Nucleic acid extraction device including a mixing device

Abstract

The present invention relates to a nucleic acid extraction device including a mixing device, which includes a sample chamber, a sample chamber cover, a filtering component, a reaction chamber, a reaction chamber cover, and a mixing device. The nucleic acid extraction device of the present invention can achieve high-throughput filtration, reduce manual operations after adding a sample to be extracted, improve efficiency, and also enable highly automated continuous nucleic acid extraction and reaction, and can achieve efficient mixing of materials to avoid contamination.

Description

Technical Field

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

[0002] Nucleic acid is a macromolecule consisting of multiple nucleotides, which is widely present in all animal cells, plant cells and microorganisms. Nucleic acid is the genetic material of all organisms, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). It mainly exists in the cell nucleus and exists in a state of binding to proteins. Nucleic acid extraction is a very important technology in life science research, biotechnology application and genetic diagnosis. With the rapid development of molecular biology technology, the research and analysis of nucleic acids has been continuously promoted and applied in clinical diagnosis, food safety, environmental testing, agriculture, forestry and animal husbandry and other fields.

[0003] In addition to nucleic acids, biological cells also contain other substances such as proteins and polysaccharides. To analyze and study nucleic acids, it is first necessary to extract high-purity nucleic acids from complex biological environments. Extracting nucleic acids mainly includes lysis, adsorption, washing and desorption.

[0004] At present, commercial nucleic acid extraction and purification methods mainly include centrifugal column method and magnetic bead method. In the centrifugal column method, silicon-containing materials or other solids that can absorb nucleic acids are used as media to fill the centrifugal column, and chaotropic salts such as guanidine salts are used as binders. When nucleic acid molecules pass through the centrifugal column, they are adsorbed on the solid phase, and then impurities are washed away with an ethanol-containing solution, and finally the nucleic acids are eluted with a low-salt solution; this method is simple and convenient, with high extraction efficiency and good extraction effect. It is currently the most used nucleic acid purification method. The disadvantages are that it requires repeated centrifugation during the extraction process, is not convenient for high-throughput and automated operations, and samples are prone to cross-contamination. In the magnetic bead method, magnetic microspheres with silicon materials or other materials that can bind nucleic acids on the surface are used as solid phase carriers to separate DNA and RNA from samples under the action of chaotropic salts (guanidine hydrochloride, guanidine isothiocyanate, etc.) and an external magnetic field; this method is simple to operate and easy to realize automation and high-throughput operations. The disadvantages are that the cost is high, the extraction efficiency is easily affected, and the extraction effect of magnetic beads varies greatly among different samples, different manufacturers, and different batches.

[0005] Many efforts have been made in this field to improve the method of purifying nucleic acids to achieve high-throughput and automated nucleic acid extraction. For example, the Chinese invention patent application 201910776442.8, entitled "A Trace DNA Extraction Capsule", previously submitted by the applicant, discloses a trace DNA extraction capsule, which includes an inner tube, an outer tube and a top cover, wherein the inner tube is piston-slidably arranged on the upper part of the outer tube, and the top cover is detachably covered on the top of the inner tube, characterized in that a receiving chamber is provided in the middle of the top cover, and a reaction reagent chamber is placed in the receiving chamber, a T-shaped push rod is slidably arranged on the upper part of the receiving chamber, a detachable push rod support sleeve is arranged between the push rod and the top of the top cover, a solid reaction reagent is installed in the reaction reagent chamber, a through hole is opened at the top of the reaction reagent chamber, and a tip is provided at the bottom end of the reaction reagent chamber; and a heat-sealing film is provided on the top cover, and the heat-sealing film is arranged below the tip. In the nucleic acid extraction capsule structure disclosed in the patent document, a solid reaction reagent can be placed in the reaction reagent cavity, and then the reaction reagent cavity is sealed inside the top cover by a heat-sealed film, and a liquid extraction reagent is placed in an inner tube and sealed by the top cover, so that the solid reaction reagent and the extraction reagent will not be contaminated before use, thereby improving the accuracy of the test. The disadvantage of the nucleic acid extraction device disclosed in the document is that the extraction process still needs to rely on centrifugation to separate and obtain the required nucleic acid sample.

[0006] The Chinese utility model patent 201821769808.6, entitled "A laboratory nucleic acid extraction instrument", discloses a laboratory nucleic acid extraction instrument, including a lysis device, an elution device, and a filtering device. The filtering device includes a housing chamber, a filter arranged in the housing chamber, and the filter includes a frame that is engaged with the inner wall of the housing chamber and a plurality of filter screens installed side by side on the frame. A first water inlet and a first water outlet are provided on one side wall of the housing chamber parallel to the filter screen, and a second water inlet and a second water outlet are provided on the other parallel side wall. The first water outlet and the second water outlet are respectively provided with a suction device, which can not only make the filtered solution pass through the filter screen better, but also wash and dry the filter device after completing the nucleic acid extraction work. In the nucleic acid extraction device disclosed in the patent document, by adjusting the number of filter screens, it can be applied to nucleic acid operations with different contents, reduce the amount of reagents used, and improve work efficiency. The disadvantage of the nucleic acid extraction device disclosed in the document is that when absorbing the lysis solution or the elution solution, it is still an intermittent operation.

[0007] Chinese utility model patent application 201520838200.4, with the patent name "A Nucleic Acid Extraction Device", discloses a nucleic acid extraction device, which includes an adsorption tube having a first connection end and an adsorption end, an adsorption filter element arranged at the above-mentioned adsorption end and capable of adsorbing nucleic acid components in the sample to be processed, and a piston tube, wherein a piston is built in the piston tube and can move up and down along the inner wall of the piston tube, and the piston tube includes a second connection end and an open end and is connected to the first connection end through the second connection end to communicate with the adsorption tube, and is characterized in that it also includes a suction component for sucking liquid flowing through the adsorption filter element. The patent document also discloses a nucleic acid extraction device, which includes an adsorption tube having a first connection end and an adsorption end, an adsorption filter element arranged at the above-mentioned adsorption end and capable of adsorbing nucleic acid components in a sample to be processed, and a piston tube with a built-in piston, the piston tube includes a second connection end and an open end, and is connected to the first connection end through the second connection end to communicate with the adsorption tube, the open end is provided with a sealing ring, the piston is embedded in the sealing ring and can move up and down along the length direction of the piston tube, and is characterized in that it also includes a suction component for sucking liquid flowing through the adsorption filter element. In the nucleic acid extraction device disclosed in the patent document, the suction component preferably includes a straw, a one-way valve and a vacuum pump, one end of the straw is used to suck liquid, and the other end is connected to the vacuum pump through the one-way valve; or the absorption component includes a straw, an electric valve and an electric air pump, one end of the straw is used to suck liquid, and the other end is connected to the electric air pump through the electric valve. The disadvantage of the nucleic acid extraction device disclosed in the document is that the suction component sucks the filtrate after filtration, and does not realize the automatic extraction of nucleic acid samples.

