A kit based on gravity-driven liquid flow and a detection method thereof

Abstract

The application discloses a kit based on gravity-driven liquid flow, comprising a box body, a liquid storage area and a reaction area arranged from top to bottom in the box body, the liquid storage area comprising a liquid flow channel and a plurality of liquid storage rods arranged on the liquid flow channel, the liquid storage rods storing liquid, the liquid storage rods being provided with liquid flow holes, the liquid storage rods being capable of sliding on the liquid storage area so that the liquid flow holes are communicated with the liquid flow channel, the reaction area comprising a reaction chamber, the reaction chamber being communicated with the liquid flow channel, when the liquid flow holes are communicated with the liquid flow channel, the liquid can flow into the reaction chamber under the action of gravity, the lower end of the reaction area being provided with a waste liquid area and an amplification area, the reaction area being provided with an isolation column at a connecting end of the waste liquid area and the amplification area, the isolation column being provided with a communication hole, the isolation column being capable of sliding on the reaction area so that the communication hole communicates the reaction area with the waste liquid area or the amplification area, so that the liquid in the reaction area flows to the waste liquid area or the amplification area under the action of gravity.

Description

Technical Field

[0001] This invention relates to the fields of medical and biological detection technology, and in particular to a reagent kit and detection method based on gravity-driven fluid flow. Background Technology

[0002] Polymerase chain reaction (PCR) can amplify the target gene in a test tube by hundreds of thousands or even millions of times. It is a highly specific, efficient, accurate, and easily automated in vitro nucleic acid amplification technique. Therefore, PCR technology has become a crucial process in gene detection and is widely used in fields such as genetic disease testing, predicting infectious virus risks, and assessing biodiversity.

[0003] Before performing PCR amplification, crude samples (such as saliva, blood, and urine samples) need to undergo a series of pretreatments, including lysis, washing, and elution, to eliminate the inhibition of DNA polymerase activity by the crude samples. The pretreatment process involves multiple operations such as adding, transferring, and discarding solutions, and also requires changing containers multiple times. The operation is cumbersome and can easily introduce secondary pollution from the external environment.

[0004] Therefore, a reagent kit and detection method based on gravity-driven fluid flow are proposed to solve the above problems. Summary of the Invention

[0005] This invention overcomes the shortcomings of the prior art and provides a reagent kit and its detection method based on gravity-driven liquid flow.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: a reagent kit and its detection method based on gravity-driven liquid flow, comprising: a box body and a liquid storage area and a reaction area arranged from top to bottom in the box body, the liquid storage area including a liquid flow channel and a plurality of liquid storage rods arranged on the liquid flow channel, the liquid storage rods storing liquid, the liquid storage rods being provided with liquid flow holes, the liquid storage rods being able to slide on the liquid storage area, so that the liquid flow holes are connected to the liquid flow channel;

[0007] The reaction zone includes a reaction chamber, which is connected to the liquid flow channel. When the liquid flow hole is connected to the liquid flow channel, the liquid can flow into the reaction chamber under the action of gravity.

[0008] The lower end of the reaction zone is provided with a waste liquid zone and an amplification zone. An isolation column is provided at the connection end between the reaction zone and the waste liquid zone and the amplification zone. The isolation column is provided with a connecting hole. The isolation column can slide on the reaction zone, so that the connecting hole connects the reaction zone with the waste liquid zone or the amplification zone, thereby allowing the liquid in the reaction zone to flow to the waste liquid zone or the amplification zone under the action of gravity.

[0009] In a preferred embodiment of the present invention, a magnet is provided on the inner wall of the reaction chamber, and a magnetic bead is placed in the reaction chamber. The magnet is capable of dragging the magnetic bead to perform reciprocating motion.

[0010] In a preferred embodiment of the present invention, when the magnetic bead reciprocates, the reaction chamber remains closed, and a heating structure is provided on the inner wall of the reaction chamber.

[0011] In a preferred embodiment of the present invention, the reaction zone, the amplification zone and the waste liquid zone are all provided with vents, and the vents can be switched to open or closed states, and remain open when the liquid is flowing.

[0012] In a preferred embodiment of the present invention, an air hole is provided on the top of the box, which, when opened, allows external air to be connected to pressurize the inside of the box.

