Integrated microfluidic detection device

Abstract

The utility model relates to an integrated micro-fluidic detection device, which belongs to the technical field of nucleic acid detection, and comprises a sample injection control device which comprises a sample injection control component, and at least four liquid storage cylinders and a power module which are connected to a sample injection control panel of the sample injection control component; the reaction control device is provided with a splitter plate attached to the sample injection control plate; the splitter plate is provided with sample injection runners corresponding to the bottoms of the liquid storage cylinders, the sample injection runners are communicated and converged to a column membrane chamber through a first micro valve, and a silicon-based adsorption membrane for adsorbing nucleic acid is arranged in the column membrane chamber; the lower-layer control plate is provided with a control flow channel communicated with the column membrane chamber, and the control flow channel is also respectively communicated with the PCR amplification chamber and the waste liquid chamber through a second micro valve; wherein the power module is arranged at the top position of the corresponding column film cavity.

Description

Technical Field

[0001] This utility model relates to an integrated microfluidic detection device, belonging to the field of nucleic acid detection technology. Background Technology

[0002] Nucleic acid testing is the gold standard for virus detection. However, the conventional processes of nucleic acid extraction, amplification, and detection require expensive equipment and are time-consuming and labor-intensive. In recent years, fully automated nucleic acid testing based on microfluidic test kits has attracted widespread attention. On the one hand, compared with traditional diagnostic technologies, fully automated microfluidic test kits for nucleic acid testing are characterized by high integration, small size, and portability, making them applicable to various scenarios where medical facilities are not yet fully developed. On the other hand, the entire testing process using microfluidic test kits is automated, improving the efficiency of nucleic acid testing and reducing manual operation costs.

[0003] Existing microfluidic detection reagent cartridges have low integration, are complex to use, and require manual processing beforehand, making the entire detection process time-consuming. Summary of the Invention

[0004] The purpose of this invention is to propose a highly integrated microfluidic detection reagent cartridge that integrates the entire nucleic acid extraction, amplification, and detection process to achieve fully automated operation.

[0005] An integrated microfluidic detection device shall have at least the following features:

[0006] The sample injection control device includes a sample injection control assembly and at least four liquid storage cylinders and a power module connected to the sample injection control plate of the sample injection control assembly; and

[0007] A reaction control device, comprising a flow divider plate arranged in conjunction with the sample injection control plate; the flow divider plate having sample injection channels corresponding to the bottom of each of the reservoirs, and several sample injection channels being connected and converged to a column membrane chamber via a first micro-valve, wherein a silicon-based adsorption membrane for adsorbing nucleic acids is provided in the column membrane chamber; and

[0008] The lower control board has a control channel that connects to the column membrane chamber, and the control channel is also connected to the PCR amplification chamber and the waste liquid chamber through a second microvalve, respectively.

[0009] Wherein: the power module is configured at the top of the corresponding diaphragm chamber.

[0010] The first microvalve and / or the second microvalve are respectively provided with elastic pressing control components and elastic pressing components, which are used to control the opening and closing of the first microvalve and the second microvalve. The power module can be a cylinder or a piston rod.

[0011] The elastic pressing control component and / or the elastic pressing component is a polymer film.

[0012] A filter mesh is installed between the silicon-based adsorption membrane and the control channel.

[0013] The front end of the PCR amplification chamber is a serpentine flow channel.

[0014] The elastic pressing component is disposed on the diverter plate or as a separate support plate.

[0015] The bottom of the injection channel is higher than the bottom of the column membrane chamber.

[0016] The liquid storage cylinder has an outer cylinder and an inner cylinder. The bottom of the inner cylinder is a thin film structure, and the bottom of the outer cylinder is equipped with a piercing device that can pierce the thin film structure at the bottom of the inner cylinder.

[0017] The outer cylinder and the inner cylinder of the liquid storage are also provided with matching protrusions or slots at the top.

[0018] Ventilation holes are provided on the sample injection control plate, the reaction control plate, and the lower control plate.

[0019] Compared with the prior art, the beneficial effects of this utility model are: 1. It can simultaneously extract nucleic acids from DNA and RNA pathogens; 2. The piston pusher simultaneously enables gas communication and provides liquid driving force; 3. This cartridge can be used with automated instruments to achieve high-throughput sample detection; 4. The liquid storage unit inside the reagent cartridge is pre-packaged with nucleic acid extraction reagents to achieve rapid detection and has an efficient sample processing and detection process. Attached Figure Description

[0020] Figure 1 An exploded view of the three-dimensional structure of an integrated microfluidic detection device;

[0021] Figure 2 This is a three-dimensional structural diagram of the sample injection control device 100;

[0022] Figure 3 This is a three-dimensional structural diagram of the reaction control device 200;

[0023] Figure 4 A three-dimensional structural diagram showing the connection between the sample injection control device 100 and the reaction control device 200;

[0024] Figure 5 This is a magnified view of the lower layer structure.

