Universal Automated Nucleic Acid Extraction Module Based on Rotating Structure

Abstract

The present invention relates to the technical field of nucleic acid detection, and discloses a general-purpose automated nucleic acid extraction module based on a rotating structure, which includes a liquid storage layer and a flow channel layer arranged up and down; a lysis solution chamber, a binding solution chamber, a magnetic bead chamber, a pre-washing solution chamber, a post-washing solution chamber, an elution solution chamber, a sample processing chamber, a waste liquid chamber, and a loading chamber are provided in the nucleic acid extraction module; a valve core is installed in the sample processing chamber, a sample processing chamber piston is installed in the core cavity of the valve core, and a liquid transfer port communicating with the core cavity is opened on the side wall of the core cavity; flow channels communicating with the sample processing chamber are respectively opened on the top surface of the flow channel layer corresponding to the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, the elution solution chamber, the waste liquid chamber, and the loading chamber; the valve core rotates around its own axis and can communicate with different flow channels. The liquid flow channel switching is convenient, the structure is simpler, the assembly is simpler, the cost is lower, the volume is smaller, and the weight is lighter.

Description

Technical Field

[0001] The present invention relates to the technical field of nucleic acid detection, and particularly relates to a general-purpose automated nucleic acid extraction module based on a rotating structure. Background Art

[0002] Nucleic acid detection occupies an extremely important position in pathogen detection due to its high accuracy, sensitivity, and specificity, and it still exists as the gold standard for clinical diagnosis at present.

[0003] Nucleic acid extraction methods include cell lysis methods, centrifugal column methods, magnetic bead methods, etc. Among them, in the magnetic bead method, after surface modification and surface decoration of magnetic nanoparticles, they can reversibly bind and release nucleic acids under certain conditions. The magnetic bead method has the characteristics of simple operation, short time consumption, and can achieve automated and high-throughput operations, and the obtained nucleic acids have high purity and large concentration.

[0004] Manually dispensing reagents in the nucleic acid extraction process can no longer meet the market's demand for high throughput. Traditional nucleic acid extraction devices often require multiple valves (especially microvalves) to control liquid flow, and a highly integrated valve system may lead to a more complex overall structure of the device and an increase in cost. To accommodate numerous microvalves and their control circuits, the volume and weight of the device are often difficult to reduce, which limits its deployment and movement in a laboratory environment with limited space.

[0005] Therefore, there is an urgent need to improve traditional nucleic acid extraction devices to overcome the above defects. Summary of the Invention

[0006] The purpose of the present invention is to provide a general-purpose automated nucleic acid extraction module based on a rotating structure with simple liquid flow control, so as to reduce the setting of valves and reduce the volume and weight of the nucleic acid extraction module.

[0007] To solve the above technical problems, the technical solution of the present invention is as follows:

[0008] A general-purpose automated nucleic acid extraction module based on a rotating structure, the nucleic acid extraction module includes a liquid storage layer and a flow channel layer arranged in sequence from top to bottom;

[0009] A lysis solution chamber, a binding solution chamber, a magnetic bead chamber, a pre-washing solution chamber, a post-washing solution chamber, an elution solution chamber, a sample processing chamber, a waste liquid chamber, and a loading chamber are provided in the nucleic acid extraction module;

[0010] The sample processing chamber extends upward through the top surface of the liquid storage layer and extends downward into the flow channel layer. A valve core is installed in the sample processing chamber, a sample processing chamber piston is installed in the core cavity of the valve core, and a liquid transfer port communicating with the core cavity is opened on the side wall of the core cavity;

[0011] On the top surface of the flow channel layer, flow channels communicating with the sample processing chamber are respectively provided corresponding to the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, the elution solution chamber, the waste liquid chamber, and the loading chamber; wherein,

[0012] The valve core can rotate around its own axis to enable the liquid transfer port to communicate with different flow channels.

[0013] Further, a vortex-shaped flow channel is formed on the inner bottom wall of the core cavity of the valve core.

[0014] Further, the magnetic bead chamber is located between the binding solution chamber and the sample processing chamber, and the flow channel corresponding to the binding solution chamber extends through the flow channel corresponding to the magnetic bead chamber.

