A nucleic acid extraction device

Abstract

The present invention relates to the technical field of nucleic acid extraction, and provides a nucleic acid extraction device, which at least includes: a reagent storage unit, having a plurality of sealed reagent carrying chambers for carrying different reagents; an extraction operation unit, having a connected sample elution chamber and a reaction chamber; the sample elution chamber having a sample loading port connected to the external environment; a sealing patch, which is arranged at the sample loading port, and the sealing patch is opened when the sample is loaded, and the sealing patch re-seals the sample loading port after the sample is loaded. In the nucleic acid extraction device provided by the present invention, the sealing patch is opened when the sample is loaded, and the sealing patch re-seals the sample loading port after the sample is loaded, which can prevent the pathogen nucleic acid in the nucleic acid extraction device from entering the air and avoid polluting the environment. Moreover, different reagents are independently stored in the sealed reagent carrying chambers, and the sealing of the nucleic acid extraction device will not be destroyed during the reagent addition process, which can prevent nucleases in the environment from entering the nucleic acid extraction device to degrade the pathogen nucleic acid.

Description

Technical Field

[0001] The invention relates to the technical field of nucleic acid extraction, and in particular to a nucleic acid extraction device. Background Art

[0002] Nucleic acid detection technology has the advantages of high sensitivity, strong specificity and high accuracy. It can realize the identification and analysis of species / genus / type, fragment / sequence / site with different resolutions according to the optimization of detection methods, screening of target genes and design of primer probes. Therefore, it is widely used in clinical testing, disease control and emergency response, animal husbandry and veterinary medicine and other fields. Nucleic acid detection technology specifically includes two parts: nucleic acid extraction and nucleic acid analysis. Among them, nucleic acid analysis technology has been rapidly innovating and developing in the past decade. According to different analysis objects, it includes sequence determination method, gene analysis method, etc.; according to different analysis methods, it includes optical signal determination method, electrochemical signal determination method, naked eye recognition method, etc.; according to different temperature control during the analysis process, it includes variable temperature method, isothermal method, room temperature method, etc. Nucleic acid extraction technology, as an important step in nucleic acid extraction, purification and enrichment, is the core support for the sensitivity, accuracy and specificity of nucleic acid analysis technology. In the prior art, EP tubes are required for nucleic acid extraction. However, due to the large number of reagent types and complicated operating steps, the EP tubes are opened many times during the extraction process. Nucleases in the environment can easily enter the nucleic acid extraction system and degrade the pathogen nucleic acid. At the same time, the pathogen nucleic acid in the EP tube can also easily enter the air, causing environmental pollution. Summary of the invention

[0003] Therefore, the technical problem to be solved by the present invention is that in the prior art, EP tubes are needed for nucleic acid extraction, but due to the large number of reagent types and complicated operating steps, the EP tubes are opened many times during the extraction process, and nucleases in the environment can easily enter the nucleic acid extraction system and degrade the pathogen nucleic acid; at the same time, the pathogen nucleic acid in the EP tube can also easily enter the air and pollute the environment, thereby providing a nucleic acid extraction device.

[0004] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0005] The present invention provides a nucleic acid extraction device, which at least comprises: a reagent storage unit, which has a plurality of sealed reagent carrying chambers for carrying different reagents; an extraction operation unit, which has a sample elution chamber and a reaction chamber connected to each other, wherein a part of the reagent carrying chamber is connected to the sample elution chamber, and another part of the reagent carrying chamber is connected to the reaction chamber; the sample elution chamber has a sample loading port connected to the external environment; a sealing patch, which is arranged at the sample loading port, the sealing patch is opened when loading the sample, and the sealing patch re-seals the sample loading port after loading the sample.

[0006] Furthermore, the reagent storage unit includes a stacked storage substrate and an aluminum foil; a plurality of blisters convex outwardly in a direction away from the aluminum foil are arranged on the storage substrate, and the space between the blisters and the aluminum foil is used to form the reagent carrying chamber.

[0007] Furthermore, the extraction operation unit includes a first fluid layer and a second fluid layer which are stacked, and the first fluid layer is arranged in contact with the aluminum foil; a plurality of reagent inlets corresponding one to one with the reagent carrying chambers are arranged on the first fluid layer; the sample elution chamber and the reaction chamber are arranged in the second fluid layer; a part of the reagent inlet is connected to the sample elution chamber, and another part of the reagent inlet is connected to the reaction chamber.

