Integrated device for rapidly extracting nucleic acid through paramagnetic particle method

Abstract

The utility model relates to the field of nucleic acid detection, in particular to an integrated magnetic bead method nucleic acid rapid extraction device which comprises a tube body and a tube cover, a cavity is arranged in the tube body and comprises a magnetic bead adsorption cavity, a first washing cavity, a second washing cavity and an elution cavity which are sequentially arranged, and a first liquid channel is arranged on the inner wall of the tube body. The first isolation layer enters the first washing cavity through the first liquid channel and floats upwards to the junction of the magnetic bead adsorption cavity and the first washing cavity to separate the magnetic bead adsorption cavity from the first washing cavity; the inner wall of the pipe body is further provided with a second liquid channel, and the second isolation layer enters the second washing cavity through the second liquid channel and floats upwards to the junction of the first washing cavity and the second washing cavity to isolate the first washing cavity from the second washing cavity. The utility model ensures that the liquid position is accurate.

Description

Technical Field

[0001] The utility model relates to the field of nucleic acid detection, in particular to an integrated magnetic bead method nucleic acid rapid extraction device. Background Art

[0002] With the rapid development of genetic diagnosis, genetically modified food testing, and personalized medicine, the current nucleic acid extraction technology can no longer meet the needs of today's biotechnology, and there is an urgent need for a high-throughput, automated nucleic acid extraction method. Against this background, magnetic bead-based nucleic acid extraction came into being. Magnetic bead-based nucleic acid extraction can generally be divided into four steps: lysis-binding-washing-elution. The operation is simple and time-saving. The entire extraction process can be completed within 36-40 minutes, and the specific binding of magnetic beads to nucleic acids makes the extracted nucleic acids high in purity and concentration. In addition, since toxic reagents such as benzene and chloroform in traditional methods are not used, the harm to experimental operators is reduced to a minimum, which is in line with modern environmental protection concepts.

[0003] The applicant disclosed an integrated bottom-opening nucleic acid quick extraction test tube, quick extraction detection device and method in a Chinese utility model patent application (publication number: CN113186098A, publication date: 2021.07.30), wherein the test tube includes a main tube; the internal cavity of the main tube is provided with a lysis zone, a washing zone and an elution zone from top to bottom; the lysis zone and the washing zone and the washing zone and the elution zone are separated by a hydrophobic sealing layer, and a magnetic bead washing solution and a nucleic acid elution solution are provided in the washing zone and the elution zone respectively; the internal cavity of the main tube is also provided with a magnetic track extending from the lysis zone to the elution zone, the lower end of the magnetic track is integrally formed with the main tube and is provided with an opening connected to the outside world for the magnetic rod to extend into, and the upper end of the magnetic track is closed and located in the elution zone. When in use, the magnetic rod extends into the magnetic track, and the magnetic beads are driven by magnetic force to move from the lysis zone to the elution zone below, thereby transferring the nucleic acid cleaved out of the lysis zone and bound to the magnetic beads to the elution zone for subsequent detection.

[0004] However, the applicant found the following problems in the process of manufacturing the above-mentioned test tube: the hydrophobic sealing layer usually uses paraffin wax, and in order to set the paraffin wax at the junction of the cracking zone, the washing zone and the elution zone, it is necessary to first load the paraffin wax into the partition plug, and then load the partition plug into the inner wall of the main tube, and it is noted that there is a gap between the partition plug and the outer wall of the magnetic track for the magnetic beads to pass through, and the gap also needs to be sealed by paraffin wax, but as the partition plug moves in the test tube, the paraffin wax located on the inner side of the partition plug and the outer wall of the magnetic track are constantly in contact and friction, so that the paraffin wax is partially consumed, and the paraffin wax cannot seal the gap between the partition plug and the magnetic track, so that it cannot play a sealing role. In addition, the inner diameter of the main tube is small and there is a magnetic track in the main tube, which is not convenient for controlling the movement of the partition plug in the test tube. Utility Model Content

[0005] To solve the above problems, the purpose of the utility model is to provide an integrated magnetic bead method rapid nucleic acid extraction device, which injects corresponding liquids through the liquid channel on the tube body and forms an isolation layer to ensure that each layer of liquid flows into the correct position and separates the corresponding liquids.

[0006] For the purpose of this utility model, the following technical solutions are adopted:

[0007] An integrated magnetic bead method nucleic acid rapid extraction device comprises a tube body and a tube cover arranged on the tube body, wherein a cavity is arranged inside the tube body, and the cavity comprises a magnetic bead adsorption cavity, a first washing cavity, a second washing cavity and an elution cavity which are arranged in sequence, wherein the first washing cavity, the second washing cavity and the elution cavity are used to store a first magnetic bead washing liquid, a second magnetic bead washing liquid and a nucleic acid elution liquid respectively, wherein a first liquid channel is arranged on the inner wall of the tube body, wherein a first liquid outlet of the first liquid channel is located in the first washing cavity, wherein a density of a first isolation layer is less than that of the first magnetic bead washing liquid, wherein the first isolation layer enters the first washing cavity through the first liquid channel and floats to the junction of the magnetic bead adsorption cavity and the first washing cavity, thereby separating the magnetic bead adsorption cavity from the first washing cavity; wherein a second liquid channel is also arranged on the inner wall of the tube body, wherein a second liquid outlet of the second liquid channel is located in the second washing cavity, wherein a density of the second isolation layer is less than that of the second magnetic bead washing liquid, wherein the second isolation layer enters the second washing cavity through the second liquid channel and floats to the junction of the first washing cavity and the second washing cavity, thereby separating the first washing cavity and the second washing cavity.