[0015] Further, the flow channels corresponding to the lysis solution chamber, the binding solution chamber, the pre-washing solution chamber, the post-washing solution chamber, and the elution solution chamber extend radially from one end of a main flow channel, and the other end of the main flow channel communicates with the sample processing chamber.

[0016] Further, the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, and the elution solution chamber respectively extend upward through the top surface of the liquid storage layer, and the nucleic acid extraction module further includes a flexible membrane layer provided on the bottom wall of the liquid storage layer, and the flexible membrane layer is used to seal the bottoms of the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, and the elution solution chamber located in the liquid storage layer.

[0017] Further, rubber stoppers are respectively installed in the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, and the elution solution chamber, and thorn-like structures are provided on their respective bottom walls, and the thorn-like structures are used to pierce the corresponding parts on the flexible membrane layer.

[0018] Further, a pressure buffer chamber is also provided in the nucleic acid extraction module, the pressure buffer chamber extends upward through the top surface of the liquid storage layer and downward through the bottom surface of the flow channel layer, a rubber stopper is provided in the pressure buffer chamber, and the rubber stopper can float up and down in the pressure buffer chamber.

[0019] Further, the nucleic acid extraction module further includes a gasket layer located between the flexible membrane layer and the liquid storage layer. Through holes are respectively formed in the gasket layer vertically corresponding to the pressure buffer chamber, the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, the elution solution chamber, the sample processing chamber, the waste liquid chamber, and the loading chamber.

[0020] Further, the waste liquid chamber extends upward through the top surface of the liquid storage layer and downward into the flow channel layer. A waste liquid chamber piston is installed in the waste liquid chamber.

[0021] Further, the loading chamber extends upward through the top surface of the liquid storage layer and downward through the bottom surface of the flow channel layer. A loading chamber piston is installed in the loading chamber.

[0022] Further, the nucleic acid extraction module further includes an electromagnet installed at the bottom of the sample processing chamber. The electromagnet can rotate around its own axis, and the valve core can rotate synchronously with the electromagnet.

[0023] The technical solution of the present invention has the following advantages:

[0024] 1. The general-purpose automated nucleic acid extraction module based on a rotating structure provided by the present invention. The sample processing chamber extends upward through the top surface of the liquid storage layer and downward into the flow channel layer. A valve core is installed in the sample processing chamber, and a sample processing chamber piston is installed in the core cavity of the valve core. A liquid transfer port communicating with the core cavity is formed on the side wall of the core cavity. Flow channels communicating with the sample processing chamber are respectively formed on the top surface of the flow channel layer corresponding to the lysis solution chamber, the binding solution chamber, the magnetic bead chamber, the pre-washing solution chamber, the post-washing solution chamber, the elution solution chamber, the waste liquid chamber, and the loading chamber. Among them, the valve core can rotate around its own axis to connect the liquid transfer port with different flow channels. In this way, the connection between the sample processing chamber and different chambers can be realized by the rotation of the valve core, and different liquid flow paths can be obtained. Compared with setting valves (especially micro-valves) for each chamber respectively, the structure is simpler, the assembly is simpler, the cost is lower, the volume is smaller, and the weight is lighter.

[0025] 2. The general-purpose automated nucleic acid extraction module based on a rotating structure provided by the present invention. A vortex-shaped flow channel is designed on the inner bottom wall of the valve cavity of the valve core, which can promote sufficient disturbance of the liquid flowing into the valve core to suspend the magnetic beads at the bottom of the valve core, thereby ensuring sufficient washing and effective elution of nucleic acids, and also helping to reduce non-specific binding, ensuring high purity of nucleic acid extraction.

[0026] 3. The universal automated nucleic acid extraction module based on a rotating structure provided by the present invention has a magnetic bead chamber located between a binding liquid chamber and a sample processing chamber. The flow channel corresponding to the binding liquid chamber extends through the flow channel corresponding to the magnetic bead chamber. In this way, when transferring liquid, the binding liquid will flow through the flow channel where the magnetic beads have flowed, so as to further rinse the flow channel corresponding to the magnetic bead chamber, thereby reducing the residue of magnetic beads in the flow channel corresponding to the magnetic bead chamber and improving the detection accuracy.

