Microfluidic card box for multiple detection of nucleic acid of pathogenic microorganisms
Through the design of the microfluidic card box, efficient and accurate distribution and reaction of multiple detection of pathogenic microbial nucleic acids is achieved, and the problems of uneven and cross-reaction of sample addition in the prior art are solved, and detection efficiency and accuracy are improved.
Patent Information
- Application Number
- CN202422137215.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-02
- Publication Date
- 2025-07-29
- Estimated Expiration
- 2034-09-02
AI Technical Summary
The existing multi-detection technology for pathogenic microbial nucleic acids requires multiple sets of experiments. The addition of samples is time-consuming and laborious, and it is difficult to ensure that the components are consistent, there are errors, and cross-reactions are difficult to avoid.
A microfluidic card box is designed, including a sample tube, a reaction base and a split assembly. The liquid is evenly distributed from bottom to top through the flow column and the overflow filter element to avoid bubble generation, and ensure the stability of the reaction box and liquid closure through the snap ring and leakage absorption tank to avoid cross-reaction.
Achieve one-step sample allocation to complete sample allocation, avoid cross-reactions, improve detection efficiency and accuracy, simplify operational processes, and ensure reaction consistency.
Smart Images
Figure CN223163422U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of biological detection, and specifically relates to a microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms. Background Technique
[0002] Multiplex detection of nucleic acids of pathogenic microorganisms is an efficient molecular detection technology that can simultaneously detect the nucleic acid sequences of multiple pathogenic microorganisms in the same reaction system. This technology has advantages such as high throughput, high sensitivity, and high specificity, and is widely used in fields such as pathogen diagnosis, screening, prevention, and control of infectious diseases. Multiplex detection of nucleic acids of pathogenic microorganisms is mainly based on the principles of nucleic acid amplification and detection. Commonly used technologies include multiplex polymerase chain reaction (PCR), multiplex isothermal amplification technology (ITA), multiplex nucleic acid detection technology based on the CRISPR / Cas system, and metagenomic sequencing (mNGS), etc.
[0003] In current multiplex detection technologies for nucleic acids of pathogenic microorganisms, generally, the sample is placed in a test tube, and after adding reaction reagents for reaction, it is then detected. However, in the actual detection process, it is usually necessary to set up a control group or perform detection with different types of reagents. Therefore, multiple groups of experiments are required. And the addition of samples is generally carried out by a dropper. Conducting multiple groups of experiments is time-consuming and laborious, and it is difficult to ensure that the amount of each group is consistent. Therefore, there are certain errors. Content of the Utility Model
[0004] In view of the deficiencies of the prior art, the utility model provides a microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms, which solves the problems existing in the existing detection technology.
[0005] To achieve the above objectives, the utility model is realized through the following technical solutions: A microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms, comprising:
[0006] A sample tube for accommodating a nucleic acid sample of a pathogenic microorganism to be detected or a reagent for sample treatment;
[0007] A reaction base for shunting and reacting the sample in the sample tube;
[0008] The top of the sample tube is connected with a sealing cover through a flexible connector on one side, and a valve is arranged at the bottom of the sample tube. The reaction base is connected to the bottom end of the sample tube in a plug-in manner and pushes open the valve to make the liquid in the sample tube flow out. A shunting component for uniformly shunting and reacting the liquid flowing out of the sample tube is arranged inside the reaction base.
[0009] Preferably, the reaction base includes an upper shell and a bottom cover, and the upper shell and the bottom cover are fixedly connected by bolts. A card seat for clamping the sample tube is arranged in the middle of the top of the upper shell.
[0010] Preferably, a plurality of anti-overflow holes are recessed in the periphery of the top of the upper shell, and anti-overflow filter elements that expand after absorbing liquid are plugged in the anti-overflow holes.
[0011] Preferably, the shunt assembly includes:
[0012] A hollow guide post, fixedly connected through the center of the upper shell, used to push open the valve, and the top of the hollow guide post is provided with an opening;
[0013] Fixing pieces, a plurality of which are provided and evenly distributed at the bottom of the bottom cover;
[0014] A diversion post, fixedly connected to one side of the bottom of the fixing piece;
[0015] A shunt tube, with two ends respectively connected to the hollow guide post and the diversion post;
[0016] A drainage branch pipe, fixedly connected through one side of the diversion post at one end, and the other end of the drainage branch pipe is communicated with the bottom of the side of the anti-overflow hole.