[0025] Figure 6 This is a magnified view of the lower layer structure.

[0026] Figure 7 A three-dimensional structural diagram of the integrated microfluidic detection device after installation;

[0027] Figure 8 for Figure 7 Enlarged view of section C in the embodiment;

[0028] Figure 9 A three-dimensional structural diagram of a liquid storage cylinder with a puncture device;

[0029] Figure 10 This is the overall structure diagram.

[0030] Wherein: 100-sample injection control device, 110-sample injection control component, 111-sample injection control board, 112-flow channel control port; 120-liquid storage cylinder, 121-outer cylinder, 1211-protruding structure, 122-liquid storage inner cylinder, 1221-slot, 130-power module;

[0031] Reaction control device, 210-diverter plate, 211-column membrane chamber, 212-first micro valve, 220-reaction control plate, 221-filter mesh, 222-reaction control port;

[0032] 300-Lower control board, 310-Control flow channel, 320-Waste liquid chamber, 330-Amplification chamber, 340-Second micro valve, 350-Elastic pressing component; 400-Elastic pressing control component, 600-Silicone film adsorption membrane, 700-Blocking ring, 800-Ventilation hole. Detailed Implementation

[0033] This invention is based on the principle of nucleic acid extraction using a column membrane method. The silica-based material used in this method adsorbs nucleic acids under high salt and low pH conditions, and releases them under low salt and high pH conditions. Under normal circumstances, the surface of nucleic acids is covered by a hydrophilic membrane composed of water molecules to maintain their water solubility. Under high salt concentrations and low pH (≤7.0), the nucleic acid effectively binds to the silica membrane due to the cation bridging effect formed by the salt ions (both the silica membrane and nucleic acid are negatively charged), while proteins, metabolites, and other contaminants cannot bind and are eluted. The basic process includes: cell membrane or cell wall lysis and nuclear envelope lysis; for viruses, after homogenization of the tissue, further lysis of the envelope and nucleocapsid is performed; removal of contaminants and stabilization of deoxyribonucleic acid (DNA); nucleic acid precipitation, washing, and elution.

[0034] like Figure 1 As shown, the integrated microfluidic detection device has at least the following features:

[0035] The sample injection control device 100 includes an injection control component 110 and at least four reservoirs 120 and a power module 130 connected to an injection control plate 111 of the injection control component 110. The four reservoirs are used to store lysis buffer, protein removal buffer, washing buffer, and elution buffer, respectively. The lysis buffer's main function is to disrupt cell structure and release intracellular nucleic acids. It typically contains components that can disrupt cell membranes and organelles, such as guanidine isothiocyanate, SDS (sodium dodecyl sulfate), and N-methylacetamide. These components effectively lyse cells, allowing for the full release of nucleic acids and thus improving the extraction efficiency and concentration. The protein removal buffer's main function is to remove proteins from the sample to reduce interference with nucleic acid extraction. It may contain components that denature or precipitate proteins, facilitating subsequent nucleic acid purification steps. The washing buffer is used for further cleaning and purification of nucleic acids. It removes residual proteins, cell debris, and other impurities through multiple washing steps, improving the purity of the nucleic acids. The primary function of eluent is to elute nucleic acids adsorbed onto a specific matrix for subsequent detection or analysis. Eluents typically possess properties that disrupt the interaction between nucleic acids and the adsorbent material, such as adjusting pH or ionic strength, thereby releasing the nucleic acids. The composition and concentration range of eluents can be 5-20 mM Tris-HCl, with a pH range of 6-8. For samples to be analyzed, they can be added to a reservoir 120 containing lysis buffer for lysis treatment.

[0036] It also includes a reaction control device 200, which has a flow divider 210 arranged in conjunction with the feed control plate 111; the flow divider 210 has a through-flowing membrane chamber 213 and a sample inlet channel 211 corresponding to the bottom of each of the liquid storage cylinders 120, and each sample inlet channel 211 is connected and converges to the membrane chamber 213; each sample inlet channel 211 is provided with a first micro valve 212, the open or closed state of which is used to control the flow of solution in the sample inlet channel 211. The upper end of the membrane chamber 213 is connected to a power module 130, which can be a cylinder or a piston rod, for applying air pressure to the membrane chamber 213, so that the solution therein is discharged through the silicon-based adsorption membrane 600 placed on one side of its bottom. Since each injection channel 211 is connected to four liquid storage cylinders 120, the corresponding solutions can be added sequentially by controlling the first micro valve 212 corresponding to the respective liquid storage cylinder 120, so that they enter the column membrane chamber 213.