[0027] 4. The universal automated nucleic acid extraction module based on a rotating structure provided by the present invention is respectively provided with rubber stoppers in a lysis liquid chamber, a binding liquid chamber, a magnetic bead chamber, a pre-washing liquid chamber, a post-washing liquid chamber, and an elution liquid chamber, and a spiky structure is provided on the bottom wall of each of them. The spiky structure is used to pierce the corresponding part on the flexible membrane layer. The rubber stopper can not only push the flexible membrane layer towards the spiky structure so that the spiky structure pierces the flexible membrane layer, but also further squeeze out the liquid in each chamber, reducing the residue of the liquid in each chamber.

[0028] 5. The universal automated nucleic acid extraction module based on a rotating structure provided by the present invention further includes a gasket layer located between the flexible membrane layer and the liquid storage layer. Through holes are respectively opened on the gasket layer vertically corresponding to a pressure buffer chamber, a lysis liquid chamber, a binding liquid chamber, a magnetic bead chamber, a pre-washing liquid chamber, a post-washing liquid chamber, an elution liquid chamber, a sample processing chamber, a waste liquid chamber, and a loading chamber. The setting of this gasket provides more downward deformation space for the flexible membrane layer to ensure that the spiky structure can pierce the flexible membrane layer.

[0029] 6. The universal automated nucleic acid extraction module based on a rotating structure provided by the present invention has a waste liquid chamber extending upward through the top surface of the liquid storage layer and downward into the flow channel layer. A waste liquid chamber piston is installed in the waste liquid chamber. When waste liquid enters the waste liquid chamber, lifting the waste liquid chamber piston can accelerate the speed of the waste liquid flowing from the sample processing chamber into the waste liquid chamber, and at the same time buffer the pressure in the device.

[0030] 7. The universal automated nucleic acid extraction module based on a rotating structure provided by the present invention has a loading chamber extending upward through the top surface of the liquid storage layer and downward through the bottom surface of the flow channel layer. A loading chamber piston is installed in the loading chamber. When loading, the loading chamber piston can promote the liquid to move downstream and accelerate the loading. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for use in the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0032] Figure 1 Schematic perspective view of the three-dimensional combination of the nucleic acid providing module in the embodiment of the present invention;

[0033] Figure 2 Schematic exploded view of the three-dimensional nucleic acid providing module in the embodiment of the present invention;

[0034] Figure 3 Schematic perspective view of the three-dimensional nucleic acid providing module when both the rubber stopper and the piston expose the top surface of the liquid storage layer in the embodiment of the present invention;

[0035] Figure 4 Schematic perspective view of the valve core in the embodiment of the present invention;

[0036] Figure 5 Internal schematic view of the valve core in the embodiment of the present invention;

[0037] Figure 6 Top view schematic of the valve core in the embodiment of the present invention;

[0038] Figure 7 Schematic perspective view of the flow channel layer in the embodiment of the present invention.

[0039] Explanation of reference numerals:

[0040] 1. Liquid storage layer; 10. Pressure buffer chamber; 100. Buffer chamber rubber stopper; 11. Lysis solution chamber; 111. First rubber stopper; 12. Binding solution chamber; 121. Second rubber stopper; 13. Magnetic bead chamber; 131. Third rubber stopper; 14. Pre-washing solution chamber; 141. Fourth rubber stopper; 15. Post-washing solution chamber; 151. Fifth rubber stopper; 16. Elution solution chamber; 161. Sixth rubber stopper; 17. Sample processing chamber; 170. Valve core; 171. Sample processing chamber piston; 172. Sealing member; 173. Clamping post; 174. Liquid transfer port; 175. Vortex flow channel; 18. Waste liquid chamber; 181. Waste liquid chamber piston; 19. Sampling chamber; 191. Sampling chamber piston; 2. Flexible membrane layer; 3. Gasket layer; 4. Flow channel layer; 40. Main flow channel; 42. Electromagnet; 421. Card slot; 43. First flow channel; 44. Second flow channel; 45. Third flow channel; 46. Fourth flow channel; 47. Fifth flow channel; 48. Waste liquid flow channel; 49. Sampling flow channel; A. Spiky structure. Detailed implementation manners

[0041] Next, the technical solutions of the present invention will be clearly and completely described in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of 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.