[0008] In addition, when using the magnetic bead method to extract nucleic acids, a better extraction effect can only be achieved when the lysis reagent, magnetic beads and biological samples are fully mixed.

[0009] To this end, there is a continuous need in the art to develop a nucleic acid extraction device with high throughput, high degree of automation and good mixing effect. Summary of the invention

[0010] The purpose of the present invention is to overcome the defects of the above-mentioned prior art and provide a high-throughput, highly automated and efficient mixing nucleic acid extraction device.

[0011] In order to achieve the purpose of the present invention, this application provides the following technical solutions.

[0012] In one embodiment, the present application provides a nucleic acid extraction device including a mixing device, characterized in that the nucleic acid extraction device including the mixing device includes a sample chamber, a sample chamber cover, a filter assembly, a reaction chamber, a reaction chamber cover, and a mixing device;

[0013] Wherein, along the direction of fluid flow, the sample chamber includes at least one sample chamber inlet and at least one sample chamber outlet, the filter assembly includes at least one filter assembly inlet and at least one filter assembly outlet, the reaction chamber includes at least one reaction chamber inlet and at least one reaction chamber outlet, the sample chamber cover is arranged on the at least one sample chamber inlet, and the reaction chamber cover is arranged on the at least one reaction chamber outlet;

[0014] Wherein, the filter assembly can be switched between an initial state position and a transfer state position. When the filter assembly is in the initial state position, the sample chamber, the filter assembly and the reaction chamber are not fluidically connected; when the filter assembly is in the transfer state position, the sample chamber, the filter assembly and the reaction chamber are fluidically connected.

[0015] The sample chamber cover is provided with a hollow accommodating chamber surrounded by a fixing member and opening toward the sample chamber, a push rod extending above the sample chamber cover is slidably provided on the upper part of the hollow accommodating chamber, a movable reaction reagent chamber is provided in the middle part of the hollow accommodating chamber, and a sealing film is provided at the bottom of the hollow accommodating chamber, wherein the reaction reagent chamber is provided with a through hole, a tip is provided at the bottom of the reaction reagent chamber, and the sealing film is provided below the tip of the reaction reagent chamber;

[0016] The mixing device comprises a first reagent chamber for storing reagents, a second reagent chamber for storing reagents and the reaction chamber, the first reagent chamber is in fluid communication with the reaction chamber through a first opening, and the second reagent chamber is in fluid communication with the reaction chamber through a second opening;

[0017] wherein a first pierceable seal is provided at the first opening;

[0018] Wherein, along the direction toward the reaction chamber, the first reagent chamber sequentially comprises a first sealed reagent chamber and a second sealed reagent chamber separated by a second pierceable seal, the first opening is arranged in the second sealed reagent chamber, and the second sealed reagent chamber comprises a first piercing mechanism and a second piercing mechanism arranged therein, the first piercing mechanism is used to pierce the first pierceable seal, and the second piercing mechanism is used to pierce the second pierceable seal;

[0019] Wherein, along the direction toward the reaction chamber, the second reagent chamber comprises in sequence a third sealed reagent chamber and a sealed mixing chamber separated by a third pierceable seal, the second opening is arranged in the mixing chamber, and the mixing chamber comprises a third piercing mechanism arranged therein, and the third piercing mechanism is used to pierce the third pierceable seal;

[0020] Wherein, the first sealed reagent chamber cooperates with the second piercing mechanism;

[0021] Wherein, the third sealed reagent chamber cooperates with the third piercing mechanism.

[0022] In one embodiment of the first aspect, the filter assembly includes a filter assembly body and a switch mechanism, wherein the filter assembly body includes a pressing ring, a filter membrane and a hollow fluid channel sequentially arranged along a fluid flow direction, an inlet of the pressing ring is the at least one filter assembly inlet, and an outlet of the hollow flow channel is the at least one filter assembly outlet;

[0023] The switch mechanism is used to realize the reversible switching of the filter assembly between the initial state position and the transfer state position.

[0024] In one embodiment of the first aspect, the hollow fluid channel is arranged obliquely, wherein an end of the fluid channel close to the sample chamber is higher than an end of the fluid channel close to the reaction chamber.

[0025] In one implementation of the first aspect, the radial dimension of the lower portion of the sample chamber gradually decreases, and the minimum radial dimension of the lower portion of the sample chamber is smaller than the minimum radial dimension of the reaction reagent chamber.

[0026] In one embodiment of the first aspect, the first reagent chamber is disposed above the second reagent chamber, and the sample chamber is disposed above the first reagent chamber.

[0027] In one embodiment of the first aspect, the first piercing mechanism is disposed above the second piercing mechanism.

[0028] In one embodiment of the first aspect, the first piercing mechanism is a tapered piercing mechanism with a pointed end directed toward the first pierceable seal and extending a first length.

[0029] In one embodiment of the first aspect, the first length of the first piercing mechanism is equal to a length of the second sealed reagent chamber extending in a direction toward the reaction chamber.

[0030] In one embodiment of the first aspect, the second piercing mechanism is a conical piercing mechanism with a tip directed toward the second pierceable seal and extending a second length, and the second length of the second piercing mechanism is less than the first length of the first piercing mechanism.

[0031] In one embodiment of the first aspect, the tip end face of the second piercing mechanism includes a seal piercing area and a seal pressing area, wherein the seal piercing area is closer to the second pierceable seal than the seal pressing area, and along the direction toward the second pierceable seal, the length of the bottom of the seal pressing area is greater than the length of the top of the seal pressing area.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (1) By providing a filter component that can be switched between an initial state position and a transfer state position in the nucleic acid extraction device, the fluid connection relationship between the sample chamber and the reaction chamber is cleverly controlled to achieve high-throughput filtration;

[0034] (2) By pre-loading at least a portion of the reaction reagent in the reaction reagent chamber and the sample chamber, the manual operation after adding the sample to be extracted can be reduced, thereby improving efficiency;

[0035] (3) By connecting the outlet of the reaction chamber to a negative pressure device, the filtration efficiency and processing throughput are further improved, and nucleic acid extraction and reaction can be performed continuously and in a highly automated manner;

[0036] (4) By cleverly setting up multiple sealed reagent chambers and piercing mechanisms, efficient mixing of materials can be achieved to avoid contamination. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 4 is a cross-sectional view of a nucleic acid extraction device including a mixing device according to an embodiment of the present invention in an initial state.

[0038] Figure 2 It is a cross-sectional view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a sample loading state.

