Nucleic acid detection device

Abstract

The nucleic acid detection device comprises an extraction end and a detection end which is communicated with the extraction end and is used for assembling an amplification reactant, the extraction end comprises a cracking part for assembling a cracking solution, a centrifugal part communicated with the cracking part, a filter membrane arranged between the cracking part and the centrifugal part, and a drawing plate movably inserted into the bottom end of the centrifugal part in the horizontal direction; an inner cavity of the detection end is provided with a flow measuring pipe communicated with the centrifugal part; wherein after a nucleic acid sample is subjected to reaction cracking in the cracking part, the nucleic acid sample after the reaction is extracted and collected by the filtering membrane through centrifugation, the nucleic acid sample is placed in the centrifugal part, and the drawing plate is drawn out in the horizontal direction, so that liquid enters the detection end along the flow measuring pipe; according to the utility model, nucleic acid extraction and isothermal amplification are combined, so that the detection time is greatly shortened, the application range is wide, the cost is low, meanwhile, the professional requirement on personnel is not high, and the result is visual and convenient.

Description

Technical Field

[0001] The utility model relates to the field of medical devices, and in particular to a nucleic acid detection device. Background Art

[0002] With the development of biotechnology, the detection and analysis of nucleic acids are increasingly being used for various detection purposes, including medical diagnosis, forensic identification, food testing, academic research, etc. Usually, the samples used for nucleic acid analysis are trace amounts, so in order to ensure the sensitivity of the detection, it is generally necessary to amplify the gene fragments containing the target nucleic acid molecules, and then use them for qualitative and quantitative detection, and then detect the amplified products or monitor the changes during the amplification process to determine whether the sample contains the target sequence. However, the process operation of the existing detection and amplification technology is very complicated and needs to be completed step by step on multiple devices. If there are many or even tens of thousands of samples to be tested, a lot of manpower and material resources need to be invested to share the work of each step, which undoubtedly increases the cost and detection cycle, and the cumbersome, single and boring steps are prone to operational errors. And as all fields of the biological industry pursue speed and automation, the limitations of traditional DNA extraction methods are becoming more and more obvious. The traditional polymerase chain reaction consists of a cycle of high-temperature denaturation, low-temperature annealing and temperature extension. This method has low cost, but takes a long time. It takes about 2 hours to amplify 40 cycles. The other method is probe-based PCR reaction. This method can shorten the detection time, but the probe synthesis is expensive, so the cost is higher. The traditional PCR amplification technology not only requires expensive instruments, but also takes a long time to amplify. What's more, in some remote areas, due to the shortage of many instruments and equipment, some samples cannot be amplified in time. If the test is carried out for a long time, it will cause problems such as sample damage, thus delaying further testing. Nowadays, many kits are used for extraction and amplification. Although the operation steps of the kit are simple, it also takes a long time. In amplification, the common amplification methods are ordinary PCR technology, real-time fluorescence quantitative PCR technology and digital PCR technology. Ordinary PCR amplification technology takes a long time and requires subsequent detection by gel electrophoresis, and the operation is relatively cumbersome. Real-time fluorescence quantitative PCR and digital PCR require relatively expensive reagents and instruments, and have high requirements for operators. They are not suitable for instant detection of nucleic acid samples in public places such as customs, homes, and schools, which is extremely inconvenient.

[0003] Therefore, it is necessary to provide a new technical solution to overcome the above-mentioned defects. Utility Model Content

[0004] The purpose of the utility model is to provide a nucleic acid detection device that can effectively solve the above-mentioned technical problems.

[0005] In order to achieve the purpose of this utility model, the following technical solutions are adopted:

[0006] A nucleic acid detection device, characterized in that it comprises an extraction end, a detection end connected to the extraction end and used to assemble an amplification reactant; the extraction end comprises a lysis portion with a lysis solution, a centrifugal portion connected to the lysis portion, a filter membrane arranged between the lysis portion and the centrifugal portion, and a pull-out plate movably connected to the bottom end of the centrifugal portion in a horizontal direction; the inner cavity of the detection end is provided with a flow tube for communicating with the centrifugal portion;

[0007] After the nucleic acid sample is lysed in the lysing part, the filter membrane extracts and collects the nucleic acid sample after the reaction and places it in the centrifugal part through centrifugation, and the pull-out plate is pulled out in the horizontal direction so that the liquid enters the detection end along the flow tube.