[0008] Preferably, a third isolation layer is provided between the second washing chamber and the elution chamber to separate the two.

[0009] Preferably, the first isolation layer, the second isolation layer and the third isolation layer are made of one of paraffin, p-bromoanisole, 1-bromo-3-chloropropane and vaseline.

[0010] Preferably, a magnetic rod channel extending between the magnetic bead adsorption chamber, the first washing chamber and the second washing chamber is also provided in the tube body, the lower end of the magnetic rod channel is at least located at the bottom position of the third isolation layer, and a magnetic rod entrance for the magnetic rod to enter is provided at one end of the magnetic rod channel.

[0011] Preferably, the tube body comprises a base, on which is provided a connecting rod extending upward, the connecting rod is hollow inside and has an opening at the lower end, the opening being the magnetic rod entrance, and the magnetic rod channel is formed inside the connecting rod.

[0012] Preferably, the base is also provided with a connection port opening upward, the second washing chamber is located in the connection port, a plurality of independent reaction tubes are provided at the lower part of the base, a collection tank is provided in the reaction tubes, the elution chamber is located in the collection tank, and the third isolation layer is located at the slot position of the collection tank.

[0013] Preferably, the reaction tubes are evenly distributed along the circumferential direction, and the notch of the collecting tank is in a funnel shape with a larger top and a smaller bottom;

[0014] A second partition is provided on the connection port, the second partition covers the upper part of the connection port, and a liquid injection port is left at the second liquid outlet;

[0015] A first partition is arranged inside the pipe body, and the first partition is connected with the second partition to form a second partition plug, and the second isolation layer is located inside the second partition plug.

[0016] Preferably, a first partition plug and a second partition plug are provided in the tube body, the first partition plug corresponds to the position of the first liquid outlet, the first isolation layer is formed inside the first partition plug, and an exhaust hole is provided on the top of the first partition plug; the second partition plug corresponds to the position of the second liquid outlet, the second isolation layer is formed inside the second partition plug, and an exhaust hole is provided on the top of the second partition plug.

[0017] Preferably, the first liquid inlet of the first liquid channel is located at the top of the tube body, the second liquid inlet of the second liquid channel is located at the top of the tube body, and the tube cover seals the first liquid inlet and the second liquid inlet;

[0018] The tube cover has a storage part, a storage opening is formed at the lower end of the storage part, a bottom seal is arranged at the lower part of the storage opening, and a closed space is formed inside the storage opening; the tube cover is connected to the tube body by a thread, and a puncture part is arranged at the top of the magnetic rod channel, which is arranged toward the bottom seal and can pierce the bottom seal.

[0019] Preferably, a clamping portion is provided on the side wall of the device; the clamping portion is a first clamping block provided on the side wall of the tube body and protruding outward, or the clamping portion is a second clamping block provided on the side wall of the base and protruding outward.

[0020] To sum up, the beneficial effects of the utility model are as follows: a first liquid channel and a second liquid channel are set on the inner wall of the tube body, and the corresponding liquid and the isolation layer are injected into the tube body through the liquid channels. There is no need to use a separation plug to load paraffin into the test tube as in the prior art, which simplifies the liquid injection process and ensures that the position of the liquid is accurate, thereby playing a separation role. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a three-dimensional diagram of the integrated magnetic bead-based rapid nucleic acid extraction device.

[0022] Figure 2 This is a disassembled diagram of the integrated magnetic bead-based rapid nucleic acid extraction device.

[0023] Figure 3 for Figure 1 Cross-sectional view at AA in the middle.

[0024] Figure 4 for Figure 1 Cross-sectional view at the middle BB

[0025] Figure 5 A cross-sectional view of the tube body.

[0026] Figure 6 It is a three-dimensional diagram of the first partition plug.

[0027] Figure 7 A three-dimensional diagram of the base.

[0028] Figure 8 A schematic diagram of the interior of the base.

[0029] Fig. 9 A schematic diagram of the lower part of the base. DETAILED DESCRIPTION

[0030] like Figure 1 and 2 As shown, a rapid nucleic acid extraction device is particularly suitable for magnetic bead method nucleic acid extraction. The device is in the shape of a test tube, including: a tube body 200 and a tube cover 100 arranged on the top of the tube body 200. The tube cover 100 is usually designed to be detachable or partially open, so as to add the sample to be tested and the reagent required for the test into the tube body 200. The tube body 200 includes a main tube and a base 300 arranged at the lower end of the main tube. In this embodiment, the base 300 is detachably arranged on the main tube, and the nucleic acid material extracted below is located in the base 300, so that after the extraction is completed, the base 300 can be removed from the main tube and taken to other places for nucleic acid testing.