[0013] In a preferred embodiment of the present invention, the liquid storage rod includes at least a cleaning liquid storage rod and an elution liquid storage rod, wherein the cleaning liquid storage rod contains cleaning liquid and the elution liquid storage rod contains elution liquid.

[0014] In a preferred embodiment of the present invention, there are at least two cleaning fluid reservoirs and at least one eluent reservoir, with the eluent reservoir positioned above the cleaning fluid reservoir.

[0015] In a preferred embodiment of the present invention, a sample addition port is provided on one side of the reaction chamber, through which a detection sample and magnetic beads can be added into the reaction chamber.

[0016] In a preferred embodiment of the present invention, limit plates are provided at both ends of the liquid storage rod to prevent the liquid storage rod from detaching from the liquid storage area when it slides on the liquid storage area.

[0017] In a preferred embodiment of the present invention, the isolation column is provided with a limiting block and a stop lever. The limiting block is located at one end away from the stop lever. The stop lever is provided with a first stop, a second stop, and a third stop. When the isolation column moves horizontally, its stop lever moves into the reaction zone, thereby pushing the isolation column to a designated position to achieve communication between the reaction zone and the waste liquid zone or the amplification zone.

[0018] In a preferred embodiment of the present invention, the reaction zone is provided with two chamfers to facilitate the flow of liquid into the lower channel.

[0019] In a preferred embodiment of the present invention, a tapered constriction structure is provided at the junction of the liquid storage rod and the liquid outlet, and the constriction is at an angle of 5-15° with the vertical direction.

[0020] In a preferred embodiment of the present invention, all the sealing structures, chamfers, plunger holes, plunger channels and liquid flow channels in the kit are made of hydrophobic material.

[0021] In a preferred embodiment of the present invention, a detection method for a gene detection kit based on gravity-driven fluid flow includes the following steps:

[0022] Pyrolysis: Open the sample inlet, add the sample to be tested and magnetic beads into the reaction chamber, drive the motor to drag the magnetic beads in a reciprocating motion, the analyte is adsorbed on the magnetic beads, and the waste liquid flows into the waste liquid area under the action of gravity.

[0023] Cleaning: Push the cleaning fluid reservoir rod, and the fluid flow hole connects with the fluid flow channel. The cleaning fluid flows into the reaction chamber under the action of gravity. The drive motor drags the magnetic bead to perform reciprocating motion for cleaning. After cleaning, the waste liquid flows into the waste liquid area under the action of gravity.

[0024] Sample removal: Push the eluent reservoir rod to connect the liquid flow orifice with the liquid flow channel. The eluent flows into the reaction chamber under gravity. The drive motor drags the magnetic bead in a reciprocating motion. The analyte is eluted from the magnetic bead and flows into the amplification area under gravity.

[0025] In a preferred embodiment of the present invention, the pores in the reaction zone, waste liquid zone, and amplification zone remain open when liquid flows in, thereby reducing the internal pressure in the gas discharge zone and assisting the flow of liquid in the channel.

[0026] In a preferred embodiment of the present invention, when the sample viscosity is high or other special circumstances occur, the liquid in the reagent kit is difficult to flow smoothly due to its own gravity. The air hole can be opened to change the pressure inside the reagent kit to assist the flow of liquid in the reagent kit, so as to realize the dual-driven liquid flow of gravity and pressure.

[0027] This invention addresses the shortcomings of the prior art and has the following beneficial effects:

[0028] (1) This invention arranges the storage area, reaction area, waste liquid area and amplification area from top to bottom. By pushing the plunger to reach the designated position, the liquid can flow into the designated area according to its own gravity. The liquid transfer can be completed without adding a power device. It saves energy and reduces the number of internal parts of the reagent kit, making the reagent kit lighter. Moreover, the gravity-driven method will not only not put a load on the device when detecting large-throughput liquids, but will also be more suitable. In addition, the device is also equipped with an air hole. When the sample viscosity is high or other special circumstances occur, the liquid in the reagent kit is difficult to flow smoothly according to its own gravity. The air hole can be opened to change the internal pressure of the reagent kit to assist the liquid flow in the reagent kit. It realizes the dual-driven liquid flow of gravity and pressure. The sample can be guaranteed to reach the designated position smoothly without adding any external power device. It is simple and lightweight.