[0039] Figure 3 4 is a cross-sectional view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a transfer state.

[0040] Figure 4 It is a stereoscopic view of a second piercing mechanism according to an embodiment of the present invention.

[0041] Figure 5 It is a left side view of a mixing device according to one embodiment of the present invention.

[0042] Figure 6 It is a front view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention in an initial state.

[0043] Figure 7It is a front view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a sample loading state.

[0044] In the above drawings, the meanings of the reference numerals are as follows:

[0045] 1000 Nucleic acid extraction device including a mixing device

[0046] 100 sample chamber cover

[0047] 101 Fixings

[0048] 102 Putter

[0049] 103 Accommodation cavity

[0050] 104 Reaction chamber

[0051] 105 Sealing Film

[0052] 106 sealing ring

[0053] 200 sample chambers

[0054] 201 Sample chamber entrance

[0055] 202 Sample chamber outlet

[0056] 203 Funnel-type diversion device

[0057] 300 filter components

[0058] 301 filter component entrance

[0059] 302 filter component outlet

[0060] 303 pressure ring

[0061] 304 filter membrane

[0062] 305 Hollow Fluid Channel

[0063] 306 switch mechanism

[0064] 400 reaction chamber

[0065] 401 reaction chamber entrance

[0066] 402 reaction chamber outlet

[0067] 403 First Interface

[0068] 404 first interface sealing ring

[0069] 500 reaction chamber cover

[0070] 601 Sample Chamber Heater

[0071] 602 reaction chamber heater

[0072] 700 reaction chamber sealing ring

[0073] 800 First Reagent Chamber

[0074] 801 first sealed reagent tank

[0075] 802 Second pierceable seal

[0076] 803 Second piercing mechanism

[0077] 8031 Seal puncture area

[0078] 8032 seal extrusion area

[0079] 804 Second sealed reagent compartment

[0080] 805 First pierceable seal

[0081] 806 first piercing mechanism

[0082] 807 First Opening

[0083] 808 First Piston

[0084] 809 first piston hook

[0085] 900 Second reagent chamber

[0086] 901 Third sealed reagent compartment

[0087] 902 Third pierceable seal

[0088] 903 Third piercing mechanism

[0089] 904 Mixing Chamber

[0090] 905 Second Opening

[0091] 906 Second Piston

[0092] 907 Second piston hook. DETAILED DESCRIPTION

[0093] Unless otherwise defined, the technical terms or scientific terms used in this specification and claims shall be the ordinary meanings understood by persons with ordinary skills in the technical field to which the present invention belongs. All the values ​​from the lowest value to the highest value listed herein refer to all the values ​​obtained by incrementing the lowest value and the highest value by one unit when the difference between the lowest value and the highest value is more than two units. In the description of the invention, it should be understood that the orientation or position relationship indicated by the terms "center", "longitudinal", "lateral", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. is based on the orientation or position relationship shown in the accompanying drawings, which is only for the convenience of describing the 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 a limitation on the invention. In addition, the terms "first", "second", etc. are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, the features defined as "first", "second", etc. may explicitly or implicitly include one or more of the features. In the description of the present invention, unless otherwise specified, “plurality” means two or more.

[0094] In the description of the invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installation", "connection" and "connection" 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 invention can be understood according to specific circumstances.

[0095] The specific embodiments of the present invention will be described below in conjunction with the accompanying drawings. It should be noted that in the specific description of these embodiments, for the sake of concise description, this specification cannot provide a detailed description of all the features of the actual embodiments. Without departing from the spirit and scope of the present invention, those skilled in the art may modify and replace the embodiments of the present invention, and the resulting embodiments are also within the scope of protection of the present invention.

[0096] Definition of terms

[0097] Herein, the term "initial state position" refers to the position of the filtering component when the nucleic acid extraction device including the mixing device is in the initial state.

[0098] As used herein, the term "transition state position" refers to the position of the filtering component when the nucleic acid extraction device including the mixing device is in the transition state.

[0099] As mentioned above, the existing nucleic acid extraction device including a mixing device is difficult to achieve high throughput and automated operation. The purpose of this application is first to provide a nucleic acid extraction device including a mixing device that can achieve high throughput, automated operation and high mixing efficiency.

[0100] In one embodiment, the nucleic acid extraction device including a mixing device described herein may include a sample chamber, a sample chamber cover, a filter assembly, a reaction chamber, and a reaction chamber cover. In this embodiment, along the direction of fluid flow, the sample chamber includes at least one sample chamber inlet and at least one sample chamber outlet, the filter assembly includes at least one filter assembly inlet and at least one filter assembly outlet, the reaction chamber includes at least one reaction chamber inlet and at least one reaction chamber outlet, the sample chamber cover is disposed above the at least one sample chamber inlet, and the reaction chamber cover is disposed above the at least one reaction chamber outlet. Those skilled in the art can understand that as long as reasonable fluid flow can be achieved and the sealing of the nucleic acid extraction device including a mixing device can be ensured, the sample chamber, the filter assembly, and the reaction chamber can each independently include one or more inlets and outlets.

[0101] In this embodiment, the filter assembly can switch between an initial state position and a transfer state position. When the filter assembly is in the initial state position, the sample chamber, the filter assembly and the reaction chamber are not fluidically connected; when the filter assembly is in the transfer state position, the sample chamber, the filter assembly and the reaction chamber are fluidically connected.

[0102] In this embodiment, the sample chamber cover is provided with a hollow accommodating chamber surrounded by a fixing member and opening toward the sample chamber, a push rod extending above the sample chamber cover is slidably provided on the upper part of the hollow accommodating chamber, a movable reaction reagent chamber is provided in the middle part of the hollow accommodating chamber, and a sealing film is provided at the bottom of the hollow accommodating chamber, wherein the reaction reagent chamber is provided with a through hole, a tip is provided at the bottom of the reaction reagent chamber, and the sealing film is provided below the tip of the reaction reagent chamber. For more descriptions of the reaction reagent chamber, please refer to the Chinese invention patent application previously submitted by the applicant, which is entitled "A Trace DNA Extraction Capsule" and has an application number of 201910776442.8.

[0103] In the embodiments described herein, the sample chamber is mainly used to lyse biological samples containing nucleic acids, such as cotton swab wipes / adhesions / adsorbates containing exfoliated cells, etc. In one embodiment, solid reagents can be pre-set in the reaction reagent chamber of the sample chamber cover, and liquid reagents can be pre-set in the sample chamber at the same time. After adding the biological sample to the sample chamber, the lysis reaction can be started by pressing the push rod set in the sample chamber cover. There is no need to add additional reaction reagents, which can significantly reduce the workload of on-site manual operations, shorten the nucleic acid extraction time, and thus improve efficiency. In addition, because the nucleic acid extraction device including the mixing device is sealed during the nucleic acid extraction process, the risk of contamination of the biological sample is reduced.