[0031] like Figure 2 and 3As shown, the tube body 200 is a hollow tube with an upper opening, and a longitudinally extending cavity is arranged inside the tube body 200. The cavity is divided into: a magnetic bead adsorption cavity 201, a washing cavity 202 and an elution cavity 203 from top to bottom, and a magnetic bead washing liquid (not shown) and a nucleic acid elution liquid (not shown) are arranged in the washing cavity 202 and the elution cavity 203 respectively. It should be noted that the magnetic bead washing liquid and the nucleic acid elution liquid are usually arranged in the test tube when the test tube is assembled, that is, before the nucleic acid is extracted, the magnetic bead washing liquid and the nucleic acid elution liquid have been arranged in the test tube in advance, rather than temporarily loaded. The reagents (such as nucleic acid lysis solution, samples and magnetic beads) in the magnetic bead adsorption chamber 201 can be temporarily loaded, for example, when the test tube is assembled, the lysis solution (not shown) or the mixture of lysis solution and magnetic beads is added to the magnetic bead adsorption chamber 201; when nucleic acid extraction is required, the lysis solution or the mixture of lysis solution and magnetic beads can be temporarily added to the magnetic bead adsorption chamber 201; when nucleic acid extraction is required, the lysis solution can be directly placed in the magnetic bead adsorption chamber 201 together with the magnetic beads. It should also be noted that in this embodiment, the elution chamber 203 is located in the base 300, and the nucleic acid elution solution and the third isolation layer described below are separately injected after the base 300 is disassembled from the tube body 200.

[0032] In order to avoid direct contact between the three liquids in the magnetic bead adsorption chamber 201, the washing chamber 202 and the elution chamber 203, a first isolation layer 210 is provided between the magnetic bead adsorption chamber 201 and the washing chamber 202 to separate the two, and a third isolation layer (not shown) is provided between the washing chamber 202 and the elution chamber 203 to separate the two, that is, the lysate and the magnetic bead washing solution and the magnetic bead washing solution and the nucleic acid elution solution are separated by the first isolation layer 210 and the third isolation layer respectively. It should be noted that since the lysate, the magnetic bead washing solution and the nucleic acid elution solution are all water-soluble liquids, in order to ensure the isolation effect, the first isolation layer 210 and the third isolation layer are both hydrophobic layers, such as paraffin, a hydrophobic liquid layer, and specifically can be one or more of p-bromoanisole, 1-bromo-3-chloropropane, and vaseline.

[0033] Further, if Figure 3As shown, in order to clean the impurities on the surface of the magnetic beads, the washing chamber 202 is divided into a first washing chamber 2021 and a second washing chamber 2022. The first washing chamber 2021 is located above the second washing chamber 2022, that is, the first washing chamber 2021 is adjacent to the magnetic bead adsorption chamber 201, and the second washing chamber 2022 is adjacent to the elution chamber 203. The first washing chamber 2021 and the second washing chamber 2022 are separated by a second isolation layer 220. Similar to the first isolation layer 210 and the third isolation layer, the second isolation layer 220 is also a hydrophobic layer, such as paraffin, a hydrophobic liquid layer, etc. The first washing chamber 2021 and the second washing chamber 2022 are respectively provided with a first magnetic bead washing liquid (not shown) and a second magnetic bead washing liquid (not shown). The first magnetic bead washing liquid is used to remove impurities such as proteins in nucleic acids combined with magnetic beads, and the second magnetic bead washing liquid is used to remove impurities such as inorganic salts brought down from the first magnetic bead washing liquid.

[0034] like Figure 3 As shown, a magnetic bar channel 230 extending between the magnetic bead adsorption chamber 201, the first washing chamber 2021 and the second washing chamber 2022 is provided at the central position of the inner cavity of the tube body 200, the lower end of the magnetic bar channel 230 is at least located at the bottom position of the third isolation layer, and a magnetic bar entrance 231 for a magnetic bar (not shown) to enter is formed at one end of the magnetic bar channel 230, that is, after the magnetic bar extends into the magnetic bar channel 230, it can move between the magnetic bead adsorption chamber 201, the first washing chamber 2021 and the second washing chamber 2022, thereby driving the magnetic beads (not shown) to move accordingly in the test tube, and can at least move to the position at the bottom of the third isolation layer, so that the magnetic beads enter the elution chamber 203. Preferably, the lower end of the magnetic bar channel 230 extends to the elution chamber 203, so that the magnetic bar can drive the magnetic beads to move directly into the nucleic acid elution solution.