[0037] In the column membrane chamber 213, a silicon-based adsorption membrane 600 is provided at the bottom. After the solution enters the column membrane chamber 213, nucleic acid can be adsorbed on the surface of the silicon-based adsorption membrane 600. Then, after passing through the protein removal solution and washing, the washing solution is discharged to the bottom. Then, the solution is eluted by the elution solution. The resulting desorbed solution containing nucleic acid is discharged downward to the PCR amplification chamber for amplification reaction.

[0038] The system also includes a lower control plate 300, which has a control channel 310 and a waste liquid chamber 320 connecting to the bottom side of the column membrane chamber 213, and at least one PCR amplification chamber 330. The control channel 310 can be connected to both the waste liquid chamber 320 and the PCR amplification chamber 330, and each connected channel is equipped with a second microvalve 340 for controlling the opening and closing of the channel, selectively draining the solution into the PCR amplification chamber 330 or into the waste liquid chamber 330. In at least one embodiment, the front end of the PCR amplification chamber 330 is a serpentine channel, and the PCR amplification chamber is used for nucleic acid amplification after receiving the elution buffer.

[0039] The power module 130 is positioned at the top of the corresponding diaphragm chamber 213, which allows it to... Figure 1 As can be seen, after the reagents in each of the storage cylinders 120 enter the membrane chamber 213, the pressure is applied by the pressing power module 130. Under pressure, the reagents in the membrane chamber 211 flow downward through the membrane chamber 213 and enter the control flow channel 310 after passing through the filter membrane in the membrane chamber 213. In one embodiment, the bottom of the membrane chamber 213 is also provided with a filter mesh for filtering unnecessary impurities or fine suspended matter in the solution.

[0040] In at least one embodiment, the reaction control device 200 further includes a reaction control plate 220 disposed between the flow divider plate 210 and the lower control plate 300. The reaction control plate 220 is provided with an elastic pressing control component 350 and a corresponding second microvalve 340 (e.g., for use in the control channel 310 of the lower control plate 300) within the control channel 310. Figure 5 This enables control over the working state of the control channel 310 within the lower control board 300.

[0041] In at least one embodiment, a first microvalve 212 is arranged within each of the sample inlet channels 211; the sample inlet control plate 111 is also provided with a channel control port 112 corresponding to each of the first microvalve 212, and each channel control port 112 is provided with an elastic pressing control component 400, which from Figure 1 and Figure 7As can be seen, in specific operation, the state of the first micro valve 212 is controlled by touching the elastic pressing control component 400 on the flow channel control port 112, thereby controlling the opening and closing state of each feed flow channel 212; the elastic pressing control component 400 here can be a PDMS membrane or other polymer membrane, and pressing can control the first micro valve 212 to open or close it.

[0042] In at least one embodiment, the control channel 310 is connected to the waste liquid chamber 320 and the amplification chamber 330 respectively through a second micro valve 340. The top of the second micro valve 340 is connected to an elastic pressing component 350. The elastic pressing component 350 can control the opening and closing of the micro valve by pressing, so as to directly control the working state of the second micro valve 340 in the control channel 310, the waste liquid chamber 320 and the amplification chamber 330.

[0043] The top of the diaphragm chamber 213 is connected to the power module 130 via a retaining ring 700, which... Figure 2 As can be seen, using the retaining ring 700 to elevate the power module 130 simplifies its installation structure, saves installation space, and makes the entire microfluidic detection device more concise. The retaining ring 700 is also used to fix the silicon membrane adsorption membrane 600.

[0044] The intermediate layer chip has an irregular structure, and the surface where the front flow channel is located is a 10mm high layer, which creates a height difference between the liquid and the filter membrane. In at least one embodiment, the bottom of the sample inlet channel 212 is higher than the bottom of the column membrane chamber 213, so that the liquid entering through the column membrane chamber 213 and the silicon membrane adsorption membrane 600 create a height difference, allowing the liquid to flow therein.

[0045] The liquid storage cylinder 120 has an outer cylinder 121 and an inner cylinder 122. The bottom of the inner cylinder 122 has a thin film structure. A piercing device is provided on the sample injection control plate 111 corresponding to the bottom of each liquid storage cylinder 120. Simultaneously, a piercing device capable of piercing the thin film structure at the bottom of the inner cylinder 122 is provided on the bottom of the outer cylinder 121. This is from... Figure 9 As can be seen, this allows it to quickly puncture the thin film structure at the bottom of the liquid storage inner cylinder 122.

[0046] In at least one embodiment, the outer cylinder 121 and the liquid storage inner cylinder 122 are also provided with matching protrusions 1211 or slots 1221 at their top positions to prevent relative movement. When in use, the bottom of the outer cylinder 121 can be configured with a thin film structure that can pierce the bottom of the liquid storage inner cylinder 122 by changing the limiting position.