[0029] (2) This invention sets up a storage area, a reaction area, a waste liquid area and an amplification area in one test kit. The liquid to be tested is cleaned and eluted by storing and releasing liquid through the storage rod in the storage area. The reaction area is separated from the waste liquid area and the amplification area by the isolation column, so as to collect the waste liquid and the amplification liquid. The operation is simple. The lysis, cleaning, elution and amplification steps can be completed in one kit without the need for solution transfer. This greatly reduces the secondary pollution of the test substance and improves the detection accuracy, while also simplifying the operation steps and improving the detection efficiency.

[0030] (3) In the reaction chamber of this invention, the magnetic beads are controlled by a magnet to rotate at high speed to complete the pyrolysis. During the pyrolysis process, the reaction chamber is in a closed state and is continuously heated, which increases the internal pressure of the reaction chamber. Under high pressure, the pyrolysis of the test substance is accelerated and the pyrolysis effect is improved. In addition, the closed state allows the magnet to move in various positions, further saving energy. At the same time, the magnetic beads can be added through the sample outlet. Users can add magnetic beads of different specifications according to the pyrolysis requirements, which improves the overall application range of the device.

[0031] (4) This invention sets up several liquid storage rods in the liquid storage area and stores liquid in the liquid storage rods as needed. The liquid storage rods are set on the liquid flow channel and have liquid flow holes. Different liquids can be released by moving different liquid storage rods to align their liquid flow holes with the liquid flow channel. When the required amount is released, the release can be stopped simply by moving the liquid storage rods to misalign their liquid flow holes with the liquid flow channel. This is simple and fast and greatly improves work efficiency. For example, it stores cleaning liquid and eluent. At the same time, the eluent storage rod is set above the cleaning liquid storage rod, and the eluent can be used to wash the pipe wall through which the cleaning liquid flows, avoiding the contamination caused by the residue of the primary cleaning liquid when other types of cleaning liquid are needed for secondary testing.

[0032] (5) This invention provides an isolation column at the connection between the reaction zone and the waste liquid zone and the amplification zone, and sets the isolation column to three positions. By pushing different positions, the following can be achieved: the reaction zone and the waste liquid zone are connected, releasing the liquid in the reaction zone into the waste liquid zone; the reaction zone and the amplification zone are connected, releasing the liquid in the reaction zone into the amplification zone; the reaction zone is isolated from the waste liquid zone and the amplification zone, and the liquid remains in the reaction zone to react. The collection of different liquids can be completed through an isolation column and a simple action, which greatly improves the work efficiency. Attached Figure Description

[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0034] Figure 1 This is a schematic diagram of the structure of the reagent kit of the present invention;

[0035] Figure 2 This is a schematic diagram of the liquid storage rod of the present invention before the liquid is released;

[0036] Figure 3 This is a schematic diagram of the liquid storage rod connecting the liquid flow channel structure of the present invention;

[0037] Figure 4 This is a schematic diagram of the initial state structure of the isolation column of the present invention;

[0038] Figure 5 This is a schematic diagram of the structure of the isolation column connecting the reaction zone and the waste liquid zone of the present invention;

[0039] Figure 6 This is a schematic diagram of the structure of the isolation column that separates the reaction zone from the waste liquid zone and the amplification zone in this invention;

[0040] Figure 7 This is a schematic diagram of the structure of the isolation column connecting the reaction region and the amplification region of the present invention.

[0041] In the diagram: 1-Reagent kit; 4-Waste liquid area; 5-Amplification area; 8-Air well;

[0042] 2-Reservoir rod; 201-Cleaning fluid reservoir rod; 202-Eluent reservoir rod; 203-Flow orifice; 204-Limiting plate; 205-Conical end;

[0043] 3-Reaction zone; 301 Sample outlet; 302 Chamfer; 303 Pores;

[0044] 6-Isolation post; 601-Limit block; 602-Gear lever; 602-First gear; 6022-Second gear; 6023-Third gear; 603-Connecting hole;

[0045] 7- Liquid flow channel; 701- Liquid flow channel below the reservoir bar; 702- Liquid flow channel above the waste liquid area; 703- Liquid flow channel above the amplification area. Detailed Implementation

[0046] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0047] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein. Therefore, the scope of protection of the invention is not limited to the specific embodiments disclosed below.