[0042] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by terms such as "upper", "lower", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present invention 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 should not be construed as a limitation to the present invention. In addition, the terms "first", "second", "third", "fourth", "fifth", "sixth" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance.

[0043] In the description of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "installed", "connected", "coupled" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0044] In addition, the technical features involved in different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0045] As Figures 1 to 7 shown, the present invention provides a general-purpose automated nucleic acid extraction module based on a rotating structure (hereinafter referred to as the nucleic acid extraction module). The nucleic acid extraction module includes a liquid storage layer 1 and a flow channel layer 4. Inside the nucleic acid extraction module, there are a lysis solution chamber 11 for storing lysis solution (also used as a sample chamber), a binding solution chamber 12 for storing binding solution, a magnetic bead chamber 13 for storing magnetic beads, a pre-washing solution chamber 14 for storing washing solution, a post-washing solution chamber 15 for storing washing solution, an elution solution chamber 16 for storing elution solution, a sample processing chamber 17, a waste liquid chamber 18 for collecting the waste liquid generated during the reaction process, and a loading chamber 19 for collecting the final nucleic acid eluate. The sample processing chamber 17 extends upward through the top surface of the liquid storage layer 1 and downward through the bottom wall of the flow channel layer 4. A valve core 170 is installed inside the sample processing chamber 17. A sample processing chamber piston 171 is installed inside the core cavity of the valve core 170, and a liquid transfer port 174 communicating with the core cavity is opened on the side wall of the core cavity. On the top surface of the flow channel layer 4, flow channels communicating with the sample processing chamber 17 are respectively opened corresponding to the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, the elution solution chamber 16, the waste liquid chamber 18, and the loading chamber 19. Among them, the valve core 170 can rotate around its own axis to make the liquid transfer port 174 communicate with different flow channels.

[0046] In the present invention, the sample processing chamber 17 extends upward through the top surface of the liquid storage layer 1 and downward through the bottom wall of the flow channel layer 4. A valve core 170 is installed in the sample processing chamber 17. A sample processing chamber piston 171 is installed in the core cavity of the valve core 170. A liquid transfer port 174 communicating with the core cavity is formed on the side wall of the core cavity; corresponding to the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, the elution solution chamber 16, the waste liquid chamber 18, and the sample loading chamber 19 on the top surface of the flow channel layer 4, flow channels communicating with the sample processing chamber 17 are respectively formed (as Figure 7 shown). Among them, the valve core 170 can rotate around its own axis so that the liquid transfer port 174 communicates with different flow channels. In this way, the sample processing chamber 17 can be communicated with different chambers (such as the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, the elution solution chamber 16, the waste liquid chamber 18, and the sample loading chamber 19) by rotating the valve core 170, obtaining different liquid flow paths to complete nucleic acid extraction. Compared with respectively providing valves (especially micro-valves) for each chamber, the structure is simpler, the assembly is simpler, the cost is lower, the volume is smaller, and the weight is lighter.

[0047] Furthermore, a seal 172 is provided on the outer wall of the valve core 170 (as Figure 4 shown. The seal 172 is an O-ring and the number is two. Of course, the number can also be other values), which can ensure the sealing performance of the sample processing chamber 17 when the valve core 170 rotates. In this embodiment, the bottom end surface of the valve core 170 is a closed end surface.

[0048] Furthermore, as Figures 4 to 6 shown, a vortex flow channel 175 is designed on the inner bottom wall of the valve cavity of the valve core 170. In this way, the liquid flowing into the valve core 170 can be sufficiently disturbed to suspend the magnetic beads at the bottom inside the valve core 170, thereby ensuring sufficient washing and effective elution of nucleic acids, and also helping to reduce non-specific binding, ensuring high purity of nucleic acid extraction.

[0049] Furthermore, the height of the liquid transfer port 174 is the same as the height of the flow channel, and the bottom of the liquid transfer port 174 is basically flush with the bottom wall of the flow channel.

