strip from the capillary drainage groove 1026, and further obtaining a detection result.
The above examples are provided for convenience of description of the present application and are not to be construed as limiting the application in any way, and any person skilled in the art will make partial changes or modifications to the application by using the disclosed technical content without departing from the technical features of the application.
Claims (7)
1. A dilution pipe, comprising: an upper dilution pipe, a lower dilution pipe, and a diluent;
the upper dilution pipe is a hollow pipe body with an opening only at the bottom, a first annular bulge is arranged at the lower part of the lower dilution pipe, and threads are arranged on the outer surface of the first annular bulge; the upper dilution pipe is provided with an extrusion part;
the upper part of the lower dilution pipe is provided with a second annular bulge, and the outer surface of the second annular bulge is provided with threads;
a third annular bulge is arranged at the upper part of the lower dilution pipe, the radius of the third annular bulge is smaller than that of the second annular bulge, and the third annular bulge and the second annular bulge are arranged at intervals; the lower part of the lower dilution pipe is provided with a fourth annular bulge, and the inner surface of the fourth annular bulge is provided with threads;
wherein, lower dilution tunnel includes: the first space and the second space are communicated with each other up and down, and the first space is arranged above the second space; the first space and the second space are both cylindrical, and the cross-sectional area of the first space is larger than that of the second space; an opening is formed in the top of the first space, and an opening is formed in the bottom of the second space;
placing the diluent in the first space;
the threads of the first annular bulge are matched with the threads of the second annular bulge; the first annular protrusion is disposed between the second annular protrusion and the third annular protrusion.
2. A dilution tunnel according to claim 1, further comprising: a pipe plug; the top of the pipe plug is provided with a fifth annular bulge, and the periphery of the fifth annular bulge is provided with threads; the fourth annular bulge of the lower dilution pipe is matched with the screw thread of the fifth annular bulge of the pipe plug, and the fifth annular bulge is screwed into the fourth annular bulge in a screw mode.
3. A dilution tunnel according to claim 1, wherein the solution in the dilution tunnel is discharged at 200 to 300 μl by x times of "squeeze-release";
the inner diameter of the extrusion part of the upper dilution tube is ymm, and the diameter of the inner diameter of the second space is zmm; the values of y and z need to satisfy the following formula:
y=-0.25x 2 +3.25x-0.5；
z=0.005x 2 -0.015x+0.55。
4. a dilution tunnel according to claim 1, wherein the upper dilution tunnel is made of an elastic material.
5. A dilution tunnel according to claim 1, wherein the upper dilution tunnel is made of plastic material.
6. A nucleic acid detecting apparatus, comprising: the dilution tube according to claim 1, 3, 4 or 5, wherein the surface of the nucleic acid detecting apparatus body is provided with a sixth annular protrusion, and the outer surface of the sixth annular protrusion is provided with threads; the fourth annular bulge of the lower dilution tube of the dilution tube is matched with the screw thread of the sixth annular bulge, and the dilution tube is fixed on the nucleic acid detection device body in a sealing way through the fourth annular bulge and the sixth annular bulge.
7. A detection method of a nucleic acid detecting apparatus, which is a non-medical object detection method, the nucleic acid detecting apparatus being the nucleic acid detecting apparatus according to claim 6;
the method is characterized by comprising the following steps of:
s1, adding a sampling liquid: sampling and adding the sampled solution into an upper opening of a vertical solution pipeline, and enabling the sampled solution to flow to a reaction cavity along the vertical solution pipeline, the liquid collecting cavity and a liquid collecting cavity-reaction cavity connecting pipeline;
s2, the dilution tube is mounted on the nucleic acid detecting device, so that the nucleic acid detecting device is kept in a sealed state;
s3, heating: starting a heating component to perform amplification reaction;
s4, adding a diluent:
first press-release:
extruding the upper dilution tube of the dilution tube for the first time, allowing part of the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing part of the solution in the reaction cavity into the dilution cavity;
loosening the upper dilution tube of the dilution tube for the first time, and partially sucking back the solution, namely sucking back the solution in the reaction cavity to the liquid collecting cavity and the liquid collecting cavity-reaction cavity connecting pipeline along the first insertion groove;
second press-release:
extruding the upper dilution tube of the dilution tube for the second time, wherein part of the dilution liquid enters the nucleic acid detection device below, the dilution liquid enters the reaction cavity, and then the solution part in the reaction cavity is flushed into the dilution cavity;
releasing the upper dilution tube of the dilution tube for the second time, and partially sucking back the solution, namely sucking back the solution in the reaction cavity to the liquid collecting cavity and the liquid collecting cavity-reaction cavity connecting pipeline along the first inserting groove 2041;
repeating the extrusion releasing operation until the x-th extrusion;
extruding the upper dilution tube of the dilution tube for the x time, allowing the dilution liquid to enter a nucleic acid detection device below, allowing the dilution liquid to enter a reaction cavity, and then flushing a solution part in the reaction cavity into the dilution cavity;
the release for the x time can not be carried out in the back suction;
wherein x is 8 or 9 or 10;
and S5, enabling the solution to flow onto the test strip from the capillary drainage groove, and further obtaining a detection result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311453962.8A CN117165416B (en) | 2023-11-03 | 2023-11-03 | Dilution tube, nucleic acid detection device and detection method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311453962.8A CN117165416B (en) | 2023-11-03 | 2023-11-03 | Dilution tube, nucleic acid detection device and detection method thereof |
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| Publication Number | Publication Date |
|---|---|
| CN117165416A true CN117165416A (en) | 2023-12-05 |
| CN117165416B CN117165416B (en) | 2024-01-26 |
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| CN202311453962.8A Active CN117165416B (en) | 2023-11-03 | 2023-11-03 | Dilution tube, nucleic acid detection device and detection method thereof |
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Citations (15)
| Publication number | Priority date | Publication date | Assignee | Title |
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