Micro-fluidic chip card box for PCR (Polymerase Chain Reaction) detection
By pre-embedding PCR amplification reagent in the microfluidic chip box and simplifying operation with a one-way soft valve and runner mechanism, the problems of high detection costs and complex operation in the prior art are solved, and convenient real-time fluorescence PCR detection is achieved.
Patent Information
- Application Number
- CN202421493200.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-27
- Publication Date
- 2025-07-25
- Estimated Expiration
- 2034-06-27
AI Technical Summary
The existing microfluidic chip cartridges have problems such as high detection cost, complex operation and insufficient flexibility in real-time fluorescence PCR detection, especially the single-chip method requires complex reagent preparation and a high-demand laboratory environment.
A microfluidic chip card box is designed, and the PCR amplification reagent is embedded in the cavity. Sample liquid is injected into the one-way soft valve and mixed. The mixed liquid is introduced into the microfluidic chip reaction chamber using a runner mechanism, simplifying operation and integrating the amplification and detection steps.
It reduces the work of users to prepare amplification reagents, is simple and convenient to operate, reduces costs, and improves the flexibility and efficiency of detection throughput.
Smart Images
Figure CN223150557U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the field of medical detection, in particular to a microfluidic chip cartridge for PCR detection. Background Art
[0002] Molecular diagnosis by nucleic acid in vitro amplification methods includes real-time fluorescence PCR (polymerase chain reaction), isothermal amplification, etc. Disease diagnosis is carried out by detecting specific gene fragments of pathogens, which has higher sensitivity and stronger specificity compared with antigen-antibody detection. Among them, the real-time fluorescence PCR method is even the gold standard for the detection of many pathogens.
[0003] At present, there are mainly two methods for real-time fluorescence PCR detection using the microfluidic chip method: an integrated cartridge and a single chip. Among them, the integrated cartridge method integrates the extraction reagent, amplification reagent, and detection chip in the cartridge. Users only need to add samples and place the cartridge in the detection device to achieve the detection of "sample in, result out". However, this method can only detect one sample at a time, and the detection cost is relatively high. The single chip method requires the work of extracting and preparing the amplification reagent outside the chip, adding the reagent to the chip, and then putting the chip on the machine for detection. This method is more flexible in detection. The number of samples or the number of detection targets can be increased by increasing the number of chips, and the cost is relatively low. However, the operation of preparing the reagent outside the chip is complex, and the requirements for the PCR laboratory environment are high, which also restricts the implementation of PCR detection from being fast and convenient.
[0004] Therefore, there is a need for a microfluidic chip cartridge for real-time fluorescence PCR detection that can not only reduce the complexity of reagent preparation for users, but also increase the flexibility of extraction quantity, improve the detection throughput, and better reduce the cost. Summary of the Utility Model
[0005] The embodiments of the utility model provide a microfluidic chip cartridge for PCR detection to solve at least one of the problems existing in the related art. To achieve this purpose, the utility model is realized through the following technical solutions.
[0006] The utility model provides a microfluidic chip cartridge for PCR detection, including: an anti-overflow soft plug, a cavity pre-embedded with PCR amplification reagent, a microfluidic chip, and a buckle cover. Among them: at least one reagent chamber is arranged in the cavity, at least one one-way soft valve is arranged on the anti-overflow soft plug, and the anti-overflow soft plug is used for sealing the cavity and injecting the extracted sample liquid into the corresponding reagent chamber through the one-way soft valve and then sealing; a flow channel mechanism is arranged at the bottom of the cavity, and the sample liquid mixed with the amplification reagent enters the reaction cavity of the microfluidic chip through the flow channel mechanism; the buckle cover is clamped with the flow channel mechanism and is used for clamping the microfluidic chip between the buckle cover and the flow channel mechanism.
[0007] Furthermore, the microfluidic chip cartridge for PCR detection further includes a cover body, and the cover body is detachably connected to the cavity.
[0008] Furthermore, the microfluidic chip cartridge for PCR detection further includes a pressing plate. The center of the pressing plate has a protrusion, and the center of the cover body has a hole. The protrusion passes through the hole in the center of the cover body to achieve a clearance fit.
[0009] Furthermore, one end of the at least one one-way soft valve facing the inside of the cavity is a flat normally closed end and is sealed with the corresponding reagent chamber around it, and the end facing the outside of the cavity is a generally circular normally open end.
[0010] Furthermore, a sealing convex edge is provided around the normally open end of each one-way soft valve.
[0011] Furthermore, liquid silicone is embedded inside the flow channel mechanism through a two-color mold process to form a plurality of flow channel silicone pads, and a flow channel convex edge is formed at the outer edge of the flow channel mechanism.
[0012] Furthermore, the sample inlet of the microfluidic chip corresponds to the position of the flow channel mechanism.