[0050] Furthermore, the cross-sectional shape of the flow channel can be semi-circular but is not limited to semi-circular.

[0051] Further, the magnetic bead chamber 13 is located between the binding solution chamber 12 and the sample processing chamber 17. The flow channel corresponding to the binding solution chamber 12 and the flow channel corresponding to the magnetic bead chamber 13 are arranged collinearly. In this way, when the liquid is transferred, the binding solution will flow through the flow channel where the magnetic beads have flowed, so as to further rinse the flow channel corresponding to the magnetic bead chamber 13, and thus the residue of the magnetic beads in the flow channel corresponding to the magnetic bead chamber 13 can be reduced.

[0052] Further, as Figure 7 shown, the flow channels corresponding to the lysis solution chamber 11, the binding solution chamber 12, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16 extend radially from one end of a main flow channel 40. The other end of the main flow channel 40 communicates with the sample processing chamber 17. For the convenience of understanding, the flow channels corresponding to the lysis solution chamber 11, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16 are respectively referred to as the first flow channel 43, the third flow channel 45, the fourth flow channel 46, and the fifth flow channel 47. In this embodiment, since the flow channels corresponding to the binding solution chamber 12 and the magnetic bead chamber 13 are arranged collinearly, they are collectively referred to as the second flow channel 44. The flow channel between the sample processing chamber 17 and the waste liquid chamber 18 is referred to as the waste liquid flow channel 48, and the flow channel between the sample processing chamber 17 and the loading chamber 19 is referred to as the loading flow channel 49.

[0053] Further, the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16 respectively extend upward through the top surface of the liquid storage layer 1. The nucleic acid extraction module further includes a flexible membrane layer 2 provided on the bottom wall of the liquid storage layer 1, and the flexible membrane layer 2 is used to seal the bottom ends of the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16 located in the liquid storage layer 1.

[0054] Further, rubber stoppers are respectively installed in the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16, and a spiky structure A is provided on the bottom wall of each of them. The spiky structure A is used to pierce the corresponding part of the flexible membrane layer 2. Specifically, the diameter of the rubber stopper is equal to the diameter of the corresponding chamber to improve the sealing performance during the detection process. In this embodiment, the rubber stopper is a silica gel stopper.

[0055] For the convenience of understanding, the rubber stoppers in the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, and the elution solution chamber 16 are respectively referred to as the first rubber stopper 111, the second rubber stopper 121, the third rubber stopper 131, the fourth rubber stopper 141, the fifth rubber stopper 151, and the sixth rubber stopper 161.

[0056] Further, a pressure buffer chamber 10 is also provided in the nucleic acid extraction module. The pressure buffer chamber 10 extends upward through the top surface of the liquid storage layer 1 and downward through the bottom surface of the flow channel layer 4. A rubber plug is provided in the pressure buffer chamber 10 (the rubber plug in the buffer chamber is called the buffer chamber rubber plug 100), and the buffer chamber rubber plug 100 can float up and down in the pressure buffer chamber 10. The pressure buffer chamber 10 is used to buffer the pressure in the downstream amplification device or detection device during the sample loading process.

[0057] Further, the nucleic acid extraction module further includes a gasket layer 3 located between the flexible membrane layer 2 and the liquid storage layer 1. Through holes are respectively provided on the gasket layer 3 vertically corresponding to the pressure buffer chamber 10, the lysis solution chamber 11, the binding solution chamber 12, the magnetic bead chamber 13, the pre-washing solution chamber 14, the post-washing solution chamber 15, the elution solution chamber 16, the sample processing chamber 17, the waste liquid chamber 18, and the sample loading chamber 19. In this embodiment, the setting of this gasket layer 3 provides more downward deformation space for the flexible membrane layer 2 to ensure that the spiky structure A can pierce the flexible membrane layer 2.

[0058] Further, the waste liquid chamber 18 extends upward through the top surface of the liquid storage layer 1 and downward into the flow channel layer 4. A waste liquid chamber piston 181 is installed in the waste liquid chamber 18. By lifting the waste liquid chamber piston 181, the speed of the waste liquid flowing from the sample processing chamber 17 into the waste liquid chamber 18 can be accelerated. By pulling up the waste liquid chamber piston 181, the waste liquid can be transferred into the waste liquid chamber 18. The waste liquid chamber 18 can also act as an air chamber to buffer the pressure inside the device during the nucleic acid extraction process.