[0035] When the test tube is used, the tube cover 100 is opened, and the sample of nucleic acid to be extracted, a predetermined amount of nanomagnetic beads and a lysis solution are placed in the magnetic bead adsorption chamber 201. The lysis solution lyses the sample so that the nucleic acid substance in the sample is dissolved in the lysis solution and combined with the magnetic beads. The test tube can be rotated and vibrated during the process to accelerate the lysis process. After sufficient reaction, the magnetic bar is inserted from the magnetic bar entrance 231 and moved into the magnetic bead adsorption chamber 201. At this time, the magnetic beads are adsorbed on the outer wall of the magnetic bead channel 230 under the action of magnetic force, and then the magnetic bar is moved downward. The magnetic beads follow the magnetic bar and move synchronously downward along the outer wall of the magnetic bead channel 230, so that the magnetic beads and the nucleic acid substances thereon pass through the first isolation layer 210 and enter the first washing chamber 2021 downward (if the first isolation layer 210, the second isolation layer 220 and the third isolation layer are paraffin, the test tube needs to be heated in advance to melt the paraffin). In the first washing chamber 2021, the first magnetic bead washing solution washes the surface of the magnetic beads to remove impurities such as proteins in the nucleic acid bound to the magnetic beads, and then the magnetic bar continues to move downward, and the magnetic beads pass through the second isolation layer 220 to enter the second washing chamber 2022. In the second washing chamber 2022, the second magnetic bead washing solution washes the surface of the magnetic beads to remove impurities such as inorganic salts brought down from the first magnetic bead washing solution. Afterwards, the magnetic beads continue to move downward following the magnetic bar through the third isolation layer into the elution chamber 203, and the nucleic acid substances bound to the magnetic beads are transferred to the nucleic acid elution solution. During detection, PCR or isothermal amplification reactions can be performed directly in a test tube, and dyes, probes, primers, enzymes, and buffers can be introduced into the nucleic acid elution solution, so that the external device can observe the PCR or isothermal amplification reaction in real time through fluorescence quantitative detection, and the number of sample nucleic acids can be obtained by analyzing the fluorescence value; or the base 300 is unplugged from the tube body 200 and taken to other devices for nucleic acid detection.

[0036] In order to load the above-mentioned first isolation layer 210, second isolation layer 220 and first magnetic bead washing solution and second magnetic bead washing solution into the tube body 200, a first liquid channel 240 and a second liquid channel 250 are provided on the inner wall of the tube body 200. The first liquid channel 240 is used to inject the first magnetic bead washing solution and the first isolation layer 210 into the first washing chamber 2021 in sequence, and the second liquid channel 250 is used to inject the second magnetic bead washing solution and the second isolation layer into the second washing chamber 2022 in sequence.

[0037] Specifically, Figure 4 and 5As shown, the first liquid inlet 241 of the first liquid channel 240 is located at the top of the tube body 200, the first liquid outlet 242 of the first liquid channel 240 is located in the first washing chamber 2021, the second liquid inlet 251 of the second liquid channel 250 is located at the top of the tube body 200, and the second liquid outlet 252 of the second liquid channel 250 is located in the second washing chamber 2022. It can be seen that the first liquid outlet 242 is located above the second liquid outlet 252, and is located in the first washing chamber 2021 and the second washing chamber 2022 respectively, so that the liquid in the corresponding liquid channel will flow into the corresponding position. In addition, it should be noted that the density of the first isolation layer 210 is less than that of the first magnetic bead washing liquid, and the density of the second isolation layer 220 is less than that of the second magnetic bead washing liquid, so that when the liquid is injected, the first isolation layer 210 floats above the first magnetic bead washing liquid, and the second isolation layer 220 floats above the second magnetic bead washing liquid.

[0038] In this embodiment, the materials of the first isolation layer 210, the second isolation layer 220 and the third isolation layer are all paraffin wax, and how to inject these liquids into the tube body 200 is explained. First, the base 300 is separated from the main tube, and then the nucleic acid eluent is injected into the base 300 alone, and then the paraffin wax is added. After the paraffin wax is cooled, the third isolation layer mentioned above is formed, and then the base 300 is installed to the lower end of the main tube. Then a predetermined amount of the second magnetic bead washing liquid is injected from the second liquid inlet 251, so that the second magnetic bead washing liquid flows out from the second liquid outlet 252. Since a solid third isolation layer has been formed below the second washing chamber 2022, the second magnetic bead washing liquid will be kept above the third isolation layer. Then, a predetermined amount of liquid paraffin is injected from the second liquid inlet 241, so that the paraffin enters the second washing chamber 2022 from the second liquid outlet 252. Note that the density of paraffin is less than that of the second magnetic bead washing liquid, so the paraffin will float to the top of the second magnetic bead washing liquid, thereby forming a second isolation layer 220 above the second magnetic bead washing liquid, and wait for the paraffin to cool down, and the second isolation layer 220 becomes solid. After that, a predetermined amount of magnetic bead washing liquid is injected from the first liquid inlet 241, so that the magnetic bead washing liquid enters the first washing chamber 2021 from the first liquid outlet 242. Since the paraffin below has solidified, the magnetic bead washing liquid will be kept above the second isolation layer 220. Then, a predetermined amount of liquid paraffin is injected from the first liquid inlet 241, so that the paraffin enters the first washing chamber 2021 from the second liquid outlet 252. Similarly, since the density of paraffin is less than that of the magnetic bead washing liquid, the paraffin will float to the top of the magnetic bead washing liquid, thereby forming a first isolation layer 210 above the magnetic bead washing liquid, and the paraffin is cooled and the first isolation layer 210 becomes solid. If necessary, the lysis solution can be directly injected from the opening at the top of the tube body 200.