[0008] Furthermore, the extraction operation unit also includes a waste liquid chamber, which is arranged in the second fluid layer, and the waste liquid chamber is connected to the reaction chamber.

[0009] Furthermore, the nucleic acid extraction device also includes a control layer, which is arranged on the side of the second fluid layer facing away from the first fluid layer; a driving chamber is arranged in the control layer, and the driving chamber is communicated with the waste liquid chamber; a piston is arranged in the driving chamber, and when the piston moves in the driving chamber, a negative pressure is formed in the driving chamber to drive the reagent in the reagent carrying chamber to flow into the sample elution chamber or the reaction chamber, the liquid in the sample elution chamber to flow into the reaction chamber, and the liquid in the reaction chamber to flow into the waste liquid chamber.

[0010] Furthermore, the control layer is also provided with a connection interface for connecting the nucleic acid extraction device with the nucleic acid analysis device; a gas inlet and a nucleic acid outlet are provided in the connection interface; and the gas inlet and the nucleic acid outlet are both connected to the reaction chamber.

[0011] Furthermore, microvalves are provided between the sample elution chamber and the reaction chamber, between the reaction chamber and the nucleic acid outlet, between the reaction chamber and the gas inlet, and between the reaction chamber and the waste liquid chamber.

[0012] Furthermore, the nucleic acid extraction device also includes a shell, which includes an upper cover and a lower cover connected to each other, and the space between the upper cover and the lower cover is used to accommodate the reagent storage unit, the extraction operation unit and the control layer; the sample addition port is connected to the external environment through the upper cover; the upper cover is provided with a first control channel leading to the bubble, and different bubbles are squeezed through different first control channels to make the reagent in the bubble flow out.

[0013] Furthermore, a first flow hole and a second flow hole for connecting the upstream and downstream of the microvalve are arranged in the valve cavity of the microvalve; the valve cavity of the microvalve is an open structure, and an elastic membrane is arranged at the open part of the valve cavity, and the first flow hole and the second flow hole are blocked when the elastic membrane is squeezed downward; a second control channel leading to the valve cavity is arranged on the upper cover, and different elastic membranes are squeezed through different second control channels to control different microvalves to open or close.

[0014] Furthermore, the nucleic acid extraction device also includes a sticker, which is arranged on the surface of the upper cover and is used to encapsulate the first control channel and the second control channel on the upper cover.

[0015] The technical solution of the present invention has the following advantages:

[0016] In the nucleic acid extraction device provided by the present invention, a sealing patch is provided at the sample loading port of the sample elution chamber. The sealing patch is opened when loading the sample, and the sealing patch reseals the sample loading port after loading the sample, which can prevent the pathogen nucleic acid in the nucleic acid extraction device from entering the air and avoid polluting the environment. Moreover, different reagents are independently stored in a sealed reagent-carrying chamber, and the reagent-carrying chamber is directly connected to the sample elution chamber or the reaction chamber. The sealing of the nucleic acid extraction device will not be destroyed during the reagent addition process, which can prevent nucleases in the environment from entering the nucleic acid extraction device to degrade the pathogen nucleic acid, which is conducive to ensuring the quality of nucleic acid extraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0018] Figure 1 Schematic diagram of a nucleic acid extraction device (sealing sticker opened) in an embodiment of the present invention;

[0019] Figure 2 is an explosion diagram of a nucleic acid extraction device in an embodiment of the present invention;

[0020] Figure 3 is a schematic diagram of a driving chamber in a nucleic acid extraction device in an embodiment of the present invention;

[0021] Figure 4 is a schematic diagram of a reagent carrying chamber in a nucleic acid extraction device in an embodiment of the present invention;

[0022] Figure 5is a schematic diagram of a connection interface in a nucleic acid extraction device in an embodiment of the present invention;

[0023] Figure 6 Schematic diagram of a reagent inlet and a microvalve in a nucleic acid extraction device in an embodiment of the present invention;

[0024] Figure 7 is a schematic diagram of a flow channel in a nucleic acid extraction device in an embodiment of the present invention;

[0025] Figure 8 Schematic diagram of the first microvalve (open state) in the nucleic acid extraction device in an embodiment of the present invention;

[0026] Fig. 9 Schematic diagram of the first microvalve (closed state) in the nucleic acid extraction device in an embodiment of the present invention.