[0008] Furthermore, the extraction end is cylindrical in shape, the top of the lysis part is provided with a liquid inlet for the sample to flow in, and the liquid inlet is detachably provided with a closing cover; wherein the lysis part is detachably connected to the centrifugal part.

[0009] Furthermore, the shape of the detection end is conical, and the flow measuring tube is arranged to fit the inner wall of the detection end.

[0010] Furthermore, a partition is provided at one end of the flow tube close to the centrifugal part, and a drainage channel for communicating with the flow tube is provided on the partition; wherein the diameter of the drainage channel at one end close to the centrifugal part is greater than the diameter at one end away from the centrifugal part.

[0011] Furthermore, the flow tube is a hollow cylindrical structure, and the diameter of the drainage channel at one end away from the centrifugal part is equal to the diameter of the flow tube.

[0012] Furthermore, an end of the centrifugal part close to the detection end is provided with an opening for horizontal insertion or withdrawal of the pull-out plate, and an inner wall of the centrifugal part is provided with a guide groove for horizontal movement of the pull-out plate, and the guide groove is communicated with the opening.

[0013] Furthermore, a limit block is provided on the pull-out plate, and a limit groove cooperating with the limit block is provided in the guide groove.

[0014] Furthermore, a closing strip for plugging with the pull-out plate surrounds the circumference of the opening, wherein the closing strip is arranged on the outer wall of the centrifugal part.

[0015] Furthermore, the extraction end and the detection end are both made of transparent plastic material.

[0016] Furthermore, the cross-sectional length of the pulling plate is greater than the cross-sectional diameter of the extraction end.

[0017] Beneficial effects: The utility model combines nucleic acid extraction with isothermal amplification, which greatly shortens the detection time, has a wide range of applications, and is low in cost. At the same time, it does not require high professionalism of personnel, and the results are intuitive and convenient. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. They are used together with the embodiments of the present invention to explain the present invention and do not constitute a limitation to the present invention.

[0019] Figure 1 It is an application diagram of the utility model.

[0020] Figure 2 This is an exploded view of the utility model

[0021] Figure 3 It is the first structural schematic diagram of the utility model.

[0022] Figure 4 It is a second structural schematic diagram of the utility model.

[0023] Figure 5 It is a cross-sectional view of the utility model.

[0024] Figure 6 This is a schematic diagram of the structure of the pull-out plate of the utility model

[0025] In the figure: 1. extraction end, 2. detection end, 3. filter membrane, 4. centrifugal part, 5. lysis part, 6. pull-out plate, 7. liquid inlet, 8. partition, 9. drainage channel, 10. limiting groove, 11. closing strip, 12. closing cover, 13. guide groove, 14. limiting block, 15. flow tube, 16. opening. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solutions and advantages of the embodiments of the utility model clearer, the technical solutions in the embodiments of the utility model will be clearly and completely described below. Obviously, the described embodiments are partial embodiments of the utility model, rather than all embodiments.

[0027] In the description of the present utility model, it should be understood that the terms "center", "lateral", "longitudinal", "front", "rear", "left", "right", "up", "down", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the scope of protection of the present utility model. When a component is referred to as being "fixed to" another component, it may be directly on the other component or there may also be a centered component. When a component is considered to be "connected" to another component, it may be directly connected to the other component or there may be a centered component at the same time. When a component is considered to be "set on" another component, it may be directly set on the other component or there may be a centered component at the same time. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only.

[0028] like Figures 1 to 6 As shown, a nucleic acid detection device comprises an extraction end 1, and a detection end 2 connected with the extraction end 1 and used for assembling an amplification reactant; the extraction end 1 comprises a lysis portion 5 with a lysis solution, a centrifugal portion 4 connected with the lysis portion 5, a filter membrane 3 arranged between the lysis portion 5 and the centrifugal portion 4, and a pull-out plate 6 movably plugged in the horizontal direction with the bottom end of the centrifugal portion 4; the inner cavity of the detection end 2 is provided with a flow tube 15 for communicating with the centrifugal portion 4; wherein, after the nucleic acid sample is reacted and lysed in the lysis portion 5, the filter membrane 3 extracts and collects the reacted nucleic acid sample and places it in the centrifugal portion 4 through centrifugation, and the pull-out plate 6 is pulled out in the horizontal direction, so that the liquid enters the detection end 2 along the flow tube 15 to cause an amplification reaction.