[0017] Preferably, a pressing plate is detachably arranged at the bottom of the reaction base, a plurality of reaction boxes are evenly penetrated in the circumferential direction inside the pressing plate, and the pressing plate presses the reaction boxes against the bottom of the fixing piece to achieve sealing. Nucleic acid amplification reagents and primer probes are preset in the reaction boxes before detection.
[0018] Preferably, a snap ring is rotatably sleeved on the outside of the upper shell, a limiting strip is fixedly connected to the side of the upper shell and penetrates through the snap ring, and a limiting groove for restricting the rotation angle of the limiting strip is arranged inside the snap ring.
[0019] Preferably, a plurality of clamping strips are evenly arranged on the outside of the pressing plate, and a plurality of clamping grooves for rotatably clamping the clamping strips are evenly arranged on the snap ring, so as to connect the pressing plate to the bottom of the upper shell.
[0020] Preferably, a leakage absorption groove is recessed at the top of the reaction base and around the card seat, and a sponge ring is arranged in the leakage absorption groove.
[0021] The utility model provides a microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms. Compared with the prior art, the following beneficial effects are achieved:
[0022] 1. The microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acid only requires one-step sample loading and assembly actions to complete the injection of samples and the equal distribution into multiple reaction chambers. When filling the reaction chambers, the form of a flow guiding column is adopted to break the fluid inertia and the surface tension between the fluid and the channel wall, realizing the complete filling of the reaction chambers from bottom to top and from the inlet end to the outlet end by the liquid without generating bubbles. After the sample distribution and filling are completed in each reaction chamber, there is a pressure from the inlet end to the outlet end, so that there will be no diversion and intermixing of samples during the subsequent amplification and reaction processes, avoiding cross-reactions.
[0023] 2. The microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acid is provided with a snap ring outside the upper shell. After placing the pressing plate, only by rotating the snap ring to clamp the snap bar can the pressing plate be firmly fixed at the bottom of the reaction base, and then keep multiple reaction cartridges tightly pressed at the bottom, making the installation, disassembly and cleaning of multiple reaction cartridges relatively convenient.
[0024] 3. The microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acid is further provided with a leakage absorption groove around the card seat. Even if there is liquid leakage between the sample tube and the hollow guiding column, it will be blocked in the leakage absorption groove and adsorbed by the sponge ring, avoiding the liquid in the leakage absorption groove from sloshing out during the movement. Description of the Drawings
[0025] Figure 1 is the assembly drawing of the present utility model;
[0026] Figure 2 is the exploded view of the present utility model;
[0027] Figure 3 is the cross-sectional view of the reaction base of the present utility model;
[0028] Figure 4 is the cross-sectional view of the sample tube of the present utility model;
[0029] Figure 5 is the schematic diagram of the flow splitting component of the present utility model.
[0030] In the figure: 1 - sample tube, 11 - sealing cap, 12 - valve;
[0031] 2 - reaction base, 21 - upper shell, 22 - bottom cover, 23 - card seat, 24 - overflow prevention hole, 25 - overflow prevention filter element, 26 - limiting strip, 27 - leakage absorption groove, 28 - sponge ring;
[0032] 3 - flow splitting component, 31 - hollow guiding column, 32 - fixing piece, 33 - flow guiding column, 34 - flow splitting tube, 35 - diversion branch tube;
[0033] 4 - pressing plate, 41 - snap bar;
[0034] 5 - Reaction cartridge;
[0035] 6 - Snap ring, 61 - Limit groove, 62 - Card slot. Specific embodiments
[0036] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.
[0037] Refer to Figures 1-5 , the present invention provides the following three technical solutions:
[0038] The first embodiment: A microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acids, comprising:
[0039] Sample tube 1, used to accommodate the sample of pathogenic microorganism nucleic acid to be detected or the reagent for sample treatment. For example, swab samples and sample preservation solutions, or liquid state samples, or DNA / RNA samples after nucleic acid extraction;
[0040] Reaction base 2, used to split and react the sample in sample tube 1;
[0041] The top of sample tube 1 is connected with a sealing cover 11 through a flexible connector on one side, and a valve 12 is arranged at the bottom of sample tube 1. Reaction base 2 is connected to the bottom end of sample tube 1 by plugging and pushes open valve 12 to make the liquid in sample tube 1 flow out. A flow splitting component 3 for uniformly splitting and reacting the liquid flowing out of sample tube 1 is arranged inside reaction base 2.