[0047] In at least one embodiment, the elastic pressing control component 400 and / or the elastic pressing component 350 is a PDMS film.

[0048] In at least one embodiment, vent holes 800 are respectively provided on the injection control plate 111, the reaction control plate 220 and the lower control plate 300 to facilitate the venting of air during solvent flow and promote normal liquid flow.

[0049] During the manufacturing process of this microfluidic chip, the corresponding reagents are first loaded into the inner cylinders of four reservoirs 120 and then encapsulated. In use, the reservoir containing the lysis buffer is opened, and the collected sample solution is added to induce cell lysis. Then, the inner cylinder is pressed, causing the lower part to puncture the membrane, allowing the lysis buffer to drain into the lower sample inlet channel 211. Next, the first microvalve 212 on the sample inlet channel 211 corresponding to that reservoir is opened, allowing the lysed solution to enter the surface of the silicon-based adsorption membrane 600, where nucleic acids are adsorbed onto the membrane surface. Then, the power module is pressed, controlling the second microvalve. It is necessary to ensure communication between the control channel 310 and the waste liquid chamber 320, and simultaneously ensure communication between the control channel 310 and the PCR amplification chamber 33. The membrane between the 0 and 1 is closed, allowing the adsorbed solution to pass through the membrane and enter the waste liquid chamber. Then, the storage cylinders of the protein removal solution and washing solution are punctured in sequence. The power module is pressed in the same way to allow the used protein removal solution and washing solution to enter the waste liquid chamber 320. Finally, the bottom membrane of the storage cylinder containing the elution solution is punctured to elute the surface of the adsorption membrane, so that the adsorbed nucleic acid is eluted to obtain the desorbed solution. At this time, the second micro valve needs to be controlled to close the control channel 310 and the waste liquid chamber 320. At the same time, it is necessary to ensure the connection between the control channel 310 and the PCR amplification chamber 330, so that the desorbed solution enters the PCR amplification chamber. The amplified reaction solution can then be detected by fluorescence / qPCR detection method.

Claims

1. An integrated microfluidic detection device, characterized in that, At least has: Sample control device (100), the sample control device (100) includes sample control component (110) and at least four reservoirs (120) and power module (130) connected on the sample control board (111) of the sample control component (110);And Reaction control device (200), the reaction control device (200) has the shunt plate (210) arranged with the sample control board (111);The shunt plate (210) has the sample flow channel (211) configured at the bottom of each reservoir (120), and several sample flow channels (211) are communicated to the column membrane chamber (213) by the first micro valve (212), and the silicon-based adsorption membrane (600) for adsorbing nucleic acid is arranged in the column membrane chamber (213);And Lower control plate (300), the lower control plate (300) has control flow channel (310) communicated with the column membrane chamber (213), and the control flow channel (310) is also communicated with PCR amplification cavity (330) and waste cavity (320) by the second micro valve (340) respectively; Wherein: the power module (130) is arranged at the top position corresponding to the column membrane chamber (213).

2. The integrated microfluidic detection device of claim 1, wherein: The first micro valve (212) and / or the second micro valve (340) are respectively provided with elastic pressing control component (400) and elastic pressing component (350), for controlling the opening and closing of the first micro valve (212) and the second micro valve (340), and the power module (130) can be a gas cylinder or a piston push rod.

3. The integrated microfluidic detection device of claim 2, wherein: The elastic pressing control component (400) and / or the elastic pressing component (350) are polymer films.

4. The integrated microfluidic assay device of claim 1, wherein: The filter screen (221) is arranged between the silicon-based adsorption membrane (600) and the control flow channel (310).

5. The integrated microfluidic detection device of claim 3, wherein: The front end of the PCR amplification cavity (330) is a snake-shaped flow channel.

6. The integrated microfluidic assay device of claim 3, wherein: The elastic pressing component (350) is arranged on the shunt plate (210) or is a separate support plate.

7. The integrated microfluidic assay device according to any one of claims 1 to 6, wherein: The bottom of the sample flow channel (211) is higher than the bottom of the column membrane chamber (213).

8. The integrated microfluidic assay device of claim 7, wherein: The reservoir (120) has an outer cylinder (121) and a liquid storage inner cylinder (122), and the bottom of the liquid storage inner cylinder (122) is a film structure, and the bottom of the outer cylinder (121) is provided with a piercing device capable of piercing the film structure at the bottom of the liquid storage inner cylinder (122).

9. The integrated microfluidic assay device of claim 8, wherein: The top position of the outer cylinder (121) and the liquid storage inner cylinder (122) is also provided with a matching protruding structure (1211) or a clamping groove (1221).

10. The integrated microfluidic assay device of claim 1, wherein: Ventilation holes (800) are respectively formed in the sample control board (111), the reaction control plate (220) and the lower control plate (300).

Images