[0029] Specifically, Figures 1 to 6As shown, the extraction end 1 and the detection end 2 are both made of transparent plastic material, the extraction end 1 is cylindrical in shape, the top of the lysis portion 5 is provided with a liquid inlet 7 for the sample to flow in, and the liquid inlet 7 is detachably provided with a closing cover 12, wherein the lysis portion 5 is detachably connected to the centrifugal portion 4; in this embodiment, the filter membrane 3 is arranged at the bottom end of the lysis portion 5, and the lysis portion 5 is inserted into the centrifugal portion 4 and screwed to the centrifugal portion 4, the cross-sectional shape of the closing cover 12 is T-shaped and the material of the closing cover 12 is plastic material, the end of the closing cover 12 with a shorter diameter is used for plugging with the liquid inlet 7, and a sealing ring is arranged on the end of the closing cover 12 with a shorter diameter, and the plastic material has a deformation feature, through the deformation feature and the sealing ring They cooperate with each other. On the one hand, it is convenient for medical staff to insert the closing cover 12 into the liquid inlet 7, and on the other hand, it can effectively enhance the airtightness of the connection between the closing cover 12 and the liquid inlet 7. In actual use, medical staff put the nucleic acid sample into the lysis part 5 through the liquid inlet 7, and the closing cover 12 closes the liquid inlet 7, so that the mixed liquid formed by mixing the nucleic acid sample with the lysis solution is centrifuged, and then the nucleic acid sample is filtered into the centrifugal part 4 through the filter membrane 3, and finally enters the detection end 2 for detection through the flow tube 15. The transparent plastic material makes it convenient for users to observe the internal conditions of the extraction end 1 and the detection end 2, and greatly improves the convenience and safety of nucleic acid detection, thereby improving the use effect of medical staff.

[0030] It is worth noting that the present embodiment is relatively small in size and is suitable for instant nucleic acid sample detection in public places such as customs, families, and schools. Only lysis solution and filter membrane 3 are needed for nucleic acid extraction. When further amplification detection is required, different lamp amplification reaction solutions can be designed according to the detection requirements, and the mixture can be mixed with primers and subjected to freeze-drying process to be converted into freeze-dried powder and placed in the inner cavity of the detection end 2. Since the lysis part 5 and the centrifugal part 4 are detachably connected, by disassembling the lysis part 5 and the centrifugal part 4, the user can place the freeze-dried powder formed by mixing the lamp amplification reaction solution with primers into the inner cavity of the detection end 2 through the centrifugal part 4. In actual use, the nucleic acid sample and the freeze-dried powder only need to be simply centrifuged, and then the detection end 2 can be placed in a thermos pot for reaction. The results are visible to the naked eye, which greatly shortens the detection time, is convenient and fast, and is convenient for subsequent operations.

[0031] It should be noted that loop-mediated isothermal amplification, referred to as LAMP, is a commonly used nucleic acid amplification technology nowadays. Four specific primers are designed for the six regions of the target gene, and under the action of chain displacement DNA polymerase, constant temperature amplification can be performed in the temperature range of 60-65°C. In the initial stage of amplification, due to the extension of the primers and the continuous chain displacement of the primary amplicon, a dumbbell-shaped reaction intermediate is formed. Compared with the traditional PCR reaction amplification mechanism, it is more complicated, but it greatly shortens the amplification time. 109-1010 nucleic acid amplification can be achieved within 15-60 minutes. The detection technology of LAMP itself is relatively low in cost compared to the RPA isothermal amplification technology.