[0042] Reaction base 2 includes an upper shell 21 and a bottom cover 22, and the upper shell 21 and the bottom cover 22 are fixedly connected by bolts. A clamping seat 23 for clamping sample tube 1 is arranged in the middle of the top of upper shell 21.
[0043] A plurality of anti - overflow holes 24 are recessed in the outer periphery of the top of upper shell 21, and an anti - overflow filter element 25 that expands after absorbing liquid is plugged in anti - overflow holes 24.
[0044] The flow splitting component 3 includes:
[0045] The hollow guide post 31 is fixedly connected through the center of the upper shell 21 and is used to push open the valve 12. The top of the hollow guide post 31 is provided with an opening, and a sealing structure is arranged near the top on the side of the hollow guide post 31 to prevent liquid leakage. When inserting the sample tube 1 downward, the hollow guide post 31 will first push open the valve 12 and insert into the sample tube 1. As the pressure is applied downward, the liquid pressure in the sample tube 1 increases, promoting the liquid to enter the hollow guide post 31;
[0046] The fixing pieces 32 are provided in plurality and are evenly distributed at the bottom of the bottom cover 22;
[0047] The diversion column 33 is fixedly connected to one side of the bottom of the fixing piece 32;
[0048] The shunt tube 34 is connected to the hollow guide post 31 and the diversion column 33 at both ends respectively;
[0049] One end of the drainage branch tube 35 is fixedly connected through the other side of the diversion column 33, and the other end of the drainage branch tube 35 communicates with the bottom of the side of the anti-overflow hole 24.
[0050] A pressing plate 4 is detachably arranged at the bottom of the reaction base 2. A plurality of reaction boxes 5 are evenly penetrated in the circumferential direction inside the pressing plate 4. The reaction boxes 5 are made of a transparent thin-walled material, which is conducive to heat transfer and optical signal detection. In different reaction boxes 5, nucleic acid amplification reagents and different primer probes can be pre-placed to detect different targets. The pressing plate 4 presses the reaction boxes 5 against the bottom of the fixing piece 32 and realizes sealing. Nucleic acid amplification reagents and primer probes are pre-placed in the reaction boxes 5 before detection.
[0051] The liquid entering the hollow guide post 31 randomly enters one or more shunt tubes 34 and then enters the diversion column 33. Since the bottom end of the diversion column 33 extends to the bottom of the inner cavity of the reaction box 5, the fluid will further generate surface tension with the side wall of the reaction box 5, making the fluid close to the side wall of the reaction box 5 and vertically injecting into the reaction box 5. Thus, the subsequent further fluid will fill the entire reaction box 5 evenly and completely from bottom to top and from the inlet end to the outlet end without generating bubbles, and at the same time be fully mixed with the nucleic acid amplification reagents and primer probes pre-placed therein.
[0052] The fluid that has completed filling the reaction cartridge 5 further flows into the drainage branch pipe 35 and contacts the anti-overflow filter element 25 through the anti-overflow hole 24. When the anti-overflow filter element 25 encounters the fluid, the water-absorbing molecular structure inside it will change, causing its volume to rapidly expand and sealing the originally unobstructed reagent flow path, thereby completing the sealing of one or more drainage branch pipes 35. Further, the subsequent flowing fluid will choose the unfilled drainage branch pipes 35, and finally fill all the reaction cartridges 5 and all the drainage branch pipes 35. And because there is still pressure between the sample main injection port and the end of the anti-overflow filter element 25 of each drainage branch pipe 35 at this time, the reagents between each chamber will not flow back and mix, that is, no cross-contamination will occur. The microfluidic cartridge in this state can be used for subsequent amplification and detection.
[0053] This device only requires one-step sample addition and assembly actions to complete the injection of the sample and the equal distribution of multiple reaction chambers; when filling the reaction chambers, the form of the diversion column 33 is used to break the fluid inertia and the surface tension between the fluid and the channel wall, realizing the complete filling of the liquid from the bottom up and from the inlet end to the outlet end of the reaction chamber without generating bubbles; after the sample is distributed and filled in each reaction chamber, there is pressure from the inlet end to the outlet end, so that there will be no diversion and mixing of the sample during the subsequent amplification and reaction processes, avoiding cross-reactions.
[0054] The second implementation mode is mainly different from the first implementation mode in that: a snap ring 6 is rotatably sleeved on the outside of the upper shell 21, a limiting strip 26 penetrating the snap ring 6 is fixedly connected to the side surface of the upper shell 21, and a limiting groove 61 for restricting the rotation angle of the limiting strip 26 is provided inside the snap ring 6. A plurality of clamping strips 41 are uniformly arranged on the outside of the pressing plate 4, and a plurality of clamping grooves 62 for rotatably engaging the clamping strips 41 are uniformly provided on the snap ring 41 for connecting the pressing plate 4 to the bottom of the upper shell 21.