[0104] In the embodiments described herein, the filter assembly is mainly used to control the flow of fluid between the sample chamber and the reaction chamber. When the filter assembly is in the initial state position, that is, when filtering is not required, a passage cannot be formed between the sample chamber, the filter assembly and the reaction chamber. At this time, the material in the sample chamber cannot flow into the reaction chamber. It can be understood by those skilled in the art that as long as there is at least one place between the sample chamber and the filter assembly or between the filter assembly and the reaction chamber without fluid communication, a passage cannot be formed between the sample chamber, the filter assembly and the reaction chamber. Similarly, when the filter assembly is in the transfer state position, that is, when filtering is required, a passage is formed between the sample chamber, the filter assembly and the reaction chamber, and the material in the sample chamber, especially the liquid material, can flow into the reaction chamber for subsequent reaction. Although the switching of the initial state position and the transfer state position of the filter assembly is realized in the form of a sliding switch mechanism below, it can be understood by those skilled in the art that other devices and methods can also be used to realize such switching, and these devices and methods are within the scope of protection of the present application. For example, in one embodiment, the mechanical position of the filter assembly may not change, and a plug is set on at least one of the sample chamber outlet, the filter assembly inlet, the filter assembly outlet and the sample chamber inlet. When the filter assembly is in the initial state position, that is, when filtering is not required, at least one plug blocks at least one corresponding outlet or inlet. When the filter assembly is in the transfer state position, that is, when filtering is required, the plugs at all outlets and inlets are removed.

[0105] In the embodiments described herein, the reaction chamber is used to adsorb the nucleic acid sample after lysis onto the magnetic beads. The magnetic beads adsorbed with the nucleic acid sample can be transferred to an external device through an interface provided in the reaction chamber.

[0106] The method of use and working principle of the nucleic acid extraction device including a mixing device described herein are as follows. In summary, the nucleic acid extraction device including a mixing device described herein may include an initial state, a sample loading state, and a transfer state. Before use, the nucleic acid extraction device including a mixing device is in an initial state. When it is necessary to extract a nucleic acid sample, the sample to be extracted first needs to be added to the sample chamber of the nucleic acid extraction device including a mixing device, and at this time, the nucleic acid extraction device including the mixing device is in a sample loading state. When the cleavage reaction is completed and the nucleic acid sample after cleavage needs to be transferred to the reaction chamber, the nucleic acid extraction device including the mixing device is in a transfer state.

[0107] When the nucleic acid extraction device including the mixing device is in the initial state, the sample chamber cover is covered on the sample chamber, the reaction chamber cover is covered on the reaction chamber, and the filter assembly is in the initial state position. At this time, the sample chamber, the filter assembly and the reaction chamber are not fluidically connected. The reaction reagent chamber of the sample chamber cover is pre-set with a solid reagent for a cleavage reaction, and the sample chamber is pre-set with a liquid reagent for a cleavage reaction.

[0108] When the nucleic acid extraction device including the mixing device is in the sample loading state, the push rod in the sample chamber cover is first pressed downward to move the reaction reagent chamber downward, and the tip of the reaction reagent chamber pierces the sealing film arranged below it and falls into the sample chamber. Then, the sample chamber cover is opened, and a biological sample containing nucleic acid, such as a wiping cotton swab containing exfoliated cells, is put into the sample chamber, and the sample chamber cover is closed, and the lysis reaction begins.

[0109] After the cleavage reaction is completed, the nucleic acid extraction device including the mixing device can be switched to the transfer state. At this time, the reaction chamber cover is opened, and the reaction chamber outlet is connected to the negative pressure device. Then, the filter component is switched to the transfer state position, so that the cleaved material is filtered through the filter component and then enters the reaction chamber to complete the nucleic acid sample extraction.

[0110] The mixing device of the nucleic acid extraction device including the mixing device described herein includes a plurality of sealed reagent chambers and reaction chambers, wherein the sealed reagent chambers are sealed by pierceable seals and can be pierced by piercing mechanisms, thereby mixing the materials stored in the sealed reagent chambers. More detailed features of the mixing device will be described below.

[0111] Example 1

[0112] This embodiment relates to a nucleic acid extraction device including a mixing device according to one embodiment of the present invention.

[0113] The nucleic acid extraction device 1000 including the mixing device of this embodiment will be described in detail below with reference to the accompanying drawings. Figure 1 and Figure 6The nucleic acid extraction device 1000 including the mixing device is described in an initial state. Figure 1 4 is a cross-sectional view of a nucleic acid extraction device including a mixing device according to an embodiment of the present invention in an initial state. Figure 6 1 is a front view of a nucleic acid extraction device including a mixing device according to an embodiment of the present invention in an initial state. The nucleic acid extraction device including a mixing device 1000 described herein can be used in combination with an external device. For more description of the external device, please refer to the Chinese invention patent application with publication number CN105950438A and title “A reagent cartridge provided with a press-type magnetic bead transfer mechanism”.

[0114] like Figure 1 and Figure 6 As shown, the nucleic acid extraction device 1000 described in this embodiment may include a sample chamber cover 100, a sample chamber 200, a filter assembly 300, a reaction chamber 400 and a reaction chamber cover 500. The sample chamber cover 100 may include a hollow accommodating chamber 103 surrounded by a fixing member 101 and opening toward the sample chamber 200, a push rod 102 extending above the sample chamber cover 100 is slidably provided at the upper part of the hollow accommodating chamber 103, a movable reaction reagent chamber 104 is provided in the middle of the hollow accommodating chamber 103, and a sealing film 105 is provided at the bottom of the hollow accommodating chamber 103, wherein the reaction reagent chamber 104 is provided with a through hole, the bottom of the reaction reagent chamber 104 is provided with a tip, and the sealing film 105 is provided below the tip of the reaction reagent chamber.

[0115] In this embodiment, the reaction reagent chamber 104 is made of a material having a density greater than that of the liquid reaction reagent in the sample chamber 200, so that when the reaction reagent chamber 104 falls into the sample chamber 200, it will sink to the bottom instead of floating above the liquid surface. In this embodiment, a through hole can be set on the top, side or bottom of the reaction reagent chamber 104, so that when the reaction reagent chamber 104 falls into the sample chamber 200, the liquid reagent can be immersed in the reaction reagent chamber 104 to dissolve the solid reaction reagent. In addition, in this embodiment, the through hole diameter of the reaction reagent chamber 104 should be smaller than the size of the solid reaction reagent pre-set therein, so as to prevent the solid reaction reagent from leaking into the hollow accommodating chamber 103 of the sample chamber cover 100 during storage and transportation.