[0027] 1. Shell;

[0028] 11. Sealing sticker; 12. Sticker; 13. Upper cover; 14. Lower cover; 15. First control channel; 16. Second control channel;

[0029] 2. Reagent storage unit;

[0030] 21. Blister; 22. Aluminum foil;

[0031] 211a, first reagent carrying chamber; 211b, second reagent carrying chamber; 211c, third reagent carrying chamber; 211d, fourth reagent carrying chamber; 211e, fifth reagent carrying chamber; 211f, sixth reagent carrying chamber;

[0032] 3. Extraction operation unit;

[0033] 31. elastic membrane; 32. first fluid layer; 33. second fluid layer; 34. sample loading port; 35. sample elution chamber; 36. reaction chamber; 37. waste liquid chamber; 38. connection interface; 39. nucleic acid outlet; 40. gas inlet;

[0034] 321a, first flow channel; 321b, second flow channel; 321c, third flow channel; 321d, fourth flow channel; 321e, fifth flow channel; 321f, sixth flow channel; 321g, seventh flow channel; 321h, eighth flow channel; 321i, ninth flow channel; 321j, tenth flow channel; 321k, eleventh flow channel; 331a, twelfth flow channel; 331b, thirteenth flow channel; 331c, fourteenth flow channel; 341, fifteenth flow channel;

[0035] 322a, first microvalve; 322b, second microvalve; 322c, third microvalve; 322d, fourth microvalve;

[0036] 323a, first reagent inlet; 323b, second reagent inlet; 323c, third reagent inlet; 323d, fourth reagent inlet; 323e, fifth reagent inlet; 323f, sixth reagent inlet;

[0037] 4. Control layer; 41. Piston; 42. Drive chamber; 43. Gear;

[0038] 51. Valve chamber; 52. First flow hole; 53. Second flow hole. DETAILED DESCRIPTION

[0039] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are 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 cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0041] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" 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 a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0042] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0043] like Figures 1 to 9As shown, this embodiment provides a nucleic acid extraction device, which at least includes: a reagent storage unit 2, having a plurality of sealed reagent carrying chambers for carrying different reagents; an extraction operation unit 3, having a sample elution chamber 35 and a reaction chamber 36 connected to each other, wherein a part of the reagent carrying chamber is connected to the sample elution chamber, and another part of the reagent carrying chamber is connected to the reaction chamber 36; the sample elution chamber 35 has a sample loading port 34 connected to the external environment, for example, the sample loading port 34 can be arranged at the top of the sample elution chamber 35; a sealing sticker 11, which is arranged at the sample loading port 34, and the sealing sticker 11 is opened when the sample is loaded, and the sealing sticker 11 re-seals the sample loading port 34 after the sample is loaded. Among them, the swab or the swab treatment liquid is added to the sample elution chamber 35 through the sample loading port 34. Among them, when in use, the dried binding reagent and the freeze-dried magnetic beads or enrichment membrane for capturing nucleic acids can be added to the reaction chamber 36 in advance.

[0044] In the nucleic acid extraction device provided in this embodiment, a sealing sticker 11 is provided at the sample loading port 34 of the sample elution chamber 35. The sealing sticker 11 is opened when loading the sample, and the sealing sticker 11 reseals the sample loading port 34 after loading the sample, which can prevent the pathogen nucleic acid in the nucleic acid extraction device from entering the air and avoid polluting the environment. In addition, different reagents are independently stored in a sealed reagent-carrying chamber, and the reagent-carrying chamber is directly connected to the sample elution chamber or the reaction chamber 36. The sealing of the nucleic acid extraction device will not be destroyed during the reagent addition process, which can prevent nucleases in the environment from entering the nucleic acid extraction device to degrade the pathogen nucleic acid, which is conducive to ensuring the quality of nucleic acid extraction.

[0045] like Figure 2 As shown, specifically, the reagent storage unit 2 includes a storage substrate and an aluminum foil 22 arranged in a stacked manner; a plurality of blisters 21 convex in a direction away from the aluminum foil 22 are arranged on the storage substrate, and the space between the blisters 21 and the aluminum foil 22 is used to form a reagent carrying chamber. For example, the number of reagent carrying chambers can be selected, for example, the number of reagent carrying chambers can be 5, or 6, etc. Wherein, the volume of each reagent carrying chamber can be 50μl or more, and the volumes of different reagent carrying chambers can be the same or different. Wherein, the shape of the reagent carrying chamber can be a cylinder, an elliptical cylinder or an irregular shape, and the cross-sections of different reagent carrying chambers can be the same or different. Wherein, the reagent carried in the reagent carrying chamber can be a liquid or a dry powder, and the types and types of reagents in different reagent carrying chambers can be the same or different. Wherein, the storage substrate and the aluminum foil 22 can be fixed together by thermal packaging technology to independently store and effectively seal the reagents in the reagent carrying chamber. The reagent storage unit 2 can be fixed on the top of the extraction operation unit 3 by means of snapping or bonding. When the reagent carrying chamber is pressed by external force, the bottom aluminum foil 22 can be broken, so that the liquid reagent can be used quickly.