[0048] In the description of this application, it should be understood that the terms "center," "upper," "lower," "horizontal," "top," "bottom," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as limiting the scope of protection of this application. Furthermore, the terms "first," "second," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, features defined with "first," "second," etc., may explicitly or implicitly include one or more of that feature. In the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0049] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. Those skilled in the art can understand the specific meaning of these terms in this application based on the specific circumstances.

[0050] like Figure 1 As shown, the box contains a liquid storage area and a reaction area 3 arranged from top to bottom. The liquid storage area includes a liquid flow channel 701 below the liquid storage rod and several liquid storage rods 2 arranged on the liquid flow channel 701. The liquid storage rods 2 store liquid and have liquid flow holes 203. The liquid storage rods 2 can slide on the liquid storage area, so that their liquid flow holes 203 are connected to the liquid flow channel 701. The reaction area 3 includes a reaction chamber, which is connected to the liquid flow channel 701. When the liquid flow holes 203 are connected to the liquid flow channel 701, the liquid can flow into the reaction chamber under the action of gravity. The lower end of the reaction area 3 is provided with a waste liquid area 4 and an amplification area 5. The connection end between the reaction area 3 and the waste liquid area 4 and the amplification area 5 is provided with an isolation column 6. The isolation column 6 is provided with a connecting hole 603. The isolation column 6 can slide on the reaction area 3, so that its connecting hole 603 connects the reaction area 3 with the waste liquid area 4 or the amplification area 5, so that the liquid in the reaction area 3 flows to the waste liquid area 4 or the amplification area 5 under the action of gravity.

[0051] By simultaneously setting up a storage zone, reaction zone 3, waste zone 4, and amplification zone 5 within a single test kit, the liquid to be tested is cleaned and eluted by storing and releasing liquid through the storage rod 2 in the storage zone. Then, the reaction zone 3 is separated from and connected to the waste zone 4 and amplification zone 5 using the isolation column 6, thus achieving the collection of waste liquid and amplification solution. The operation is simple, and the lysis, cleaning, elution, and amplification steps can be completed within a single kit 1 without the need for solution transfer operations. This greatly reduces secondary contamination of the test substance, improves detection accuracy, simplifies the operation steps, and improves detection efficiency.

[0052] In a preferred embodiment of the present invention, a magnet is provided on the inner wall of the reaction chamber, and a magnetic bead is placed in the reaction chamber. A drive motor can drive the magnet to drag the magnetic bead in a reciprocating motion. When the magnetic bead is in a reciprocating motion, the reaction chamber remains closed. The inner wall of the reaction chamber is provided with a heating structure, which can be a heating plate with a thickness not exceeding 3 cm to reduce the volume of the reaction chamber. The movement of the magnet can be controlled by an external drive device or manual vibration to control the high-speed rotation of the magnetic bead to complete the pyrolysis. During the pyrolysis process, since the reaction chamber is closed and continuously heated, the internal pressure of the reaction chamber continuously increases. Under high pressure, the pyrolysis of the test substance is accelerated, improving the pyrolysis effect. Furthermore, the closed state can prevent the magnet from moving in various positions, further saving energy.

[0053] In a preferred embodiment of the present invention, the reaction chamber may also include a case where only lysis buffer is provided. Magnetic beads are added through the sample inlet, and a drive motor is connected outside the reaction chamber. A permanent magnet is provided on the drive motor. The drive motor drives the permanent magnet to drag the magnetic beads in reciprocating motion to achieve the lysis of the analyte.

[0054] In a preferred embodiment of the present invention, a sample addition port 301 is provided on one side of the reaction chamber, through which test samples and magnetic beads can be added into the reaction chamber. The magnetic beads can be added through the sample release port. Users can add magnetic beads of different specifications according to the pyrolysis requirements, thereby improving the overall usability of the device.

[0055] In a preferred embodiment of the present invention, air holes 303 are provided on the reaction zone 3, the amplification zone 5 and the waste liquid zone 4. The air holes 303 can be switched to open or closed. When the liquid is flowing, they remain open. An air hole 8 is provided on the top of the box. When the air hole 8 is open, it can connect to the outside air to pressurize the inside of the box.