[0116] In this embodiment, after the push rod 102 is pressed downward, the reaction reagent chamber 104 can pierce the sealing film 105 downward and fall into the sample chamber 200. In order to improve the sealing performance, a sealing ring 106 can be provided between the push rod 102 and the fixing member 101.

[0117] Also like Figure 1As shown, in the nucleic acid extraction device including the mixing device described in this embodiment, the sample chamber 200 may include a sample chamber inlet 201 and a sample chamber outlet 202. The radial dimension of the lower portion of the sample chamber 200 gradually decreases, and the minimum radial dimension of the lower portion of the sample chamber is smaller than the minimum radial dimension of the reaction reagent chamber 104. This arrangement is to buffer the vibration generated by the reaction reagent chamber 104 falling into the reaction reagent chamber 104 and prevent the reaction reagent chamber 104 from damaging the filter assembly.

[0118] Also like Figure 1 As shown, in the nucleic acid extraction device including the mixing device described in this embodiment, the filter assembly 300 may include a filter assembly inlet 301 and a filter assembly outlet 302. In this embodiment, the filter assembly 300 includes a filter assembly body and a switch mechanism 306. The filter assembly body includes a pressing ring 303, a filter membrane 304 and a hollow fluid channel 305 arranged in sequence along the fluid flow direction, the inlet of the pressing ring 303 is the filter assembly inlet 301, and the outlet of the hollow fluid channel 305 is the filter assembly outlet 302.

[0119] In this embodiment, the switch mechanism 306 is connected to the filter assembly body, and can make the filter assembly body slide horizontally left and right relative to the sample chamber 200. When the filter assembly 300 is in the initial state position, the filter assembly inlet 301 is not fluidically connected to the sample chamber outlet 202. When the filter assembly 300 is in the transfer state position, the filter assembly inlet 301 is fluidically connected to the sample chamber outlet 202. Sliding the switch mechanism 306 to the right, driving the filter assembly body to slide to the right together, can realize the switching of the filter assembly 300 from the initial state position to the transfer state position.

[0120] In this embodiment, the hollow fluid channel is arranged obliquely, wherein the end of the hollow fluid channel close to the sample chamber is higher than the end of the hollow fluid channel close to the reaction chamber. In this embodiment, the pore size of the filter membrane may be 0.2-0.5 μm.

[0121] Also like Figure 1 As shown, in the nucleic acid extraction device including a mixing device described in this embodiment, the reaction chamber 400 may include a reaction chamber inlet 401 and a reaction chamber outlet 402. In this embodiment, in order to facilitate the subsequent extraction of negative pressure from the reaction chamber 400, the reaction chamber 400 also includes a first interface 403 and a first interface sealing ring 404 arranged above the reaction chamber outlet 402. In addition, the reaction chamber 400 also includes a second interface (not shown in the figure) for communicating with an external device to transfer the nucleic acid sample after magnetic bead adsorption to other devices. The second interface may be arranged below the reaction chamber inlet 401. Those skilled in the art will appreciate that neither the first interface 403 nor the second interface is necessary.

[0122] In this embodiment, the first interface 403 is used to output gas, and the second interface is used to output liquid and solid materials. In one embodiment, in order to prevent the materials in the reaction chamber 400 from being contaminated, a one-way valve or a gas-permeable and water-impermeable filter membrane can be set at the first interface 403 .

[0123] In this embodiment, magnetic beads may be pre-set in the reaction chamber 400. In order to ensure the airtightness of the reaction chamber 400, a reaction chamber sealing ring 700 may be further provided at the junction between the reaction chamber 400 and other components.

[0124] like Figure 1 As shown, when the nucleic acid extraction device is in the initial state, the filter assembly 300 is in the initial state position, and the sample chamber outlet 202 and the filter assembly inlet 301 are staggered and not connected.

[0125] Also refer to Figure 1 The nucleic acid extraction device 100 including a mixing device described herein includes a mixing device, wherein the mixing device includes a first reagent chamber 800 for storing reagents, a second reagent chamber 900 for storing reagents, and a reaction chamber 400 for mixing materials. In this embodiment, the first reagent chamber 800 is in fluid communication with the reaction chamber 400 through a first opening 807. The second reagent chamber 900 is in fluid communication with the reaction chamber 400 through a second opening 905.

[0126] In this embodiment, a first pierceable seal 805 is disposed at the first opening 807. In a specific embodiment, the first pierceable seal 805 may be a sealing film, which separates the first reagent chamber 800 from the reaction chamber 400. However, after the first pierceable seal 805 is pierced, the first reagent chamber 800 and the reaction chamber 400 are in fluid communication.

[0127] In this embodiment, along the direction toward the reaction chamber 400, the first reagent chamber 800 sequentially includes a first sealed reagent chamber 801 and a second sealed reagent chamber 804 separated by a second pierceable seal 802. A first opening 807 is provided in the second sealed reagent chamber 804. The second sealed reagent chamber 804 includes a first piercing mechanism 806 and a second piercing mechanism 803 provided therein, wherein the first piercing mechanism 806 is used to pierce the first pierceable seal 805, and the second piercing mechanism 803 is used to pierce the second pierceable seal 802.

[0128] In one embodiment, the first opening 807 may be a hole and is arranged above the second sealed reagent warehouse 804, with a height higher than the top of the first piercing mechanism 806. This arrangement allows the reagents in the first sealed reagent warehouse 801 and the second sealed reagent warehouse 804 to be fully mixed before entering the reaction chamber 400. The first sealed reagent warehouse 801 and the second sealed reagent warehouse 804 can be used independently to store solid reagents or liquid reagents. In order to make the material mixing more sufficient, the first sealed reagent warehouse 801 can cooperate with the second piercing mechanism 803, that is, the first sealed reagent warehouse 801 can be arranged to fit the second piercing mechanism 803. In other words, the shape volume of the first sealed reagent warehouse 801 and the shape volume of the second piercing mechanism 803 are almost the same. When the second piercing mechanism 803 completely pierces the second pierceable seal 802, the inner wall of the second piercing mechanism 803 and the first sealed reagent warehouse 801 is almost directly in contact with each other without gaps, thereby ensuring that there is no residual reagent in the first sealed reagent warehouse 801, avoiding waste of reagents.

[0129] In this embodiment, along the direction toward the reaction chamber, the second reagent chamber 900 sequentially includes a third sealed reagent compartment 901 and a sealed mixing chamber 904 separated by a third pierceable seal 902. The second opening 905 is disposed in the mixing chamber 904, and the mixing chamber 904 includes a third piercing mechanism 903 disposed therein, and the third piercing mechanism 903 is used to pierce the third pierceable seal 902.