[0039] When the isolation layer adopts a hydrophobic liquid layer, the method is the same as above, but it should be noted that the hydrophobic liquid layer will not solidify like paraffin. Therefore, in order to ensure that the third isolation layer can separate the second magnetic bead washing solution and the nucleic acid elution solution, the density of the third isolation layer should be between the second magnetic bead washing solution and the nucleic acid elution solution, and to ensure that the density of the second isolation layer 220 should be between the first magnetic bead washing solution and the second magnetic bead washing solution.

[0040] According to the above method, the first isolation layer 210 enters the first washing chamber 2021 through the first liquid channel 240 and floats to the junction of the magnetic bead adsorption chamber 201 and the first washing chamber 2021. The second isolation layer 220 enters the second washing chamber 2022 through the second liquid channel 250 and floats to the junction of the first washing chamber 2021 and the second washing chamber 2022.

[0041] Through the above method, the isolation layer is no longer formed by using a separation plug as in the prior art. The structure of the present application simplifies the liquid injection method, which is conducive to the formation of the first isolation layer 210, the second isolation layer 220 and the third isolation layer, and does not cause the problem of the isolation layer failing to perform a sealing function in the prior art.

[0042] Further, in this embodiment, the first liquid channel 240 is implemented in the following manner: Figure 5 As shown, a first extension wall 243 is integrally formed on the inner wall of the left side of the tube body 200. The first extension wall 243 is semicircular. The two connecting ends 244 ( Figure 5 is a cross-sectional view, showing only one of the connection ends 244) disposed on the inner wall of the pipe body 200, and the first extension wall 243 extends toward the inner side of the pipe body 200 ( Figure 5 The first extension wall 243 protrudes on the right side of the middle portion, so that a first longitudinally extending channel is formed between the inner wall of the first extension wall 243 and the inner wall of the tube body 200, and the upper end of the first channel is located at the top of the tube body 200, and the lower end of the first channel is located at the first washing chamber 2021, and the first channel is the first liquid channel 240.

[0043] In this embodiment, the second liquid channel 250 is implemented in the following manner: Figure 5 As shown, a second extension wall 253 is integrally formed on the inner wall of the right side of the tube body 200. The second extension wall 253 is semicircular. Two connecting ends 254 of the second extension wall 253 are arranged on the inner wall of the tube body 200, and the second extension wall 253 extends to the inner side of the tube body 200 ( Figure 5 The second extension wall 253 protrudes on the left side of the middle part, so that a second longitudinally extending channel is formed between the inner wall of the second extension wall 253 and the inner wall of the tube body 200, and the upper end of the second channel is located at the top of the tube body 200, and the lower end of the second channel is located at the second washing chamber 2022, and the second channel is the second liquid channel 250.

[0044] like Figure 5 As shown, the first liquid channel 240 and the second liquid channel 250 are arranged on opposite sides of the inner wall of the tube body 200 to avoid stress concentration. At the same time, since the distance between the two is relatively large, it is also convenient for liquid injection.

[0045] It should be noted that when the material of the first isolation layer 210 and the second isolation layer 220 is paraffin, a separation plug can still be provided in the tube body 200 as in the prior art, for example, Figure 3 , 4 In Figures 5 and 6, a first partition plug 260 and a second partition plug 270 are provided in the inner cavity of the tube body 200, the first isolation layer 210 is located inside the first partition plug 260, and the second isolation layer 220 is located inside the second partition plug 270. However, the function of the partition plug in this embodiment is no longer to load the paraffin into the test tube, but to facilitate the solidification and molding of the paraffin.