[0055] By arranging the snap ring 6 outside the upper shell 21, after placing the pressing plate 4, only need to rotate the snap ring 6 to clamp the clamping strip 41, then the pressing plate 4 can be firmly fixed at the bottom of the reaction base 2, and further keep the multiple reaction cartridges 5 tightly pressed at the bottom, making the installation, disassembly and cleaning of the multiple reaction cartridges 5 more convenient.
[0056] The third implementation mode is mainly different from the first implementation mode in that: a leakage absorption groove 27 is recessed at the top of the reaction base 2 and around the outer periphery of the card seat 23, and a sponge ring 28 is provided in the leakage absorption groove 27.
[0057] By further providing the leakage absorption groove 27 around the outer periphery of the card seat 23, even if liquid leakage occurs between the sample tube 1 and the hollow guide post 31, it will be blocked in the leakage absorption groove 27 and adsorbed by the sponge ring 28, avoiding the liquid in the leakage absorption groove 27 from sloshing out during the movement.
[0058] Meanwhile, the content not described in detail in this specification belongs to the prior art well-known to those skilled in the art.
[0059] It should be noted that, in this text, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not explicitly listed, or further includes elements inherent to such process, method, article or device.
[0060] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms, characterized in that, Comprising: A sample tube for accommodating a nucleic acid sample of a pathogenic microorganism to be detected or a reagent for sample treatment; A reaction base for shunting and reacting the sample in the sample tube; The top of the sample tube is connected with a sealing cap through a flexible connector on one side, and a valve is arranged at the bottom of the sample tube. The reaction base is connected to the bottom end of the sample tube in a plug-in manner and pushes open the valve to make the liquid in the sample tube flow out. A shunting component for uniformly shunting and reacting the liquid flowing out of the sample tube is arranged inside the reaction base.
2. The microfluidic cartridge for multiplex detection of nucleic acid of pathogenic microorganisms according to claim 1, characterized in that: The reaction base includes an upper shell and a bottom cover, and the upper shell and the bottom cover are fixedly connected by bolts. A clamping seat for clamping the sample tube is arranged in the middle of the top of the upper shell.
3. The microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acid according to claim 2, characterized in that: A plurality of anti-overflow holes are recessed in the outer periphery of the top of the upper shell, and anti-overflow filter elements that expand after absorbing liquid are plugged in the anti-overflow holes.
4. The microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms according to claim 1, characterized in that: The shunting component includes: A hollow guiding column fixedly connected through the center of the upper shell for pushing open the valve, and the top of the hollow guiding column is provided with an opening; Fixed pieces, with a plurality of them evenly distributed at the bottom of the bottom cover; A guiding column fixedly connected to one side of the bottom of the fixed piece; A shunting tube, with both ends respectively connected to the hollow guiding column and the guiding column; A drainage branch pipe, with one end fixedly connected through the other side of the guiding column, and the other end of the drainage branch pipe communicated with the bottom of the side of the anti-overflow hole.
5. The microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms according to claim 4, characterized in that: A pressing plate is detachably arranged at the bottom of the reaction base. A plurality of reaction boxes are evenly penetrated in the circumferential direction inside the pressing plate, and the pressing plate presses the reaction boxes against the bottom of the fixed piece to achieve sealing. Nucleic acid amplification reagents and primer probes are preset in the reaction boxes before detection.
6. The microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms according to claim 5, wherein: A clamping ring is rotatably sleeved on the outer side of the upper shell. A limiting strip penetrating through the clamping ring is fixedly connected to the side of the upper shell, and a limiting groove for restricting the rotation angle of the limiting strip is arranged inside the clamping ring.
7. The microfluidic cartridge for multiplex detection of pathogenic microorganism nucleic acid according to claim 6, wherein: A plurality of clamping strips are evenly arranged on the outer side of the pressing plate, and a plurality of clamping grooves for rotatably clamping the clamping strips are evenly arranged on the clamping ring for connecting the pressing plate to the bottom of the upper shell.
8. The microfluidic cartridge for multiplex detection of nucleic acids of pathogenic microorganisms according to claim 1, characterized in that: A leakage absorption groove is recessed at the top of the reaction base and around the clamping seat, and a sponge ring is arranged in the leakage absorption groove.