[0013] Furthermore, patterned bumps are formed at the bottom of the snap-on cover corresponding to the position of the flow channel mechanism, and the bumps correspond to the position of the sample inlet of the microfluidic chip.
[0014] Furthermore, the snap-on cover is detachably clamped to the flow channel mechanism.
[0015] The embodiments of the present utility model have the following beneficial effects:
[0016] (1) For the microfluidic chip cartridge for PCR detection provided by the present utility model, the PCR amplification reagent is pre-buried in the cartridge cavity. The user adds the extracted nucleic acid sample solution into the cavity to mix with the PCR amplification reagent, and then uses an external centrifugal device to passively introduce the mixed reagent into the reaction cavity of the microfluidic chip through centrifugation operation, and then PCR detection can be carried out in a supporting PCR detection instrument. For the microfluidic chip cartridge for PCR detection provided by the present utility model, by performing the nucleic acid extraction step outside the cartridge and pre-burying the PCR amplification reagent in the cartridge cavity, that is, the amplification and detection steps are integrated in the cartridge, the work of the user in preparing the amplification reagent is reduced, the operation is simple and convenient, and at the same time, the requirements for extraction are more flexible, the cost is better reduced, and it is convenient to improve the detection throughput.
[0017] (2) The microfluidic chip cartridge for PCR detection provided by the present utility model embeds liquid silicone into the flow channel mechanism at the bottom of the cavity through a two-color mold process, enabling the combination of hard plastic and soft rubber in the microchannel, which not only provides sealing but also protects the silicon substrate of the microfluidic chip from being crushed.
[0018] (3) The one-way soft valve provided on the anti-overflow soft plug for sealing the cartridge cavity utilizes the elastic sealing property of silicone, and one end facing the inside of the cavity is a flat normally closed end, while the end facing the outside of the cavity is a circularly shaped normally open end, allowing liquid to be injected from the outside while preventing the internal liquid from flowing out, and the liquids in each reagent chamber do not interfere with each other, which is convenient for liquid injection and achieves sealing.
[0019] (4) A sealing convex edge is provided around the normally open end of the anti-overflow soft plug, and a sealing strip is formed between the sealing convex edge and the pressing plate pressing on the anti-overflow soft plug, making the sealing performance of the anti-overflow soft plug better when the cover body is screwed on.
[0020] (5) Protrusions corresponding to each sampling port of the microfluidic chip are designed on the buckle cover for pressing the microfluidic chip, avoiding the phenomenon that the pressure applied to the sampling port is insufficient due to the large-area contact between the microfluidic chip and the buckle cover. The targeted contact reduces the contact area and increases the pressing force. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] The drawings forming a part of the present utility model are used to provide a further understanding of the present utility model. The schematic embodiments of the present utility model and their descriptions are used to explain the present utility model and do not constitute an improper limitation of the present utility model.
[0022] Figure 1 is the structural explosion diagram of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model;
[0023] Figure 2 is the structural assembly diagram of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model;
[0024] Figure 3 is the schematic cross-sectional view of the open cover state of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model;
[0025] Figure 4 is the bottom view, top view and side view of the anti-overflow soft plug of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model.
[0026] Reference Signs:
[0027] 1 - Cover body; 2 - Pressure plate; 3 - Anti - overflow soft plug; 4 - PCR amplification reagent; 5 - Cavity; 6 - Microfluidic chip; 7 - Cover buckle; 8 - Runner gasket; 9 - One - way soft valve; 10 - Sealing flange; 11 - Protrusion; 12 - Runner flange; 14 - Runner mechanism; 16 - Reagent chamber; 17 - Protrusion; 18 - Normally closed end; 19 - Normally open end. Detailed implementation mode
[0028] To make the objectives, technical solutions and advantages of the embodiments of the present utility model clearer, the following will elaborate on each implementation mode of the present utility model with reference to the accompanying drawings. However, those of ordinary skill in the art can understand that in each implementation mode of the present utility model, many technical details are provided for the reader to better understand the present application. However, even without these technical details and various changes and modifications based on the following implementation modes, the technical solutions claimed in the present application can still be achieved. The division of the following various embodiments is for the convenience of description and should not constitute any limitation on the specific implementation mode of the present utility model. The various embodiments can be combined and cross - referenced with each other on the premise of no contradiction.