[0056] In a preferred embodiment of the present invention, all the constriction structures 205, chamfers 302, plunger holes, plunger channels 603 and liquid flow channels in the reagent kit 1 are made of hydrophobic material to facilitate liquid flow. The reaction zone 3 is provided with two chamfers 302 to facilitate liquid flow into the lower channel.

[0057] like Figure 2-3As shown, several liquid storage rods 2 are set in the liquid storage area, and liquid is stored in the liquid storage rods 2 as needed to improve the applicability of the device. The liquid storage rods 2 are hollow plungers, which makes it easy to pull the liquid storage rods 2 out of the liquid storage area. The liquid storage rods 2 are set on the liquid flow channel 7, and liquid flow holes 203 are provided on the liquid storage rods 2. By moving different liquid storage rods 2 to align their liquid flow holes 203 with the liquid flow channel 701 set below the liquid storage rod, different liquids can be released. When the required amount is released, it is only necessary to move the liquid storage rod 2 to misalign its liquid flow holes 203 with the liquid flow channel 7 to stop the release. It is simple and fast, which greatly improves the work efficiency.

[0058] The reservoir 2 includes at least a cleaning fluid reservoir 201 and an elution fluid reservoir 202. The cleaning fluid reservoir 201 contains cleaning fluid, and the elution fluid reservoir 202 contains elution fluid. There are at least two cleaning fluid reservoirs 201, which use a circulating cleaning method to clean the waste fluid and improve the detection results. There is at least one elution fluid reservoir 202, which uses elution fluid to elute the sample to be tested adsorbed on the magnetic beads. The elution fluid reservoir 202 is located above the cleaning fluid reservoir 201, and uses elution fluid to wash the tube wall through which the cleaning fluid flows, avoiding contamination by the residue of the primary cleaning fluid when other types of cleaning fluid are needed for secondary detection.

[0059] In a preferred embodiment of the present invention, the liquid storage rod 2 may also include the case of directly filling the liquid flow channel 7. The entire liquid flow channel 7 can only be connected when the liquid flow holes 203 provided on all the liquid storage rods 2 that fill the liquid flow channel 7 are connected to the liquid flow channel 7. For example, the eluent in the eluent storage rod 202 provided at the top can only enter the reaction zone 3 through the liquid flow channel 7 when the two cleaning liquid storage rods 201 provided at the bottom are simultaneously connected to the liquid flow channel.

[0060] In a preferred embodiment of the present invention, limit plates 204 are provided at both ends of the liquid storage rod 2 to prevent the liquid storage rod 2 from detaching from the liquid storage area when it slides on the liquid storage area.

[0061] In a preferred embodiment of the present invention, a conical constriction structure 205 is provided at the connection between the liquid storage rod 2 and the liquid outlet. The constriction 205 has an angle of 5-15° with the vertical direction, which facilitates the liquid to flow out from the liquid storage rod 2.

[0062] like Figure 4-7As shown, the isolation column 6 is equipped with a limiting block 601 and a stop lever 602. The limiting block 601 is located at the end away from the stop lever 602. The stop lever 602 is equipped with a first stop 6021, a second stop 6022, and a third stop 6023. When the isolation column 6 moves horizontally, its stop lever 602 moves into the reaction zone 3, thereby pushing the isolation column 6 to a designated position to connect the reaction zone 3 with the waste liquid zone 4 or the amplification zone 5. By setting the isolation column 6 at the connection end between the reaction zone 3 and the waste liquid zone 4 and the amplification zone 5, and setting the isolation column 6 with three stops, different stops can be pushed to achieve the following: the reaction zone 3 and the waste liquid zone 4 are connected, releasing the liquid in the reaction zone 3 into the waste liquid zone 4; the reaction zone 3 and the amplification zone 5 are connected, releasing the liquid in the reaction zone 3 into the amplification zone 5; the reaction zone 3 is isolated from the waste liquid zone 4 and the amplification zone 5, and the liquid remains in the reaction zone 3 to react. The collection of different liquids can be completed through an isolation column 6 and a simple action, which greatly improves the work efficiency.

[0063] In a preferred embodiment of the present invention, a tapered constriction 205 structure is provided at the connection between the isolation column 6 and the connecting hole 603. The constriction 205 has an angle of 5-15° with the vertical direction, which facilitates the flow of liquid from the reaction chamber into the waste liquid area 4 or the amplification area 5.