[0046] like Figure 6 As shown, the extraction operation unit 3 includes a first fluid layer 32 and a second fluid layer 33 which are stacked, and the first fluid layer 32 is arranged in contact with the aluminum foil 22; the first fluid layer 32 is provided with a plurality of reagent inlets corresponding to the reagent carrying chambers; the sample elution chamber 35 and the reaction chamber 36 are arranged in the second fluid layer 33; a part of the reagent inlets is connected to the sample elution chamber 35, and another part of the reagent inlets is connected to the reaction chamber 36. When the bottom aluminum foil 22 of the reagent carrying chamber is crushed, the reagent can enter the sample elution chamber 35 or the reaction chamber 36 through the reagent inlet.

[0047] The extraction operation unit 3 further includes a waste liquid chamber 37, which is disposed in the second fluid layer 33, and the waste liquid chamber 37 is connected to the reaction chamber 36. For example, a porous water-absorbing material can be disposed in the waste liquid chamber 37 to absorb the liquid flowing through the chamber.

[0048] like Figure 3 As shown, the nucleic acid extraction device further includes a control layer 4, which is arranged on the side of the second fluid layer 33 facing away from the first fluid layer 32; a driving chamber 42 is arranged in the control layer 4, and the driving chamber 42 is connected to the waste liquid chamber 37; a piston 41 is arranged in the driving chamber 42, and when the piston 41 moves in the driving chamber 42, it is used to form a negative pressure in the driving chamber 42, so as to drive the reagent in the reagent carrying chamber to flow into the sample elution chamber 35 or the reaction chamber 36, the liquid in the sample elution chamber 35 to flow into the reaction chamber 36, and the liquid in the reaction chamber 36 to flow into the waste liquid chamber 37. For example, the piston 41 can be driven to move by a cylinder or a gear 43, a rack and a motor, and when the piston 41 moves outward, it can drive the liquid at the target position to flow.

[0049] like Figure 3 , Figure 5 As shown, the control layer 4 is also provided with a connection interface 38 for connecting the nucleic acid extraction device with the nucleic acid analysis device; a gas inlet 40 and a nucleic acid outlet 39 are provided in the connection interface 38; the gas inlet 40 and the nucleic acid outlet 39 are both connected to the reaction chamber 36. The nucleic acid outlet 39 is used to discharge the extracted nucleic acid, and the gas inlet 40 is used to evacuate the gas in the nucleic acid analysis device. For example, the connection interface 38 can be a screw or a socket, which facilitates the connection while ensuring the sealing of the connection.

[0050] Microvalves are disposed between the sample elution chamber 35 and the reaction chamber 36 , between the reaction chamber 36 and the nucleic acid outlet 39 , between the reaction chamber 36 and the gas inlet 40 , and between the reaction chamber 36 and the waste liquid chamber 37 .

[0051] like Figure 1 , Figure 2 as well as Figure 6 As shown, the nucleic acid extraction device also includes a shell 1, which includes an upper cover 13 and a lower cover 14 connected to each other, and the space between the upper cover 13 and the lower cover 14 is used to accommodate a reagent storage unit 2, an extraction operation unit 3 and a control layer 4; the sample injection port 34 is connected to the external environment through the upper cover 13; the upper cover 13 is provided with a first control channel 15 leading to the bubble 21, and different bubbles 21 are squeezed through different first control channels 15 to make the reagent in the bubble 21 flow out.