[0029] The embodiment of the present utility model provides a microfluidic chip cartridge for PCR detection. Figures 1-3 Respectively are the structural explosion diagram, assembly diagram and cross - sectional schematic diagram of the open - cover state of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model. As Figure 1 And Figure 2 Shown, the microfluidic chip cartridge for PCR detection includes: cover body 1, pressure plate 2, anti - overflow soft plug 3, cavity 5 pre - embedded with PCR amplification reagent 4, microfluidic chip 6 and cover buckle 7. Cavity 5, microfluidic chip 6 and cover buckle 7 are the three main components of the cartridge. Cavity 5 is provided with one or more reagent chambers 16, and PCR amplification reagent 4, such as freeze - dried reagent, is pre - embedded in the reagent chamber 16. In this embodiment, the microfluidic chip 6 can be made of silicon and heat - resistant materials (such as glass, PC, PP, silica gel, etc.).
[0030] The anti - overflow soft plug 3 is used to seal the open end of the cavity 5 and inject the extracted sample to be tested into the cavity 5 through the anti - overflow soft plug 3. In this embodiment, the anti - overflow soft plug 3 can be made of silica gel and is hermetically bonded to the screw - cap type cavity 5 through silica gel glue. Figure 4 Are the bottom view, top view and side view of the anti - overflow soft plug of the microfluidic chip cartridge for PCR detection according to the embodiment of the present utility model. As Figure 4As shown, one or more one-way soft valves 9 are provided on the anti-overflow soft plug 3. The number and position of the one-way soft valves 9 match and correspond to the number and position of the reagent chambers 16 of the cavity 5. The end of the one-way soft valve 9 facing the inside of the cavity 5 is a flat normally closed end 18, and the periphery of the normally closed end 18 is tightly sealed with its corresponding reagent chamber 16. The end facing the outside of the cavity 5 is a circular-like normally open end 19. The extracted sample liquid is injected into each reagent chamber 16 through the normally open ends 19 of the respective one-way soft valves 9. By utilizing the elastic airtight property of silica gel and the specific structure of the one-way soft valve 9, liquid can be injected from the outside, the internal liquid cannot flow out, and the liquids in the respective reagent chambers 16 do not interfere with each other, facilitating liquid injection and achieving sealing at the same time.
[0031] In this embodiment, the cavity 5 can be a screw-cap type cavity, and the cover 1 can be a screw-type cover. The cover 1 is detachably connected to the cavity 5. As Figure 1 and Figure 2 shown, the center of the pressing plate 2 has a protrusion 17, and the center of the cover 1 has a hole. During assembly, the pressing plate 2 is placed on the anti-overflow soft plug 3. When the cover 1 is put on, the protrusion 17 passes through the hole in the center of the cover 1, and a clearance fit is achieved between the protrusion 17 and the hole in the center of the cover 1. Preferably, as Figure 3 shown, a sealing flange 10 is provided on the periphery of the normally open end 19 of the anti-overflow soft plug 3. A sealing strip is formed between the sealing flange 10 and the pressing plate 2 pressing on the anti-overflow soft plug 3, making the sealing performance of the anti-overflow soft plug 3 better when the cover 1 is screwed on.
[0032] As Figure 1 shown, a flow channel mechanism 14 is provided at the bottom of the cavity 5. The flow channel mechanism 14 is communicated with the reagent chambers 16 in the cavity 5. The sample liquid added through the normally open ends 19 of the anti-overflow soft plug 3 is mixed with the PCR amplification reagent 4 and then enters the reaction chamber of the microfluidic chip 6 through the flow channel mechanism 14. In this embodiment, liquid silicone is embedded inside the flow channel mechanism 14 through a two-color molding process to form a plurality of flow channel silicone pads 8, enabling the combination of hard plastic and soft rubber in the micro-channel, achieving sealing while protecting the silicon substrate of the microfluidic chip 6 from being damaged. As Figure 3 shown, a flow channel flange 12 is formed on the outer edge of the flow channel mechanism 14, which can achieve a better sealing effect with the silicon-based chip 6. The buckling cover 7 is snap-connected to the flow channel mechanism 14 for clamping the microfluidic chip 6 between the buckling cover 7 and the flow channel mechanism 14 of the cavity 2. Preferably, the buckling cover 7 is detachably snap-connected to the flow channel mechanism 14, facilitating the loading of the microfluidic chip. Among them, the sample inlet of the microfluidic chip 6 corresponds to the position of the flow channel mechanism 14, facilitating the sample liquid to enter the reaction chamber of the microfluidic chip 6 from the flow channel mechanism 14.
[0033] Preferably, as Figure 3As shown, the bottom of the buckle cover 7 is formed with patterned convex points 11 at the position corresponding to the flow channel mechanism 14, and the convex points 11 correspond to the position of the injection port of the microfluidic chip 6. The convex points 11 corresponding to each injection port of the microfluidic chip 6 are designed on the buckle cover 7 for pressing the microfluidic chip 6, so as to avoid the phenomenon that the microfluidic chip 6 and the buckle cover 7 are in contact with a large area and the pressure on the injection port is not applied in place, and the contact is targeted to reduce the contact area and increase the pressing force.