[0046] Figure 5 and Figure 6 The schematic diagram of the first partition plug 260 is shown. The shape of the first partition plug 260 is roughly adapted to the inner cavity of the tube body 200. The first notch 261 and the second notch 262 adapted to the shape of the first extension wall 243 and the second extension wall 253 are respectively arranged on both sides of the first partition plug 260. The center of the first partition plug 260 is provided with a center hole 263 adapted to the outer wall of the magnetic bar channel 230. It is worth noting that a plurality of convex ribs 264 are evenly distributed along the circumference on the inner wall of the center hole 263. Usually, the convex ribs 264 are longitudinally extended strips. Due to the existence of the convex ribs 264, the first partition plug 260 can be matched on the magnetic bar channel 230 without moving randomly in the tube body 200. In addition, due to the provision of the convex ribs 264, a gap for the magnetic beads to pass through is formed between the inner wall of the first partition plug 260 and the outer wall of the magnetic bar channel 230. In addition, the first partition plug 260 is a hollow structure with a chamber formed inside, and a plurality of through holes 265 are arranged on the peripheral wall of the first partition plug 260, and these through holes 265 are connected to the internal chamber, and the first liquid outlet 242 is located at the position of the through holes 265, so that after the paraffin flows out from the first liquid outlet 242, it enters the chamber from these through holes 265, and the paraffin is kept in the chamber by means of the first partition plug 260, which helps the solidification of the paraffin. More importantly, the top surface of the first partition plug 260 is provided with an exhaust hole 266, so that when the paraffin solidifies, the internal gas is discharged from the exhaust hole 266, so that the paraffin can cover the entire inner cavity of the tube body 200 without leaving any gaps, and as the paraffin gradually cools from the outside to the inside, a dense first isolation layer 210 is formed in the tube body 200, ensuring that the liquid below is separated from the liquid above.

[0047] In addition, a plurality of upwardly protruding stirring rods 267 are provided on the top surface of the first partition plug 260. The stirring rods 267 are distributed around the circumferential direction. The stirring rods 267 are usually integrally formed on the first partition plug 260. During the lysis process, the test tube is shaken or vibrated, and the sample and the lysis solution collide with the stirring rods 267, which have a stirring effect to accelerate the reaction process.

[0048] In order to ensure that the first partition plug 260 can be accurately positioned in the tube body 200 , a step surface (not shown) is provided on the inner wall of the tube body 200 , and the lower end of the first partition plug 260 fits on the step surface, thereby achieving the positioning of the first partition plug 260 .

[0049] like Figure 4 and 5 As shown, the shape of the second partition plug 270 is substantially the same as that of the first partition plug 260, but preferably, the second partition plug 270 includes a first partition portion 271 and a second partition portion 272 which can be separated from each other, the first partition portion 271 is integrally formed on the inner wall of the tube body 200, and the second partition portion 272 is located on the base 300 (see Figure 7 ), the second partition 272 is in the shape of a disc. The second partition 272 serves as the bottom of the second partition plug 270 and can also serve as a cover of the base 300 to ensure that after the base 300 is removed from the tube body 200, the liquid inside the base 300 will not leak out.

[0050] A chamber is formed between the first partition 271 and the second partition 272 as in the first partition plug 260, and a plurality of through holes are also provided in the circumferential direction of the second partition plug 270, and the position of the second liquid outlet 252 corresponds to the through hole, so that after the paraffin flows out of the second liquid outlet 252, it flows into the interior of the second partition plug 270; in addition, a gap is also left between the second partition plug 270 and the outer wall of the magnetic rod channel 230 for the magnetic beads to pass through, and an exhaust hole is also provided on the top surface of the second partition plug 270, and the function of the exhaust hole is the same as that in the first partition plug 260.

[0051] like Figure 3 and 4 As shown, in this embodiment, the upper end of the magnetic bar channel 230 is closed, and the magnetic bar entrance 231 is located at the bottom of the base 300. It can also be considered that the magnetic bar entrance 231 is located at the bottom of the tube body 200 or the test tube.

[0052] As a specific implementation of the magnetic bar channel 230, in this embodiment, as Figure 7 As shown, a connecting rod 310 extending longitudinally upward and in a columnar shape is integrally formed on the base 300 . The connecting rod 310 has a closed upper end, a hollow interior and an open lower end. The opening is the magnetic rod inlet 231 , and the interior of the connecting rod 310 is the magnetic rod channel 230 .

[0053] like Figure 8 As shown, the base 300 is also provided with a connection port 301 opening toward the upper part, the connection port 301 is connected to the inside of the tube body 200, the second washing chamber 2022 is formed in the connection port 301, and the second partition 272 mentioned above is provided above the connection port 301 (see Figure 7 ), the second partition 272 closes the upper part of the connecting port 301, but a liquid injection port 302 for liquid to enter is formed at the position of the second liquid channel 250, so that the liquid flowing out of the second liquid outlet 252 can flow into the connecting port 301.

[0054] like Figure 7 As shown, in order to install the second partition 272 on the connecting port 301, upwardly protruding buckles 320 are provided on opposite sides of the base 300, the inner side surface of the buckle 320 is an inclined surface, and a snap-fit ​​groove is formed at the lower end of the buckle 320, and the second partition 272 is located in the snap-fit ​​groove, so that the second partition 272 is retained on the buckle 320.

[0055] like Fig. 9 As shown, the lower part of the base 300 is also provided with a plurality of mutually independent reaction tubes 330. Usually, the number of the reaction tubes 330 is 2-8. The reaction tubes 330 have a collection tank 303 open upward (see Figure 3 ), when the nucleic acid eluent is injected, the nucleic acid eluent flows into the collecting tank 303 and finally enters the interior of the reaction tube 300. The elution chamber 203 mentioned above is located in the reaction tube 330. The third isolation layer mentioned above is located at the notch 3031 of the collecting tank 303 to encapsulate the eluent inside the reaction tube 330. And it is noted that the diameter of the notch 3031 is small. Even if the density of paraffin is greater than that of the nucleic acid eluent, under the action of the surface tension of the liquid, the paraffin will still remain at the position of the notch 3031 and will not sink below the nucleic acid eluent. The purpose of setting up multiple reaction tubes 330 is to form multiple control groups when performing nucleic acid detection.