[0064] A detection method for a gene detection kit 1 based on gravity-driven fluid flow includes the following steps:

[0065] Pyrolysis: Open the sample loading port 301, add the sample to be tested and magnetic beads into the reaction chamber, drive the motor to drag the magnetic beads to reciprocate, the substance to be tested is adsorbed on the magnetic beads, and the waste liquid flows into the waste liquid area 4 under the action of gravity.

[0066] Cleaning: Push the cleaning fluid storage rod 201, the liquid flow hole 203 is connected to the liquid flow channel 7, the cleaning fluid flows into the reaction chamber under the action of gravity, the drive motor drags the magnetic bead to make reciprocating motion for cleaning, after cleaning, the waste liquid flows into the waste liquid area 4 under the action of gravity;

[0067] Sample removal: Push the eluent reservoir 2, the liquid flow hole 203 is connected to the liquid flow channel 701 below the reservoir, the eluent flows into the reaction chamber under the action of gravity, the drive motor drags the magnetic bead to make reciprocating motion, the analyte is eluted from the magnetic bead and flows into the amplification area 5 under the action of gravity.

[0068] By arranging the storage area, reaction area 3, waste liquid area 4, and amplification area 5 from top to bottom, the plunger is pushed to reach the designated position, allowing the liquid to flow into the designated area according to its own gravity. Liquid transfer can be completed without the need for an additional power unit, saving energy and reducing consumption while reducing the number of internal parts of the reagent kit 1, making the reagent kit 1 more portable. Moreover, relying on gravity drive will not only not put a load on the device when detecting large-throughput liquids, but will also be more suitable, making the device unaffected by the amount of liquid to be detected and improving the applicability of the device.

[0069] In a preferred embodiment of the present invention, the vents 303 of the reaction zone 3, the waste liquid zone 4 and the amplification zone 5 remain open when liquid flows in, thereby reducing the internal pressure in the gas discharge area and assisting the flow of liquid in the channel.

[0070] In a preferred embodiment of the present invention, if the sample viscosity is high or other special circumstances occur, making it difficult for the liquid in the reagent kit 1 to flow smoothly under its own gravity, the air hole 8 can be opened to change the pressure inside the reagent kit 1, thereby assisting the flow of the liquid inside the reagent kit 1 and achieving a dual-driven liquid flow by gravity and pressure.

[0071] The specific operating procedure of this invention is as follows: The sample to be tested and the magnetic beads are added into the reaction zone 3 through the sample application port 301. The sample is lysed by the operation of the magnet dragging the magnetic beads back and forth and heating the mixture in the reaction zone through the external driving device or manual shaking of the detection box, and the nucleic acid is adsorbed on the magnetic beads. After lysis, the magnet attracts the magnetic bead, pushing the dual-channel isolation column 6 to the first position, aligning the left channel with the liquid flow channel 702 above the waste liquid zone 4, draining the liquid in the reaction zone 3 into the waste liquid zone 4. After the liquid in the reaction zone 3 is emptied, the dual-channel isolation column 6 is pushed to the second position to separate the reaction zone 3 and the waste liquid zone 4. The reservoir rod 2 containing the cleaning solution is pushed to the designated position, aligning the liquid flow orifice 203 with the liquid flow channel 701 below the reservoir rod, releasing the cleaning solution from the plunger into the reaction zone 3. The magnet continues to drive the magnetic bead to reciprocate, performing nucleic acid cleaning. After cleaning, the dual-channel isolation column 6 is pushed to the first position, aligning the left channel with the liquid flow channel 702 above the waste liquid zone 4, draining the liquid in the reaction zone 3 into the waste liquid zone 4. After the liquid is emptied, the isolation column 6 is pushed... Move to the second position to block the reaction zone 3 and the waste liquid zone 4; push the reservoir rod 2 containing the cleaning solution to the designated position and perform the same cleaning steps as above; push the reservoir rod 2 containing the elution solution to the designated position, align the liquid flow orifice 203 with the liquid flow channel 701 below the reservoir rod, release the elution solution in the plunger into the reaction zone 3, drive the magnet again to drag the magnetic bead back and forth to elute the nucleic acid adsorbed on the magnetic bead, after elution, drive the motor to control the permanent magnet to hold the magnetic bead, and at the same time push the dual-channel isolation column 6 to the third position, so that the right channel is aligned with the liquid flow channel 703 above the amplification zone 5, connecting the reaction zone 3 and the amplification zone 5, transferring the nucleic acid solution in the reaction zone 3 to the amplification zone 5, and finally heating and optical detection of the amplification zone 5 to complete the amplification and detection of the target gene.