[0059] Further, the material of the flexible membrane layer 2 includes but is not limited to TPU, polyethylene, polypropylene, etc. The bonding method between the flexible membrane layer 2 and the liquid storage layer 1 includes but is not limited to double-sided tape, hot melt, or ultrasonic welding.

[0060] Further, the sample loading chamber 19 extends upward through the top surface of the liquid storage layer 1 and downward through the bottom surface of the flow channel layer 4. A sample loading chamber piston 191 is installed in the sample loading chamber 19. During sample loading, the sample loading chamber piston 191 can promote the liquid to move downstream and accelerate the sample loading.

[0061] Further, the nucleic acid extraction module further includes an electromagnet 42 installed at the bottom of the sample processing chamber 17. The electromagnet 42 can rotate around its own axis, and the valve core 170 can rotate synchronously with the electromagnet 42. Specifically, a card slot 421 is provided at the top of the electromagnet 42, and a card post 173 is provided at the bottom of the valve core 170. The card post 173 is engaged with the card slot 421 so that the electromagnet 42 can drive the valve core 170 to rotate synchronously. The electromagnet 42 can also adsorb magnetic beads. The bottom end of the electromagnet 42 is connected to a rotating device, and the rotating device is used to drive the electromagnet 42 to rotate.

[0062] The specific operation process of the general-purpose automated nucleic acid extraction module provided by the present invention is introduced as follows:

[0063] Rotate the valve core 170 to connect the liquid transfer port 174 to the first flow channel 43. Pass a syringe through the first rubber stopper 111, add the sample into the lysis solution chamber 11, and press down the first rubber stopper 111 to the bottom of the lysis solution chamber 11, so that the spiky structure A pierces the flexible membrane layer 2. The mixed solution of the sample and the lysis solution enters the first flow channel 43. At the same time, extract the sample processing piston 171 in the sample processing chamber 17 upward to transfer the mixed solution to the sample processing chamber 17 for lysis reaction;

[0064] After the lysis reaction ends, rotate the electromagnet 42 to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the second flow channel 44. Press down the second rubber stopper 121 and the third rubber stopper 131 to the bottoms of the binding solution chamber 12 and the magnetic bead chamber 13 respectively. At the same time, extract the sample processing piston 171 to transfer the binding solution and the magnetic beads to the valve core 170 for binding reaction;

[0065] During the binding reaction, the electromagnet 42 outside the bottom of the valve core 170 adsorbs the magnetic beads to the inner bottom surface of the valve core 170;

[0066] Rotate the electromagnet 42 to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the waste liquid flow channel 48. Stop rotating the electromagnet 42, press down the sample processing piston 171, and at the same time lift the waste liquid piston 181 upward to transfer the waste liquid to the waste liquid chamber 18;

[0067] After the waste liquid transfer is completed, rotate the electromagnet 42 to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the third flow channel 45. Stop rotating the electromagnet 42;

[0068] Press down the fourth rubber stopper 141, and at the same time extract the sample processing piston 171 to transfer the cleaning solution in the pre-cleaning solution chamber 14 to the sample processing chamber 17 for cleaning. During the cleaning process, the electromagnet 42 no longer adsorbs the magnetic beads to the inner bottom surface of the valve core 170, so that the nucleic acid can be fully cleaned;

[0069] Rotate the electromagnet 42 to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the waste liquid flow channel 48. At the same time, start the electromagnet 42 to adsorb the magnetic beads to the inner bottom surface of the valve core 170, press down the sample processing piston 171, and at the same time lift the waste liquid piston 181 upward to transfer the waste liquid to the waste liquid chamber 18;

[0070] After the waste liquid transfer is completed, rotate the electromagnet 42 to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the fourth flow channel 46;

[0071] Stop rotating the electromagnet 42, press down the fifth rubber plug 151, and extract the sample processing chamber piston 171 at the same time, and transfer the cleaning liquid to the sample processing chamber 17 for cleaning;