[0130] In one embodiment, the mixing chamber 904 may be configured to include two second openings 905, one of which is close to the top of the mixing chamber 904 and the other is close to the bottom of the mixing chamber 904. In this way, when the third sealed reagent compartment 901 reciprocates laterally relative to the reaction chamber 400, the materials are mixed evenly by allowing the materials to continuously enter and exit the mixing chamber 904. Similarly, the third sealed reagent compartment 901 is configured to fit the third piercing mechanism 903. In other words, the shape and volume of the third sealed reagent compartment 901 and the shape and volume of the third piercing mechanism 903 are almost the same. When the third piercing mechanism 903 completely pierces the third pierceable seal 902, the inner walls of the third piercing mechanism 903 and the third sealed reagent compartment 901 are in direct contact with each other almost without gaps, thereby ensuring that there is no residual reagent in the third sealed reagent compartment 901 and avoiding waste of reagents.

[0131] In one embodiment, in order to improve mixing efficiency, the first reagent chamber 800 is disposed above the second reagent chamber 900 .

[0132] In one embodiment, the first piercing mechanism 806 is configured to pierce the first pierceable seal 805 when the first piercing mechanism 806 moves toward the reaction chamber 400. The second piercing mechanism 803 is configured to pierce the second pierceable seal 802 when the first piercing mechanism 806 moves toward the reaction chamber 400. In one embodiment, the first piercing mechanism 806 is a conical piercing mechanism with a tip, the tip of which is toward the first pierceable seal 805 and extends a first length. In one embodiment, the second piercing mechanism 803 is a conical piercing mechanism with a tip, the tip of which is toward the second pierceable seal 802 and extends a second length. Figure 1 It can be seen that the tip of the second piercing mechanism 803 is arranged on the left, while the tip of the first piercing mechanism 806 is arranged on the right.

[0133] In one embodiment, the first length of the first piercing mechanism 806 is equal to the length of the second sealed reagent compartment 804 extending in the direction toward the reaction chamber, and the second length of the second piercing mechanism is less than the first length of the first piercing mechanism. This specific structure ensures that when the first reagent chamber 800 moves toward the reaction chamber 400, the first piercing mechanism 806 first pierces the first pierceable seal 805, so that the second sealed reagent compartment 804 is connected to the reaction chamber fluid, and the second sealed reagent compartment 804 is depressurized. Then, the second piercing mechanism 803 pierces the second pierceable seal 802, so that the materials stored in the first sealed reagent compartment 801 and the second sealed reagent compartment 804 are mixed, and finally enter the reaction chamber 400 through the first opening 807, which significantly improves the safety of the device operation.

[0134] In one embodiment, the first piercing mechanism 806 is arranged above the second piercing mechanism 803. In this embodiment, the first opening 807 corresponding to the first piercing mechanism 806 must be arranged above the second piercing mechanism 803. When a liquid reagent is stored in the first sealed reagent warehouse 801 or the second sealed reagent warehouse 804, this structure is particularly preferred. Specifically, because of the influence of gravity, the liquid reagent has a tendency to flow downward. In the case where the second sealed reagent warehouse 804 stores a liquid reagent, if the first piercing mechanism 806 and the first opening 807 are arranged below the second piercing mechanism 803, then after the first piercing mechanism 806 pierces the first pierceable seal 805, the liquid reagent has no time to fully mix and enters the reaction chamber 400. And the structure according to this embodiment can increase the effective mixing time of the reagent stored in the first sealed reagent warehouse 801 or the second sealed reagent warehouse 804, thereby improving the mixing efficiency.

[0135] Next, refer to Figure 4, the tip end face of the second piercing mechanism 803 includes a seal piercing area 8031 ​​and a seal extrusion area 8032, wherein the seal piercing area 8031 ​​is closer to the second pierceable seal 802 than the seal extrusion area 8032. Preferably, along the direction toward the second pierceable seal 802, the length of the bottom of the seal extrusion area 8032 is greater than the length of the top of the seal extrusion area 8032. According to the second piercing mechanism 803 of this embodiment, the seal piercing area can first be used to pierce the second pierceable seal 802, and then the second pierceable seal 802 is shoveled upwards through the seal extrusion area 8032. This not only speeds up the speed of mixing the reagents in the first sealed reagent compartment 801 or the second sealed reagent compartment 804, but also avoids the debris of the second pierceable seal 802 from contaminating the reagent.

[0136] refer to Figure 2 and Figure 3 In one embodiment, the first reagent chamber 800 includes a slidably connected first piston mechanism 808, which is connected to the mixing device via a first piston hook 809, and the first piston mechanism 808 is configured to control the lateral movement of the first reagent chamber 800 relative to the reaction chamber 400.

[0137] In another embodiment, the third piercing mechanism 903 is an arc-shaped piercing mechanism with the tip facing the third pierceable seal 902, and the top of the arc-shaped piercing mechanism has a raised tip portion.

[0138] Similarly, the second reagent chamber 900 includes a slidably connected second piston mechanism 906 , which is connected to the mixing device via a second piston hook 907 , and the second piston mechanism is configured to control the lateral movement of the second reagent chamber 900 relative to the reaction chamber 400 .

[0139] Next, we will combine Figure 2 and Figure 7 The nucleic acid extraction device 1000 including the mixing device in the sample loading state is described. Figure 2 It is a cross-sectional view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a sample loading state. Figure 7 It is a front view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a sample loading state.

[0140] from Figure 2It can be seen that when the nucleic acid extraction device 1000 including the mixing device is in the sample loading state, the filter assembly 300 is still in the initial state position. However, the reaction reagent chamber 104 has fallen into the sample chamber 200. This can be achieved by pressing down the push rod 102 set in the sample chamber cover 100. The reaction reagent chamber 104 can be pre-set with solid reagents required for the cleavage reaction. The sample chamber 200 can be pre-set with liquid reagents required for the cleavage reaction. After adding the biological sample to be lysed, the cleavage reaction can be carried out. The degree of manual intervention is greatly reduced, and the risk of sample contamination is avoided.

[0141] Finally, combine Figure 3 To describe the nucleic acid extraction device 1000 including the mixing device in a working state. Figure 3 4 is a cross-sectional view of a nucleic acid extraction device including a mixing device according to one embodiment of the present invention when in a transfer state.

[0142] The filter assembly 300 can be switched from the initial state position to the transfer state position by sliding the switch mechanism 306 to the right.

[0143] like Figure 3 As shown, when the nucleic acid extraction device including the mixing device is in the transfer state, the sample chamber outlet 202 is fluidically connected to the filter assembly inlet 301, and the filter assembly outlet 302 is fluidically connected to the reaction chamber inlet 401. The reaction chamber outlet 402 of the reaction chamber 400 can be connected to the negative pressure device through the first interface 403. Thereby, the material after lysis in the sample chamber 200 enters the reaction chamber 400 through the filter assembly 300 with high throughput. It can be understood by those skilled in the art that in order to improve the sealing performance, a first interface sealing ring 404 can be provided around the first interface 403. In addition, it can be understood by those skilled in the art that when a negative pressure operation is required, the reaction chamber cover 500 needs to be removed first.