[0072] Based on the preferred embodiments of the present invention described above, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification, but must be determined according to the scope of the claims.

Claims

1. A reagent kit based on gravity-driven fluid flow, comprising: The box body and the liquid storage area and reaction area arranged from top to bottom within the box body are characterized in that, The liquid storage area includes a liquid flow channel and a plurality of liquid storage rods disposed on the liquid flow channel. The liquid storage rods are capable of storing liquid, and liquid flow holes are provided on the liquid storage rods. The liquid storage rods are slidable on the liquid storage area so that the liquid flow holes are connected to the liquid flow channel. The reaction zone includes a reaction chamber, which is connected to the liquid flow channel. When the liquid flow hole is connected to the liquid flow channel, the liquid can flow into the reaction chamber under the action of gravity. The lower end of the reaction zone is provided with a waste liquid zone and an amplification zone. An isolation column is provided at the connection end between the reaction zone and the waste liquid zone and the amplification zone. The isolation column is provided with a connecting hole. The isolation column can slide in the reaction zone, so that the connecting hole connects the reaction zone with the waste liquid zone or the amplification zone, thereby allowing the liquid in the reaction zone to flow to the waste liquid zone or the amplification zone under the action of gravity. A magnet is provided on the inner wall of the reaction chamber, and a magnetic bead is placed in the reaction chamber. The magnet can drag the magnetic bead to make a reciprocating motion. An air vent is provided on the top of the box, which allows external air to be connected and pressurize the inside of the box when it is opened.

2. The reagent kit based on gravity-driven fluid flow according to claim 1, characterized in that: The reaction zone, amplification zone, and waste liquid zone are all provided with vents, which can be switched to open or closed, and remain open when the liquid is flowing.

3. The reagent kit based on gravity-driven fluid flow according to claim 2, characterized in that: When the magnetic bead reciprocates, the pores close, and the reaction chamber remains sealed.

4. The reagent kit based on gravity-driven fluid flow according to claim 1, characterized in that: The reservoir rod includes at least a cleaning fluid reservoir rod and an elution fluid reservoir rod, wherein the cleaning fluid reservoir rod contains cleaning fluid and the elution fluid reservoir rod contains elution fluid.

5. The reagent kit based on gravity-driven fluid flow according to claim 4, characterized in that: The cleaning fluid reservoir has at least two bars, and the eluent reservoir has at least one bar, with the eluent reservoir positioned above the cleaning fluid reservoir.

6. A reagent kit based on gravity-driven fluid flow according to claim 1, characterized in that: A sample inlet is provided on one side of the reaction chamber, through which the test sample and / or magnetic beads and / or lysis buffer can be added into the reaction chamber.

7. The detection method of the reagent kit based on gravity-driven fluid flow according to any one of claims 1-6, characterized in that, Includes the following steps: Pyrolysis: Open the sample inlet, add the sample to be tested and magnetic beads into the reaction chamber, drive the motor to drag the magnetic beads in a reciprocating motion, the analyte is adsorbed on the magnetic beads, and the waste liquid flows into the waste liquid area under the action of gravity. Cleaning: Push the cleaning fluid reservoir rod, and the fluid flow hole connects with the fluid flow channel. The cleaning fluid flows into the reaction chamber under the action of gravity. The drive motor drags the magnetic bead to perform reciprocating motion for cleaning. After cleaning, the waste liquid flows into the waste liquid area under the action of gravity. Sample removal: Push the eluent reservoir rod to connect the liquid flow orifice with the liquid flow channel. The eluent flows into the reaction chamber under gravity. The drive motor drags the magnetic bead in a reciprocating motion. The analyte is eluted from the magnetic bead and flows into the amplification area under gravity.

8. The detection method of the reagent kit based on gravity-driven fluid flow according to claim 7, characterized in that: The vents in the reaction zone, waste liquid zone, and amplification zone remain open when liquid flows in, reducing the internal pressure in the gas discharge area and assisting the liquid flow in the channel.

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