[0056] In addition, in order to facilitate the magnetic beads to evenly enter each reaction tube 330 when moving downward, the reaction tubes 330 are evenly distributed around the circumference, and the notches 3031 are funnel-shaped. Specifically, Figure 8 As shown, the slot 3031 includes an inclined connecting surface 3032, and the top ends of the connecting surfaces 3032 of two adjacent slots 3031 intersect to form a connecting line 3033. The connecting line 3033 extends radially so that each reaction tube 330 has the same area in the circumferential direction, so that the magnetic beads will evenly enter each slot 3031 during the process of moving downward along the outer wall of the magnetic bar channel 230, and finally be evenly distributed in each reaction tube 330.

[0057] In order to facilitate the installation and fixation of the base 300 and prevent the base 300 from being installed upside down, a first connection part is provided on one side of the base 300, and a second connection part is provided on one side of the tube body 200, and the first connection part is matched in the second connection part.

[0058] Specifically, Figure 2 As shown, the first connecting part is a groove 340 fixedly provided on the base 300, and the second connecting part is a protrusion 280 provided on the side of the tube body 200 and protruding downward. The protrusion 280 is inserted into the groove 340 and applies force to the position of the groove 340 to fix the base 300 on the tube body 200.

[0059] Furthermore, the base 300 is provided with an annular flange 350 protruding radially outward, and the annular flange 350 abuts against the bottom of the main pipe to serve as a limit when connected with the main pipe. The groove 340 is provided on the annular flange 350 .

[0060] like Figure 4 As shown, in order to prevent the liquid in the test tube from leaking out from below, a sealing ring 600 is still provided between the base 300 and the inner wall of the main tube.

[0061] like Figure 4 As shown, the tube cover 100 is disposed on the top of the tube body 200 , and the tube cover 100 closes the first liquid inlet 241 and the second liquid inlet 251 to prevent the magnetic bead washing solution from flowing out of the tube body 200 .

[0062] Furthermore, the tube cover 100 has a storage portion 110, which is cylindrical and extends downward into the tube body 200. A storage port 111 is formed at the lower end of the storage portion 110, and a bottom seal (not shown) is provided at the lower portion of the storage port 111, thereby forming a relatively closed space in the storage port 111, in which magnetic beads can be placed, so that the magnetic beads are stored in the test tube and no additional container is needed to carry them.

[0063] The material of the bottom cover can be tin foil or other materials that are easy to tear or puncture. In order to facilitate puncturing the tin foil, the tube cover 100 and the tube body 200 can be connected by threads, and the top of the magnetic bar channel 230 is provided with a puncture portion 232 that is arranged toward the bottom cover and can puncture the bottom cover. When in use, the tube cover 100 is screwed downward to move the tin foil downward and eventually punctured by the puncture portion 232, and then the tube cover 100 is screwed in the reverse direction to remove the puncture portion 232 from the tin foil, so that the magnetic beads in the tube cover 100 will fall into the magnetic bead adsorption chamber 201.

[0064] When the test tube is used for nucleic acid testing, it is usually necessary to rotate. In order to facilitate the driving device to drive the test tube to rotate, a clamping part is provided on the test tube to facilitate connection with the driving device. Specifically, Figure 1 and2 As shown, a first snap-fit ​​block 290 protruding outward is provided on the side wall of the tube body 200. When in use, the first snap-fit ​​block 290 is snapped into the groove of the driving device, thereby locking the test tube circumferentially, so that the driving device can drive the test tube to rotate. In addition, since the base 300 may be removed separately for testing, a second snap-fit ​​block 360 protruding outward is provided on the side wall of the base 300, so that when the base 300 is removed, the driving device can drive the base 300 to rotate separately.

[0065] In summary, the beneficial effects of the utility model are as follows: a first liquid channel 240 and a second liquid channel 250 are provided on the inner wall of the tube body 200, and the corresponding liquid and the isolation layer are injected into the tube body 200 through the liquid channels. There is no need to use a separation plug to load paraffin into the test tube as in the prior art, which simplifies the liquid injection process and ensures that the position of the liquid is accurate, thereby playing a separation role.

[0066] The above is a description of the embodiments of the utility model. Through the above description of the disclosed embodiments, professionals and technicians in the field can implement or use the utility model. Various modifications to these embodiments will be obvious to professionals and technicians in the field. The general principles defined in this article can be implemented in other embodiments without departing from the spirit or scope of the utility model. Therefore, the utility model will not be limited to these embodiments shown in this article, but will conform to the widest range consistent with the principles and novelties disclosed in this article.