[0032] like Figures 1 to 6 As shown, the shape of the detection end 2 is conical, and the flow tube 15 is arranged to fit the inner wall of the detection end 2, and a partition 8 is provided at the end of the flow tube 15 close to the centrifugal part 4, and a drainage channel 9 for communicating with the flow tube 15 is provided on the partition 8; wherein, the diameter of the drainage channel 9 close to the centrifugal part 4 is greater than the diameter of the end away from the centrifugal part 4, the flow tube 15 is a hollow cylindrical structure, and the diameter of the drainage channel 9 away from the centrifugal part 4 is equal to the diameter of the flow tube 15; in this embodiment, the drainage channel 9 is a hollow structure, and the drainage channel 9 connects the centrifugal part 4 and the flow tube 15; in actual use, the nucleic acid sample in the centrifugal part 4 flows into the drainage channel 9 and then enters the flow tube 15, and the drainage channel 9 can increase the flow rate of the nucleic acid sample, facilitate the nucleic acid sample to flow from the centrifugal part 4 into the flow tube 15, prevent the nucleic acid sample from being retained in the centrifugal part 4, avoid affecting the detection effect of the detection end 2, and effectively reduce the operation difficulty of this embodiment, and further effectively shorten the detection time.

[0033] like Figures 1 to 6As shown, an end of the centrifugal part 4 close to the detection end 2 is provided with an opening 16 for horizontal insertion or withdrawal of the pull-out plate 6, and an inner wall of the centrifugal part 4 is provided with a guide groove 13 for horizontal movement of the pull-out plate 6, the guide groove 13 and the opening 16 are interconnected, a limit block 14 is provided on the pull-out plate 6, and a limit groove 10 is provided in the guide groove 13 to cooperate with the limit block 14 and abut against it. The cross-sectional length of the pull-out plate 6 is greater than the cross-sectional diameter of the extraction end 1; in the present embodiment, the pull-out plate 6 is arranged at the bottom end of the centrifugal part 4, and is in close contact with the end of the drainage channel 9 with a larger diameter, and the end of the pull-out plate 6 close to the drainage channel 9 is in an arc shape that is in close contact with the centrifugal part 4, and the pull-out plate 6 is used to block and control the liquid flow; the limit block 14 and the pull-out plate 6 are an integrated structure, and the groove volume of the limit groove 10 is greater than the volume of the limit block 14, the limit block 14 is plugged into the limit groove 10 and the limit block 14 can move in the limit groove 10, and the pull-out plate 6 can move axially along the guide groove 13 at the bottom end of the centrifugal part 4 through the opening 16; in actual use, the user pulls out the pull-out plate 6 to the outside along the guide groove 13, so that the pull-out plate 6 and the centrifugal part 4, the nucleic acid sample flows into the drainage channel 9 through the gap, and the pull-out plate 6 can effectively control the liquid flow from the centrifugal part 4 to the drainage channel 9, and control the opening size of the drainage channel 9, so that the user can adjust the nucleic acid sample entering the detection end 2, which is convenient for the user to use. On the other hand, through the mutual cooperation between the limit block 14 and the limit groove 10, and the groove volume of the limit groove 10 is larger than the volume of the limit block 14, the limit groove 10 can be used as a moving track of the limit block 14 on the one hand, so that the limit block 14 can move through the limit groove 10 in this embodiment, and on the other hand, the moving range of the pull-out plate 6 is effectively limited to prevent the pull-out plate 6 from being pulled out too long and thus separated from the centrifugal part 4, thereby avoiding affecting the normal use of this embodiment.

[0034] To further optimize this embodiment, Figures 1 to 6 As shown, the opening 16 is surrounded by a closing strip 11 for plugging with the pull-out plate 6, wherein the closing strip 11 is arranged on the outer wall of the centrifugal part 4; in this embodiment, the closing strip 11 is made of rubber material, and the closing strip 11 can effectively enhance the stability of the connection between the opening 16 and the pull-out plate 6, so that the liquid in the pull-out plate 6 is blocked in the centrifugal part 4 through the rubber material, and effectively prevents the nucleic acid sample from being taken out or brought back by the pull-out plate 6 when the pull-out plate 6 moves, thereby avoiding affecting the detection result of the nucleic acid sample.