[0144] Example 2

[0145] The nucleic acid extraction device of this embodiment includes all the technical features of the nucleic acid extraction device of embodiment 1, and a funnel-shaped flow guide device 203 is further provided at the lower part of the sample chamber 200 to provide a more effective buffering effect. The funnel-shaped flow guide device 203 is provided below the liquid level of the liquid reaction reagent, and after the reaction reagent chamber 104 falls into the sample chamber 200, it is sufficient to allow the liquid reaction reagent to immerse in the reaction reagent chamber 104.

[0146] Example 3

[0147] The nucleic acid extraction device of this embodiment includes all the technical features of the nucleic acid extraction device of embodiment 1, and a sample chamber heater 601 is provided below the sample chamber 200, and a reaction chamber heater 602 is provided below the reaction chamber. Because the cleavage reaction usually needs to be carried out at a high temperature, providing the sample chamber heater 601 can ensure the cleavage reaction and make the instrument structure compact.

[0148] In a first aspect, the present application provides a nucleic acid extraction device including a mixing device, characterized in that the nucleic acid extraction device includes a sample chamber, a sample chamber cover, a filter assembly, a reaction chamber, a reaction chamber cover, and a mixing device;

[0149] Wherein, along the direction of fluid flow, the sample chamber includes at least one sample chamber inlet and at least one sample chamber outlet, the filter assembly includes at least one filter assembly inlet and at least one filter assembly outlet, the reaction chamber includes at least one reaction chamber inlet and at least one reaction chamber outlet, the sample chamber cover is arranged on the at least one sample chamber inlet, and the reaction chamber cover is arranged on the at least one reaction chamber outlet;

[0150] Wherein, the filter assembly can be switched between an initial state position and a transfer state position. When the filter assembly is in the initial state position, the sample chamber, the filter assembly and the reaction chamber are not fluidically connected; when the filter assembly is in the transfer state position, the sample chamber, the filter assembly and the reaction chamber are fluidically connected.

[0151] The sample chamber cover is provided with a hollow accommodating chamber surrounded by a fixing member and opening toward the sample chamber, a push rod extending above the sample chamber cover is slidably provided on the upper part of the hollow accommodating chamber, a movable reaction reagent chamber is provided in the middle part of the hollow accommodating chamber, and a sealing film is provided at the bottom of the hollow accommodating chamber, wherein the reaction reagent chamber is provided with a through hole, a tip is provided at the bottom of the reaction reagent chamber, and the sealing film is provided below the tip of the reaction reagent chamber;

[0152] Wherein, the mixing device comprises a first reagent chamber for storing reagents, a second reagent chamber for storing reagents and the reaction chamber, the first reagent chamber is in fluid communication with the reaction chamber through a first opening, and the second reagent chamber is in fluid communication with the reaction chamber through a second opening;

[0153] wherein a first pierceable seal is provided at the first opening;

[0154] Wherein, along the direction toward the reaction chamber, the first reagent chamber sequentially comprises a first sealed reagent chamber and a second sealed reagent chamber separated by a second pierceable seal, the first opening is arranged in the second sealed reagent chamber, and the second sealed reagent chamber comprises a first piercing mechanism and a second piercing mechanism arranged therein, the first piercing mechanism is used to pierce the first pierceable seal, and the second piercing mechanism is used to pierce the second pierceable seal;

[0155] Wherein, along the direction toward the reaction chamber, the second reagent chamber comprises in sequence a third sealed reagent chamber and a sealed mixing chamber separated by a third pierceable seal, the second opening is arranged in the mixing chamber, and the mixing chamber comprises a third piercing mechanism arranged therein, and the third piercing mechanism is used to pierce the third pierceable seal;

[0156] Wherein, the first sealed reagent chamber is suitable for snugly accommodating the second piercing mechanism;

[0157] Wherein, the third sealed reagent chamber is suitable for snugly accommodating the third piercing mechanism.

[0158] In one embodiment of the first aspect, the filter assembly includes a filter assembly body and a switch mechanism, wherein the filter assembly body includes a pressing ring, a filter membrane and a hollow fluid channel sequentially arranged along a fluid flow direction, an inlet of the pressing ring is the at least one filter assembly inlet, and an outlet of the hollow flow channel is the at least one filter assembly outlet;

[0159] The switch mechanism is used to realize the reversible switching of the filter assembly between the initial state position and the transfer state position.

[0160] In one embodiment of the first aspect, the hollow fluid channel is arranged obliquely, wherein an end of the fluid channel close to the sample chamber is higher than an end of the fluid channel close to the reaction chamber.

[0161] In one embodiment of the first aspect, the pore size of the filter membrane is 0.2-0.5 μm.

[0162] In one implementation of the first aspect, the radial dimension of the lower portion of the sample chamber gradually decreases, and the minimum radial dimension of the lower portion of the sample chamber is smaller than the minimum radial dimension of the reaction reagent chamber.

[0163] In one embodiment of the first aspect, a funnel-shaped flow guide device is included above the outlet of the at least one sample chamber.

[0164] In one embodiment of the first aspect, the method further comprises a heater for heating the sample chamber.

[0165] In one embodiment of the first aspect, at least one outlet of the reaction chamber is connected to a negative pressure device.

[0166] In one embodiment of the first aspect, a one-way valve or an air-permeable and water-impermeable filter membrane is provided on the pipeline between at least one outlet of the reaction chamber and the negative pressure device.

[0167] In one embodiment of the first aspect, the reaction chamber further comprises at least one interface for outputting materials, and the at least one interface is arranged below the at least one inlet of the reaction chamber.

[0168] In another embodiment, the first reagent chamber is disposed above the second reagent chamber.

[0169] In another embodiment, the first piercing mechanism is configured to pierce the first pierceable seal when the first piercing mechanism moves toward the reaction chamber; and the second piercing mechanism is configured to pierce the second pierceable seal when the first piercing mechanism moves toward the reaction chamber.

[0170] In another embodiment, the first piercing mechanism is disposed above the second piercing mechanism.

[0171] In another embodiment, the first piercing mechanism is a tapered piercing mechanism with a pointed end directed toward the first pierceable seal and extending a first length.

[0172] In another embodiment, the first length of the first piercing mechanism is equal to the length of the second sealed reagent chamber extending in a direction toward the reaction chamber.

[0173] In another embodiment, the second piercing mechanism is a tapered piercing mechanism with a tip directed toward the second pierceable seal and extending a second length, and the second length of the second piercing mechanism is less than the first length of the first piercing mechanism.