[0052] like Figure 1 , Figure 8 as well as Fig. 9 As shown, a first flow hole 52 and a second flow hole 53 for connecting the upstream and downstream of the microvalve are arranged in the valve cavity 51 of the microvalve; the valve cavity 51 of the microvalve is an open structure, and an elastic membrane 31 is arranged at the open part of the valve cavity 51. When the elastic membrane 31 is pressed downward, the first flow hole 52 and the second flow hole 53 are blocked; a second control channel 16 leading to the valve cavity 51 is arranged on the upper cover 13, and different elastic membranes 31 are squeezed through different second control channels 16 to control different microvalves to open or close. For example, the elastic membrane 31 can be a whole membrane, which is arranged between the aluminum foil 22 and the first fluid layer 32. On the one hand, it can be used for controlling the microvalve, and on the other hand, it can also improve the connection sealing between the aluminum foil 22 and the first fluid layer 32.

[0053] The nucleic acid extraction device further includes a sticker 12, which is arranged on the surface of the upper cover 13 and is used to encapsulate the first control channel 15 and the second control channel 16 on the upper cover 13. Such an arrangement can not only realize the encapsulation of the top of the upper cover 13, but also serve as a way to identify whether the nucleic acid extraction device is effective.

[0054] like Figure 4 , Figure 6 as well as Figure 7As shown, it should be noted that the nucleic acid extraction device in the present application also has a number of flow channels for connecting different chambers, which are either arranged in the first fluid layer 32, or arranged in the second fluid layer 33, or arranged in the control layer 4. The nucleic acid extraction device has 6 reagent carrying chambers as an example for explanation. The 6 reagent carrying chambers are respectively recorded as the first reagent carrying chamber 211a, the second reagent carrying chamber 211b, the third reagent carrying chamber 211c, the fourth reagent carrying chamber 211d, the fifth reagent carrying chamber 211e and the sixth reagent carrying chamber 211f. The corresponding first fluid layer 32 has 6 reagent inlets, which are respectively recorded as the first reagent inlet 323a, the second reagent inlet 323b, the third reagent inlet 323c, the fourth reagent inlet 323d, the fifth reagent inlet 323e and the sixth reagent inlet 323f. Among them, 4 microvalves are set, which are respectively recorded as the first microvalve 322a, the second microvalve 322b, the third microvalve 322c and the fourth microvalve 322d. Among them, 15 flow channels are provided, which are respectively recorded as the first flow channel 321a, the second flow channel 321b, the third flow channel 321c, the fourth flow channel 321d, the fifth flow channel 321e, the sixth flow channel 321f, the seventh flow channel 321g, the eighth flow channel 321h, the ninth flow channel 321i, the tenth flow channel 321j, the eleventh flow channel 321k, the twelfth flow channel 331a, the thirteenth flow channel 331b, the fourteenth flow channel 331c and the fifteenth flow channel 341. Among them, the first flow channel 321a is located between the first reagent inlet 323a and the sample elution chamber 35. The second flow channel 321b is located between the second reagent inlet 323b and the sample elution chamber 35. The third flow channel 321c is located between the third reagent inlet 323c and the first microvalve 322a. The fourth flow channel 321d is located between the third reagent inlet 323c and the reaction chamber 36. The fifth flow channel 321e is located between the fourth reagent inlet 323d and the reaction chamber 36. The sixth flow channel 321f is located between the fifth reagent inlet 323e and the reaction chamber 36. The seventh flow channel 321g is located between the sixth reagent inlet 323f and the reaction chamber 36. The eighth flow channel 321h is located between the fourth microvalve 322d and the waste liquid chamber 37. The ninth flow channel 321i is located between the third microvalve 322c and the waste liquid chamber 37. The tenth flow channel 321j is located between the third microvalve 322c and the inlet. The eleventh flow channel 321k is located between the second microvalve 322b and the outlet. The twelfth flow channel 331a is located between the sample elution chamber 35 and the first microvalve 322a. The thirteenth flow channel 331b is located between the reaction chamber 36 and the second microvalve 322b. The fourteenth flow channel 331c is located between the second microvalve 322b and the fourth microvalve 322d. The fifteenth flow channel 341 is located between the waste liquid chamber 37 and the driving chamber 42 .

[0055] Next, we will introduce the use of two different nucleic acid extraction reagents:

[0056] Embodiment 1

[0057] The six reagent-carrying chambers are all circular in shape, with diameters of 10 mm, 8 mm, 6 mm, 13 mm, 11 mm, and 12 mm, respectively. The first reagent-carrying chamber 211a contains a lysate, the second reagent-carrying chamber 211b contains a binding solution, the third reagent-carrying chamber 211c contains magnetic beads, the fourth reagent-carrying chamber 211d contains a cleaning solution A, the fifth reagent-carrying chamber 211e contains a cleaning solution B, and the sixth reagent-carrying chamber 211f contains an elution solution.