[0072] During the cleaning process, the electromagnet 42 is disconnected and the magnetic beads are no longer adsorbed to the inner bottom surface of the valve core 170, so that the nucleic acid can be fully cleaned;

[0073] The electromagnet 42 is rotated to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the waste liquid flow channel 48, and the electromagnet 42 is activated to adsorb the magnetic beads to the inner bottom surface of the valve core 170;

[0074] Stop rotating the electromagnet 42, press down the sample processing chamber piston 171, and lift up the waste liquid chamber piston 181 to transfer the waste liquid to the waste liquid chamber 18;

[0075] After the waste liquid transfer is completed, the rotating electromagnet 42 drives the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the fifth flow channel 47;

[0076] Stop rotating the electromagnet 42 and disconnect the electromagnet 42, so that the magnetic beads are no longer adsorbed to the inner bottom surface of the valve core 170, so that the nucleic acid can be fully eluted;

[0077] Press the sixth rubber stopper 161 downwards and extract the sample processing chamber piston 171 at the same time, transferring the eluent to the sample processing chamber 17 for elution;

[0078] After the elution is completed, the electromagnet 42 adsorbs the magnetic beads to the inner bottom surface of the valve core 170; and the electromagnet 42 is rotated to drive the valve core 170 to rotate, so that the liquid transfer port 174 is connected to the sample loading channel 49;

[0079] Stop rotating the electromagnet 42, press down the sample processing chamber piston 171, and pull up the sample loading chamber piston 191 to transfer the nucleic acid eluate to the sample loading chamber 19;

[0080] The rotating electromagnet 42 drives the valve core 170 to rotate, so that the liquid transfer port 174 is offset from the sample loading channel 49, and the sample loading chamber piston 191 is pressed to complete the sample loading process in the downstream device.

[0081] In the process of assembling the nucleic acid extraction module, the rubber plug is first installed in the corresponding chamber, and the top of the rubber plug does not expose the top surface of the liquid storage layer 1, and then the nucleic acid extraction module is inverted so that the rubber plug blocks the bottom of the corresponding chamber, and then the liquid is injected into the chamber, and each chamber is sealed with the flexible film layer 2. Compared with first placing the flexible film layer 2 at the bottom of the liquid storage layer 1, then placing the liquid storage layer 1 upright, then injecting liquid into the chamber, and finally plugging the rubber plug, this assembly process can avoid the deformation of the flexible film layer 2 caused by the insertion of the rubber plug into the chamber.

[0082] The present invention provides a general-purpose automated nucleic acid extraction module based on a rotating structure, which utilizes a rotation mechanism to achieve precise transfer of liquids between different channels, thereby significantly improving the degree of automation and processing efficiency.

[0083] The present invention provides a general-purpose automated nucleic acid extraction module based on a rotating structure. Since a vortex-shaped flow channel 175 is designed on the inner bottom wall of the valve cavity of the valve core 170, in this way, it can cause the liquid flowing into the valve core 170 to generate a vortex effect and sufficient disturbance to suspend the magnetic beads at the bottom inside the valve core 170, reduce the deposition of the magnetic beads, make the distribution of the magnetic beads in the liquid more uniform, and further ensure that the nucleic acid can be fully washed and effectively eluted. It also helps to reduce non-specific binding and ensures high purity of nucleic acid extraction.

[0084] Obviously, the above-mentioned embodiments are only examples for clear illustration and not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all the implementation manners here. And the obvious changes or modifications derived therefrom still fall within the protection scope of the present invention.