Claims

1. An integrated magnetic bead method nucleic acid rapid extraction device, comprising a tube body (200) and a tube cover (100) arranged on the tube body (200), wherein a cavity is arranged inside the tube body (200), and the cavity comprises a magnetic bead adsorption chamber (201), a first washing chamber (2021), a second washing chamber (2022) and an elution chamber (203) arranged in sequence, wherein the first washing chamber (2021), the second washing chamber (2022) and the elution chamber (203) are used to store a first magnetic bead washing solution, a second magnetic bead washing solution and a nucleic acid elution solution, respectively, and characterized in that: A first liquid channel (240) is provided on the inner wall of the tube body (200); a first liquid outlet (242) of the first liquid channel (240) is located in the first washing chamber (2021); the density of the first isolation layer (210) is lower than that of the first magnetic bead washing liquid; the first isolation layer (210) enters the first washing chamber (2021) through the first liquid channel (240) and floats to the junction of the magnetic bead adsorption chamber (201) and the first washing chamber (2021), thereby separating the magnetic bead adsorption chamber (201) and the first washing chamber (2021); A second liquid channel (250) is also provided on the inner wall of the tube body (200); a second liquid outlet (252) of the second liquid channel (250) is located in the second washing chamber (2022); the density of the second isolation layer (220) is lower than that of the second magnetic bead washing liquid; the second isolation layer (220) enters the second washing chamber (2022) through the second liquid channel (250) and floats to the junction of the first washing chamber (2021) and the second washing chamber (2022), thereby separating the first washing chamber (2021) from the second washing chamber (2022).

2. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 1, characterized in that: A third isolation layer is provided between the second washing chamber (2022) and the elution chamber (203) to separate the two.

3. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 2, characterized in that: The components of the first isolation layer (210), the second isolation layer (220) and the third isolation layer are one of paraffin, p-bromoanisole, 1-bromo-3-chloropropane and vaseline.

4. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 2, characterized in that: A magnetic bar channel (230) extending between the magnetic bead adsorption chamber (201), the first washing chamber (2021) and the second washing chamber (2022) is also provided in the tube body (200); the lower end of the magnetic bar channel (230) is at least located at the bottom of the third isolation layer; and a magnetic bar inlet (231) for the magnetic bar to enter is provided at one end of the magnetic bar channel (230).

5. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 4, characterized in that: The tube body (200) comprises a base (300), on which a connecting rod (310) extending upward is arranged, the connecting rod (310) is hollow inside and has an opening at the lower end, the opening being the magnetic rod entrance (231), and the magnetic rod channel (230) is formed inside the connecting rod (310).

6. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 5, characterized in that: The base (300) is also provided with a connection port (301) opening upwards, the second washing chamber (2022) is located in the connection port (301), a plurality of mutually independent reaction tubes (330) are provided at the lower part of the base (300), a collection tank (303) is provided in the reaction tube (330), the elution chamber (203) is located in the collection tank (303), and the third isolation layer is located at the notch (3031) of the collection tank (303).

7. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 6, characterized in that: The reaction tubes (330) are evenly distributed along the circumferential direction, and the notch (3031) of the collecting groove (303) is in a funnel shape with a larger top and a smaller bottom; A second partition (272) is provided on the connection port (301), the second partition (272) covers the upper part of the connection port (301), and a liquid injection port (302) is left at the second liquid outlet (252); A first partition (271) is provided inside the tube body (200), and the first partition (271) and the second partition (272) are connected to form a second partition plug (270), and the second isolation layer (220) is located inside the second partition plug (270).

8. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 1, characterized in that: A first partition plug (260) and a second partition plug (270) are provided in the tube body (200); the first partition plug (260) corresponds to the position of the first liquid outlet (242); the first isolation layer (210) is formed inside the first partition plug (260); and an exhaust hole is provided at the top of the first partition plug (260); the second partition plug (270) corresponds to the position of the second liquid outlet (252); the second isolation layer (220) is formed inside the second partition plug (270); and an exhaust hole is provided at the top of the second partition plug (270).

9. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 1, characterized in that: The first liquid inlet (241) of the first liquid channel (240) is located at the top of the tube body (200), the second liquid inlet (251) of the second liquid channel (250) is located at the top of the tube body (200), and the tube cover (100) seals the first liquid inlet (241) and the second liquid inlet (251); The tube cover (100) comprises a storage portion (110), a storage opening (111) is formed at the lower end of the storage portion (110), a bottom seal is provided at the lower portion of the storage opening (111), and a closed space is formed in the storage opening (111); the tube cover (100) and the tube body (200) are connected via threads, and a piercing portion (232) is provided at the top of the magnetic bar channel (230) and is arranged towards the bottom seal and capable of piercing the bottom seal.

10. The integrated magnetic bead method nucleic acid rapid extraction device according to claim 1, characterized in that: A clamping portion is provided on the side wall of the device; the clamping portion is a first clamping block (290) provided on the side wall of the tube body (200) and protruding outwards, or the clamping portion is a second clamping block (360) provided on the side wall of the base (300) and protruding outwards.

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