[0034] The sample loading process of the microfluidic chip cartridge for PCR detection is as follows: the screw-on cover 1 is lightly screwed on the open end of the screw-on cover button cavity 5; when in use, the screw-on cover 1 is first unscrewed, bringing out the pressure plate 2, and a pipette is used to penetrate the normally open end 19 of the anti-overflow soft plug 3 and extend into the reagent chamber 16 inside the screw-on cover button cavity 5 through the normally closed end 18, and a sample liquid is added. After the pipette is withdrawn, the normally closed end 18 of the anti-overflow soft plug 3 is closed, and the sample liquid is sealed in the reagent chamber 16 and dissolved with the freeze-dried reagent pre-embedded in the reagent chamber 16, and then the screw-on cover 1 is installed and tightened; it is placed in a centrifuge (1000-2000r / min) and centrifuged (30 seconds), and the fused sample to be tested is thrown into the reaction chamber of the microfluidic chip 6 to complete the sample loading process.
[0035] The microfluidic chip cartridge for PCR detection provided by the utility model pre-buries the PCR amplification reagent in the cartridge cavity. The user adds the extracted nucleic acid sample liquid into the cavity to mix with the PCR amplification reagent, and then uses an external centrifugal device to passively enter the mixed reagent into the reaction cavity of the microfluidic chip through centrifugal operation, and PCR detection can be performed in the matching PCR detection instrument. The microfluidic chip cartridge for PCR detection provided by the utility model performs the nucleic acid extraction step outside the cartridge, and pre-buries the PCR amplification reagent in the cartridge cavity, that is, the amplification and detection steps are integrated in the cartridge, which reduces the user's work of preparing the amplification reagent, is simple and convenient to operate, and is more flexible for the extraction requirements, better reduces the cost, and is convenient to improve the detection throughput.
[0036] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A microfluidic chip cartridge for PCR detection, characterized in that, Including: an anti-overflow soft plug (3), a cavity (5) pre-embedded with a PCR amplification reagent (4), a microfluidic chip (6), and a cover cap (7), wherein: at least one reagent chamber (16) is provided in the cavity (5), at least one one-way soft valve (9) is provided on the anti-overflow soft plug (3), and the anti-overflow soft plug (3) is used for sealing the cavity (5) and injecting the extracted sample liquid into the corresponding reagent chamber (16) through the one-way soft valve (9) and then sealing; a flow channel mechanism (14) is provided at the bottom of the cavity (5), the flow channel mechanism (14) communicates with each reagent chamber (16), and the sample liquid mixed with the PCR amplification reagent (4) enters the reaction chamber of the microfluidic chip (6) through the flow channel mechanism (14); the cover cap (7) is detachably clamped with the flow channel mechanism (14) and is used for clamping the microfluidic chip (6) between the cover cap (7) and the flow channel mechanism (14).
2. The microfluidic chip cartridge for PCR detection according to claim 1, wherein It further includes a cover body (1), and the cover body (1) is detachably connected to the cavity (5).
3. The microfluidic chip cartridge for PCR detection according to claim 2, wherein, It further includes a pressing plate (2), a protrusion (17) is provided at the center of the pressing plate (2), a hole position is provided at the center of the cover body (1), and the protrusion (17) passes through the hole position at the center of the cover body (1) to achieve clearance fit.
4. The microfluidic chip cartridge for PCR detection according to claim 3, wherein One end of each one-way soft valve (9) facing the inside of the cavity (5) is a flat normally-closed end (18) and its periphery is sealed with the corresponding reagent chamber (16), and one end facing the outside of the cavity (5) is a quasi-circular normally-open end (19).
5. The microfluidic chip cartridge for PCR detection according to claim 4, characterized in that, A sealing convex edge (10) is provided around the normally-open end (19) of each one-way soft valve (9).
6. The microfluidic chip cartridge for PCR detection according to claim 1, wherein Liquid silicone is embedded inside the flow channel mechanism (14) through a two-color mold process to form a plurality of flow channel silicone pads (8), and a flow channel convex edge (12) is formed at the outer edge of the flow channel mechanism (14).
7. The microfluidic chip cartridge for PCR detection according to claim 1, characterized in that, The sample inlet of the microfluidic chip (6) corresponds to the position of the flow channel mechanism (14).
8. The microfluidic chip cartridge for PCR detection according to claim 7, wherein Patterned bumps (11) are formed at the bottom of the cover cap (7) corresponding to the flow channel mechanism (14), and the bumps (11) correspond to the position of the sample inlet of the microfluidic chip (6).
9. The microfluidic chip cartridge for PCR detection according to claim 1, characterized in that, The cover cap (7) is detachably clamped with the flow channel mechanism (14).