[0058] Among them, the portable instrument matched with the nucleic acid extraction device should be equipped with a power source for driving the piston 41, which is used to squeeze the reagent carrying chamber and the downward pressure rod of the elastic membrane 31 above the microvalve and the electromagnet for controlling the magnetic beads.

[0059] The specific implementation is described as follows:

[0060] A liquid sample (saliva, processed sputum, blood, swab rinse solution, etc.) or a swab is added into the sample elution chamber 35 through the sample adding port 34, the sealing sticker 11 is pressed tightly, and the nucleic acid extraction device is placed in a matching portable instrument.

[0061] After the portable instrument is started, the pressing rod moves downward to close the first microvalve 322a, the second microvalve 322b and the third microvalve 322c respectively. The pressing rod presses down the first reagent carrying chamber 211a, and as the aluminum foil 22 is broken, the lysate enters the sample elution chamber 35 through the first reagent inlet 323a to lyse the sample.

[0062] After the sample lysis is completed, the second reagent carrying chamber 211b is pressed down to add the binding solution into the sample elution chamber 35. The first microvalve 322a is opened, and the piston 41 is driven to move outward to drive the liquid in the sample elution chamber 35 to flow into the reaction chamber 36. At the same time, the third reagent carrying chamber 211c is pressed down to flush the magnetic beads into the reaction chamber 36 by the liquid flow, and the nucleic acid in the sample lysis product is captured together with the binding solution.

[0063] After the nucleic acid is captured, the electromagnet at the bottom of the reaction chamber 36 is turned on to adsorb the magnetic beads at the bottom of the reaction chamber 36, and the piston 41 is driven to move outward to drive the liquid in the reaction chamber 36 into the waste liquid chamber 37. The waste liquid chamber 37 has a built-in absorbent material that can absorb waste liquid to prevent contamination.

[0064] Next, the nucleic acid in the reaction chamber 36 is cleaned for the first time. The first microvalve 322a is closed, the electromagnet at the bottom of the reaction chamber 36 is disconnected, the fourth reagent carrying chamber 211d is pressed down to add the cleaning liquid A into the reaction chamber 36, the electromagnet at the bottom of the reaction chamber 36 is turned on, the magnetic beads are adsorbed on the bottom of the reaction chamber 36, and the driving gear 43 drives the piston 41 to move outward to drive the liquid in the reaction chamber 36 into the waste liquid chamber 37.

[0065] Then, the nucleic acid in the reaction chamber 36 is cleaned for the second time. The electromagnet at the bottom of the reaction chamber 36 is disconnected, and the fifth reagent carrying chamber 211e is pressed down to add the cleaning solution B into the reaction chamber 36. The electromagnet at the bottom of the reaction chamber 36 is turned on to adsorb the magnetic beads at the bottom of the reaction chamber 36. The piston 41 is driven to move outward to drive the liquid in the reaction chamber 36 into the waste liquid chamber 37. Through this operation, the impurities in the reaction chamber 36 except the nucleic acid adsorbed on the magnetic beads are washed away.

[0066] After the nucleic acid is cleaned, the electromagnet at the bottom of the reaction chamber 36 is disconnected, and the sixth reagent carrying chamber 211 f is pressed down to add the eluent into the reaction chamber 36 .

[0067] After a certain period of time, the electromagnet at the bottom of the reaction chamber 36 is turned on to adsorb the magnetic beads at the bottom of the reaction chamber 36, close the fourth microvalve 322d, and open the second microvalve 322b and the third microvalve 322c. The driving piston 41 moves outward to drive the liquid in the reaction chamber 36 to enter the downstream nucleic acid analysis device through the nucleic acid outlet 39. In this process, the two channels in the connection interface 38, one for discharging the extracted nucleic acid and the other for evacuating the gas in the nucleic acid analysis device, ensure a fully sealed operation.

[0068] Embodiment 2

[0069] When equipped with rapid membrane extraction technology, the nucleic acid extraction device is similar to that of the first embodiment, except that:

[0070] Only five reagent-carrying chambers are needed, and the shape of the reagent-carrying chambers is circular, and the diameter is 12 mm. The first reagent-carrying chamber 211a contains a digestion buffer, the second reagent-carrying chamber 211b contains ethanol, the third reagent-carrying chamber 211c contains a rinse solution A, the fourth reagent-carrying chamber 211d contains a rinse solution B, and the fifth reagent-carrying chamber 211e contains an elution solution. The reaction chamber 36 contains an enrichment membrane for capturing nucleic acids.