[0174] In another embodiment, the tip end face of the second piercing mechanism includes a seal piercing area and a seal pressing area, wherein the seal piercing area is closer to the second pierceable seal than the seal pressing area, and along the direction toward the second pierceable seal, the length of the bottom of the seal pressing area is greater than the length of the top of the seal pressing area.

[0175] In another embodiment, the first reagent chamber includes a slidably connected first piston mechanism, the first piston mechanism is connected to the mixing device through a first piston hook, and the first piston mechanism is configured to control the lateral movement of the first reagent chamber relative to the reaction chamber.

[0176] In another embodiment, the third piercing mechanism is an arc-shaped piercing mechanism with a tip facing toward the third pierceable seal, and the top of the arc-shaped piercing mechanism has a convex tip portion.

[0177] In another embodiment, the second reagent chamber includes a slidably connected second piston mechanism, the second piston mechanism is connected to the mixing device via a second piston hook, and the second piston mechanism is configured to control the lateral movement of the second reagent chamber relative to the reaction chamber.

[0178] In another embodiment, the reaction chamber further comprises an interface for outputting materials, and the interface is arranged below the first opening.

[0179] In another embodiment, the mixing device further comprises a heater, and the heater is used to heat the reaction chamber.

[0180] The above description of the embodiments is to facilitate those of ordinary skill in the art to understand and apply the present application. It is obvious that those familiar with the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without having to pay creative labor. Therefore, the present application is not limited to the embodiments herein, and improvements and modifications made by those skilled in the art based on the contents disclosed in the present application without departing from the scope and spirit of the present application are within the scope of the present application.

Claims

1. A nucleic acid extraction device comprising a mixing device, characterized in that: The nucleic acid extraction device including the mixing device includes a sample chamber, a sample chamber cover, a filter assembly, a reaction chamber, a reaction chamber cover, and a mixing device; Wherein, along the direction of fluid flow, the sample chamber includes at least one sample chamber inlet and at least one sample chamber outlet, the filter assembly includes at least one filter assembly inlet and at least one filter assembly outlet, the reaction chamber includes at least one reaction chamber inlet and at least one reaction chamber outlet, the sample chamber cover is arranged on the at least one sample chamber inlet, and the reaction chamber cover is arranged on the at least one reaction chamber outlet; Wherein, the filter assembly can be switched between an initial state position and a transfer state position to control the fluid flow between the sample chamber and the reaction chamber. When the filter assembly is in the initial state position, the sample chamber, the filter assembly and the reaction chamber are not fluidically connected; when the filter assembly is in the transfer state position, the sample chamber, the filter assembly and the reaction chamber are fluidically connected. The sample chamber cover is provided with a hollow accommodating chamber surrounded by a fixing member and opening toward the sample chamber, a push rod extending above the sample chamber cover is slidably provided on the upper part of the hollow accommodating chamber, a movable reaction reagent chamber is provided in the middle part of the hollow accommodating chamber, and a sealing film is provided at the bottom of the hollow accommodating chamber, wherein the reaction reagent chamber is provided with a through hole, a tip is provided at the bottom of the reaction reagent chamber, and the sealing film is provided below the tip of the reaction reagent chamber; The mixing device comprises a first reagent chamber for storing reagents and a second reagent chamber for storing reagents, the first reagent chamber is in fluid communication with the reaction chamber through a first opening, and the second reagent chamber is in fluid communication with the reaction chamber through a second opening; wherein a first pierceable seal is provided at the first opening; Wherein, along the direction toward the reaction chamber, the first reagent chamber sequentially comprises a first sealed reagent chamber and a second sealed reagent chamber separated by a second pierceable seal, the first opening is arranged in the second sealed reagent chamber, and the second sealed reagent chamber comprises a first piercing mechanism and a second piercing mechanism arranged therein, the first piercing mechanism is used to pierce the first pierceable seal, and the second piercing mechanism is used to pierce the second pierceable seal; Wherein, along the direction toward the reaction chamber, the second reagent chamber comprises in sequence a third sealed reagent chamber and a sealed mixing chamber separated by a third pierceable seal, the second opening is arranged in the mixing chamber, and the mixing chamber comprises a third piercing mechanism arranged therein, and the third piercing mechanism is used to pierce the third pierceable seal; Wherein, the first sealed reagent chamber cooperates with the second piercing mechanism; Wherein, the third sealed reagent chamber cooperates with the third piercing mechanism. The radial dimension of the lower portion of the sample chamber gradually decreases, and the minimum radial dimension of the lower portion of the sample chamber is smaller than the minimum radial dimension of the reaction reagent chamber. The first piercing mechanism is arranged above the second piercing mechanism.

2. The nucleic acid extraction device comprising a mixing device according to claim 1, characterized in that: The filter assembly comprises a filter assembly body and a switch mechanism, wherein the filter assembly body comprises a pressing ring, a filter membrane and a hollow fluid channel arranged in sequence along the fluid flow direction, the inlet of the pressing ring is the at least one filter assembly inlet, and the outlet of the hollow fluid channel is the at least one filter assembly outlet; The switch mechanism is used to realize the reversible switching of the filter assembly between the initial state position and the transfer state position.

3. The nucleic acid extraction device comprising a mixing device according to claim 2, characterized in that: The hollow fluid channel is arranged obliquely, wherein an end of the fluid channel close to the sample chamber is higher than an end of the fluid channel close to the reaction chamber.

4. The nucleic acid extraction device comprising a mixing device according to claim 1, characterized in that: The first reagent chamber is arranged above the second reagent chamber, and the sample chamber is arranged above the first reagent chamber.

5. The nucleic acid extraction device comprising a mixing device according to claim 1, characterized in that: The first piercing mechanism is a pointed conical piercing mechanism with a pointed end directed toward the first pierceable seal and extending a first length.

6. The nucleic acid extraction device comprising a mixing device according to claim 5, characterized in that: The first length of the first piercing mechanism is equal to the length of the second sealed reagent chamber extending in a direction toward the reaction chamber.

7. The nucleic acid extraction device comprising a mixing device according to claim 5, characterized in that: The second piercing mechanism is a tapered piercing mechanism with a tip directed toward the second pierceable seal and extending a second length, and the second length of the second piercing mechanism is less than the first length of the first piercing mechanism.

8. The nucleic acid extraction device comprising a mixing device according to claim 7, characterized in that: The tip end face of the second piercing mechanism includes a seal piercing area and a seal pressing area, wherein the seal piercing area is closer to the second pierceable seal than the seal pressing area, and along the direction toward the second pierceable seal, the length of the bottom of the seal pressing area is greater than the length of the top of the seal pressing area.

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