[0035] Working principle: put the nucleic acid sample into the lysis part 5 from the liquid inlet 7, so that the nucleic acid sample and the lysis solution are evenly mixed and shaken for at least 1 minute to fully react and lyse; the fully reacted nucleic acid sample is retained on the filter membrane 3 through the centrifugal operation, and is extracted into the centrifugal part 4 through the filter membrane 3, and the pull-out plate 6 is pulled out to control the liquid flow of the nucleic acid sample, so that the nucleic acid sample flows along the drainage channel 9 from the centrifugal part 4 to the inner cavity of the detection end 2, and the amplification primer in the inner cavity of the detection end 2 is mixed with the nucleic acid sample to cause an amplification reaction.

[0036] The standard parts used in the utility model can be purchased from the market, and special-shaped parts can be customized according to the description in the specification and the drawings. The specific connection methods of each part adopt the conventional means such as mature bolts, rivets, welding, etc. in the prior art. The machinery, parts and equipment all adopt the conventional models in the prior art, and the circuit connection adopts the conventional connection method in the prior art, which will not be described in detail here. The content not described in detail in this specification belongs to the prior art known to professional and technical personnel in this field.

[0037] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should fall within the scope of protection of the claims attached to the utility model.

Claims

1. A nucleic acid detection device, characterized in that: The invention comprises an extraction end (1), and a detection end (2) which is connected to the extraction end (1) and is used to hold an amplification reaction product; the extraction end (1) comprises a lysis portion (5) for holding a lysis solution, a centrifugal portion (4) which is connected to the lysis portion (5), a filter membrane (3) arranged between the lysis portion (5) and the centrifugal portion (4), and a pull-out plate (6) which is horizontally movably plugged into the bottom end of the centrifugal portion (4); the inner cavity of the detection end (2) is provided with a flow tube (15) which is used to communicate with the centrifugal portion (4); After the nucleic acid sample is lysed in the lysing section (5), centrifugation is performed so that the filter membrane (3) extracts and collects the nucleic acid sample after the reaction and places it in the centrifugal section (4). The pull-out plate (6) is pulled out in the horizontal direction so that the liquid flows along the flow tube (15) into the detection end (2) to cause an amplification reaction.

2. A nucleic acid detection device according to claim 1, characterized in that: The extraction end (1) is cylindrical in shape, and the top of the lysis section (5) is provided with a liquid inlet (7) for the sample to flow in, and the liquid inlet (7) is detachably provided with a sealing cover (12); wherein the lysis section (5) is detachably connected to the centrifugal section (4).

3. A nucleic acid detection device as claimed in claim 2, characterized in that: The detection end (2) is in the shape of a cone, and the flow measuring tube (15) is arranged to fit the inner wall of the detection end (2).

4. A nucleic acid detection device as claimed in claim 3, characterized in that: A partition plate (8) is provided at one end of the flow measuring tube (15) close to the centrifugal part (4), and a drainage channel (9) for communicating with the flow measuring tube (15) is provided on the partition plate (8); wherein the diameter of the drainage channel (9) at the end close to the centrifugal part (4) is greater than the diameter of the end away from the centrifugal part (4).

5. A nucleic acid detection device as claimed in claim 4, characterized in that: The flow tube (15) is a hollow cylindrical structure, and the diameter of the end of the drainage channel (9) away from the centrifugal part (4) is equal to the diameter of the flow tube (15).

6. A nucleic acid detection device as claimed in claim 5, characterized in that: An end of the centrifugal part (4) close to the detection end (2) is provided with an opening (16) for horizontal insertion or withdrawal of the pull-out plate (6), and an inner wall of the centrifugal part (4) is provided with a guide groove (13) for horizontal movement of the pull-out plate (6), and the guide groove (13) and the opening (16) are communicated with each other.

7. A nucleic acid detection device according to claim 6, characterized in that: A limit block (14) is provided on the pull-out plate (6), and a limit groove (10) is provided in the guide groove (13) for cooperating and abutting with the limit block (14).

8. A nucleic acid detection device according to claim 7, characterized in that: The opening (16) is surrounded by a closing strip (11) for plugging into the drawer plate (6), wherein the closing strip (11) is arranged on the outer wall of the centrifugal part (4).

9. A nucleic acid detection device according to claim 1, characterized in that: The extraction end (1) and the detection end (2) are both made of transparent plastic material.

10. A nucleic acid detection device according to claim 1, characterized in that: The cross-sectional length of the pulling plate (6) is greater than the cross-sectional diameter of the extraction end.

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