[0071] Among them, the portable instrument matched with the nucleic acid extraction device should be equipped with a power source for driving the piston 41, a pressing rod for squeezing the reagent carrying chamber and the elastic membrane 31 above the microvalve, and a local heating module for heating the sample elution chamber 35.

[0072] The specific implementation is described as follows:

[0073] The sample adding method is the same as that in the first embodiment, and will not be described in detail. It should be noted that the sample elution chamber 35 has proteinase K built in and a local heating module externally installed.

[0074] After the portable instrument is started, the pressing rod moves downward to close the first microvalve 322a, the second microvalve 322b and the third microvalve 322c respectively. The pressing rod presses down the first reagent carrying chamber 211a, and as the aluminum foil 22 is broken, the digestion buffer is transferred to the sample elution chamber 35 to mix with the sample and proteinase K, and incubated at a certain temperature for 10 minutes to complete the cleavage of the sample and expose the DNA.

[0075] After the sample lysis is completed, the second reagent carrying chamber 211b is pressed down, and ethanol is added to the sample elution chamber 35 to capture the water molecules around the DNA and precipitate the DNA.

[0076] Then, the first microvalve 322a is opened, and the piston 41 is driven to move outward to drive the liquid in the sample elution chamber 35 into the waste liquid chamber 37 through the reaction chamber 36. The reaction chamber 36 is equipped with an enrichment membrane to filter and adsorb DNA in the sample lysis product. The waste liquid chamber 37 is equipped with a water-absorbing material to absorb waste liquid and prevent contamination.

[0077] After the nucleic acid adsorption is completed, the first microvalve 322a is closed, and while the third reagent carrying chamber 211c is pressed down, the piston 41 is driven to move outward to drive the rinsing liquid A through the reaction chamber 36 into the waste liquid chamber 37, and the impurities other than the DNA adsorbed on the enrichment membrane in the reaction chamber 36 are washed away. Next, while the fourth reagent carrying chamber 211d is pressed down, the piston 41 is driven to move outward to drive the rinsing liquid B through the reaction chamber 36 into the waste liquid chamber 37, and the impurities other than the DNA adsorbed on the enrichment membrane in the reaction chamber 36 are washed away again.

[0078] After the nucleic acid rinse is completed, the fourth microvalve 322d is closed, and the second microvalve 322b and the third microvalve 322c are opened. While pressing down the fifth reagent carrying chamber 211e, the driving piston 41 moves outward to drive the liquid in the reaction chamber 36 to enter the downstream nucleic acid analysis device through the interface.

[0079] In summary, the nucleic acid extraction device in the present application, the housing 1, the reagent storage unit 2, the extraction operation unit 3 and the control layer 4, after being assembled, form a universal automated nucleic acid extraction device based on microfluidics and microvalves. The reagent storage unit 2 with built-in reagents is located above the extraction operation unit 3, and the reagent carrying chamber corresponds to the reagent inlet one by one through the aluminum foil 22; the control layer 4 is located below the extraction operation unit 3, and the piston 41 is placed at the deepest part of the drive chamber 42; the housing 1 covers the reagent storage unit 2, the extraction operation unit 3 and the control layer 4, forming a complete and sealed independent module.

[0080] The nucleic acid extraction device in the present application, when in use, opens the sealing patch 11, adds or rinses the sample into the sample elution chamber 35, and closes the sealing patch 11; external force presses down the reagent carrying chamber to break the aluminum foil 22 to release the reagent; by breaking the aluminum foil 22 under different reagent carrying chambers and switching different microvalves, different reagents flow into the reaction chamber 36 and the waste liquid chamber 37 in different orders; finally, the extracted nucleic acid is injected into various downstream nucleic acid analysis ends through the connection interface 38.

[0081] The nucleic acid extraction device in the present application, relying on the full system closure, full process integration, and full system automation of nucleic acid extraction, can provide effective support for a variety of nucleic acid analysis terminals.