Claims

1. A general-purpose automated nucleic acid extraction module based on a rotating structure, characterized in that The nucleic acid extraction module includes a liquid storage layer (1) and a flow channel layer (4) arranged in sequence from top to bottom; The nucleic acid extraction module is provided with a lysis solution chamber (11), a binding solution chamber (12), a magnetic bead chamber (13), a pre-washing solution chamber (14), a post-washing solution chamber (15), an elution solution chamber (16), a sample processing chamber (17), a waste liquid chamber (18), and a sample loading chamber (19); The sample processing chamber (17) extends upward through the top surface of the liquid storage layer (1) and downward through the bottom wall of the flow channel layer (4). A valve core (170) is installed in the sample processing chamber (17). A sample processing chamber piston (171) is installed in the core cavity of the valve core (170), and a liquid transfer port (174) communicating with the core cavity is opened on the side wall of the core cavity; On the top surface of the flow channel layer (4), flow channels communicating with the sample processing chamber (17) are respectively opened corresponding to the lysis solution chamber (11), the binding solution chamber (12), the magnetic bead chamber (13), the pre-washing solution chamber (14), the post-washing solution chamber (15), the elution solution chamber (16), the waste liquid chamber (18), and the sample loading chamber (19); among them, The valve core (170) can rotate around its own axis to enable the liquid transfer port (174) to communicate with different flow channels; A vortex flow channel (175) is formed on the inner bottom wall of the core cavity of the valve core (170); The magnetic bead chamber (13) is located between the binding solution chamber (12) and the sample processing chamber (17), and the flow channel corresponding to the binding solution chamber (12) extends through the flow channel corresponding to the magnetic bead chamber (13).

2. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 1, wherein The flow channels corresponding to the lysis solution chamber (11), the binding solution chamber (12), the pre-washing solution chamber (14), the post-washing solution chamber (15), and the elution solution chamber (16) extend radially from one end of a main flow channel (40), and the other end of the main flow channel (40) communicates with the sample processing chamber (17).

3. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 1 or 2, characterized in that The lysis solution chamber (11), the binding solution chamber (12), the magnetic bead chamber (13), the pre-washing solution chamber (14), the post-washing solution chamber (15), and the elution solution chamber (16) respectively extend upward through the top surface of the liquid storage layer (1). The nucleic acid extraction module further includes a flexible membrane layer (2) provided on the bottom wall of the liquid storage layer (1), and the flexible membrane layer (2) is used to seal the bottoms of the lysis solution chamber (11), the binding solution chamber (12), the magnetic bead chamber (13), the pre-washing solution chamber (14), the post-washing solution chamber (15), and the elution solution chamber (16) located in the liquid storage layer (1).

4. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 3, wherein The lysis solution chamber (11), the binding solution chamber (12), the magnetic bead chamber (13), the pre-washing solution chamber (14), the post-washing solution chamber (15), and the elution solution chamber (16) are each provided with a rubber stopper, and a spiky structure (A) is provided on the bottom wall of each of them. The spiky structure (A) is used to pierce the corresponding part of the flexible membrane layer (2).

5. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 4, wherein A pressure buffer chamber (10) is further formed in the nucleic acid extraction module. The pressure buffer chamber (10) extends upward through the top surface of the liquid storage layer (1) and downward through the bottom surface of the flow channel layer (4). A rubber stopper is provided in the pressure buffer chamber (10), and the rubber stopper can float up and down in the pressure buffer chamber (10).

6. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 5, characterized in that The nucleic acid extraction module further includes a gasket layer (3) located between the flexible membrane layer (2) and the flow channel layer (4). Through holes are respectively formed in the gasket layer (3) vertically corresponding to the pressure buffer chamber (10), the lysis solution chamber (11), the binding solution chamber (12), the magnetic bead chamber (13), the pre-washing solution chamber (14), the post-washing solution chamber (15), the elution solution chamber (16), the sample processing chamber (17), the waste liquid chamber (18), and the sample loading chamber (19).

7. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 6, wherein, The waste liquid chamber (18) extends upward through the top surface of the liquid storage layer (1) and downward into the flow channel layer (4). A waste liquid chamber piston (181) is provided in the waste liquid chamber (18).

8. The general-purpose automated nucleic acid extraction module based on a rotating structure according to claim 6, wherein The sample loading chamber (19) extends upward through the top surface of the liquid storage layer (1) and downward through the bottom surface of the flow channel layer (4). A sample loading chamber piston (191) is provided in the sample loading chamber (19).

9. The general-purpose automated nucleic acid extraction module based on a rotating structure according to any one of claims 1, 2, or 4-8, characterized in that The nucleic acid extraction module further includes an electromagnet (42) installed at the bottom of the sample processing chamber (17). The electromagnet (42) can rotate around its own axis, and the valve core (170) can rotate synchronously with the electromagnet (42).

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