[0082] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled 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 list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A nucleic acid extraction device, characterized in that: At least: A reagent storage unit (2) having a plurality of sealed reagent carrying chambers for carrying different reagents; The extraction operation unit (3) has a sample elution chamber (35) and a reaction chamber (36) connected to each other, wherein a part of the reagent carrying chamber is connected to the sample elution chamber, and another part of the reagent carrying chamber is connected to the reaction chamber (36); the sample elution chamber (35) has a sample injection port (34) connected to the external environment; A sealing patch (11) is arranged at the sample adding port (34). The sealing patch (11) is opened when adding samples, and the sealing patch (11) re-seals the sample adding port (34) after adding samples. The reagent storage unit (2) comprises a stacked storage substrate and an aluminum foil (22); The storage substrate is provided with a plurality of blisters (21) protruding in a direction away from the aluminum foil (22), and the space between the blisters (21) and the aluminum foil (22) is used to form the reagent carrying chamber; It also comprises a housing (1), wherein the housing (1) comprises an upper cover (13) and a lower cover (14) which are connected to each other; The sample adding port (34) is connected to the external environment through the upper cover (13); The upper cover (13) is provided with a first control channel (15) leading to the bubble cap (21), and different bubbles (21) are squeezed through different first control channels (15) to make the reagent in the bubble cap (21) flow out; It also includes a sticker (12) which is arranged on the surface of the upper cover (13) and is used to encapsulate the first control channel (15) on the upper cover (13).

2. The nucleic acid extraction device according to claim 1, characterized in that: The extraction operation unit (3) comprises a first fluid layer (32) and a second fluid layer (33) which are stacked, and the first fluid layer (32) is arranged in contact with the aluminum foil (22); The first fluid layer (32) is provided with a plurality of reagent inlets corresponding one to one with the reagent carrying chambers; The sample elution chamber (35) and the reaction chamber (36) are arranged in the second fluid layer (33); A portion of the reagent inlet is communicated with the sample elution chamber (35), and another portion of the reagent inlet is communicated with the reaction chamber (36).

3. The nucleic acid extraction device according to claim 2, characterized in that: The extraction operation unit (3) further comprises a waste liquid chamber (37) which is arranged in the second fluid layer (33), and the waste liquid chamber (37) is communicated with the reaction chamber (36).

4. The nucleic acid extraction device according to claim 3, characterized in that: It also includes a control layer (4) disposed on a side of the second fluid layer (33) facing away from the first fluid layer (32); A driving chamber (42) is provided in the control layer (4), and the driving chamber (42) is communicated with the waste liquid chamber (37); A piston (41) is provided in the driving chamber (42). When the piston (41) moves in the driving chamber (42), it is used to form a negative pressure in the driving chamber (42) to drive the reagent in the reagent-carrying chamber to flow into the sample elution chamber (35) or the reaction chamber (36), the liquid in the sample elution chamber (35) to flow into the reaction chamber (36), and the liquid in the reaction chamber (36) to flow into the waste liquid chamber (37).

5. The nucleic acid extraction device according to claim 4, characterized in that: The control layer (4) is also provided with a connection interface (38) for connecting the nucleic acid extraction device with the nucleic acid analysis device; The connection interface (38) is provided with a gas inlet (40) and a nucleic acid outlet (39); The gas inlet (40) and the nucleic acid outlet (39) are both connected to the reaction chamber (36).

6. The nucleic acid extraction device according to claim 5, characterized in that: Microvalves are provided between the sample elution chamber (35) and the reaction chamber (36), between the reaction chamber (36) and the nucleic acid outlet (39), between the reaction chamber (36) and the gas inlet (40), and between the reaction chamber (36) and the waste liquid chamber (37).

7. The nucleic acid extraction device according to claim 6, characterized in that: The space between the upper cover (13) and the lower cover (14) is used to accommodate the reagent storage unit (2), the extraction operation unit (3) and the control layer (4).

8. The nucleic acid extraction device according to claim 7, characterized in that: A first flow hole (52) and a second flow hole (53) for connecting the upstream and downstream of the microvalve are arranged in the valve cavity (51) of the microvalve; The valve cavity (51) of the microvalve is an open structure, and an elastic membrane (31) is provided at the open portion of the valve cavity (51). When the elastic membrane (31) is pressed downward, the first flow hole (52) and the second flow hole (53) are blocked; The upper cover (13) is provided with a second control channel (16) leading to the valve chamber (51), and different elastic membranes (31) are squeezed through different second control channels (16) to control the opening or closing of different microvalves.

9. The nucleic acid extraction device according to claim 8, characterized in that: The sticker (12) is also used to seal the second control channel (16) on the upper cover (13).

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