Nano-magnetic beads control test tubes and gene detection reagent packaging and gene detection methods

Abstract

This invention discloses a nano-magnetic bead-controlled test tube and a method for packaging gene detection reagents and performing gene detection. The test tube includes a sample processing chamber, a washing chamber, and an amplification chamber. A sample isolation chamber and a capillary channel are provided between the sample processing chamber and the washing chamber, and an amplification isolation chamber and a capillary channel are provided between the washing chamber and the amplification chamber. A high-viscosity isolation solution is injected into the isolation chamber, forming a stable liquid isolation layer within the capillary channel. During packaging, the primer and probe solutions and the reaction system in the amplification chamber are sealed with a hydrophobic material. The isolation solution is injected into the isolation chamber, and the sample processing chamber contains the sample processing system and nano-magnetic beads. During detection, the sample releases nucleic acid in the sample processing chamber, which is adsorbed by the magnetic beads. Guided by an external magnetic field, the magnetic beads sequentially pass through the capillary channel of the sample isolation chamber into the washing chamber for washing, and then through the capillary channel of the amplification isolation chamber into the amplification chamber. After heating and amplification, fluorescence detection is performed. This invention achieves stable liquid isolation between the chambers, resulting in high detection accuracy and repeatability.

Description

Technical Field

[0001] This invention relates to the field of molecular point detection technology, and more specifically to a nanomagnetic bead-controlled test tube, gene detection reagent packaging, and gene detection method. Background Technology

[0002] Point-of-Care Testing (POCT) requires integrating multiple steps such as sample processing, nucleic acid extraction, washing, and amplification detection into a single closed device, achieving fully automated operation from sample input to result output. Nucleic acid extraction technology based on magnetic nanobeads is widely used in such integrated detection devices due to its ease of manipulation and elimination of the need for centrifugation. To ensure the sequential execution of different reaction steps, multiple functional chambers (such as sample processing chambers, washing chambers, and amplification chambers) are typically set up within the same test tube or chip. Before detection begins, reagents within each chamber must be kept isolated from each other, while an external magnetic field guides the magnetic nanobeads sequentially through each chamber to complete the transfer and detection of nucleic acids.

[0003] To address the aforementioned needs, existing technical solutions attempt to achieve isolation between the chambers by placing hydrophobic materials. For example, the inventor's authorized patent CN111876468A discloses a fully automated nucleic acid detection method and test tube, in which lysis buffer, washing buffer, and reaction solution are placed in horizontally arranged chambers, with the chambers isolated by solid or liquid hydrophobic materials. During detection, an external magnet controls nano-magnetic beads to sequentially pass through each hydrophobic isolation layer and finally enter the reaction chamber for amplification and detection. However, this technology still has the following significant shortcomings in practical applications and production processes:

[0004] (1) Problems when using solid hydrophobic materials (such as paraffin): Solid hydrophobic materials require the melting of thermoplastic materials such as paraffin wax during production, followed by injection into specific locations within a tube, and then solidification upon cooling to form an isolation layer. This process is complex and costly, and the solidified paraffin wax often necessitates the filling of gaps with a high-viscosity liquid, making the operation difficult. During testing, heating is required to melt the paraffin wax to allow magnetic beads to pass through; however, the melting of paraffin wax can easily lead to cross-contamination of liquids (such as pyrolysis fluid and cleaning fluid) between adjacent chambers, severely affecting the accuracy and repeatability of the test results.

[0005] (2) Problems when using liquid hydrophobic materials (such as oil phase): While liquid hydrophobic materials avoid the heating and melting process, they cannot be reliably fixed within the test tube. When the test tube is tilted or vibrated during transportation, storage, or use, the liquid hydrophobic material is prone to flow or displacement, leading to the failure of isolation between functional chambers, cross-contamination of liquids, and a high failure rate in testing.

[0006] (3) Limitations of the existing structure on the magnetic bead transfer path: In the above-mentioned horizontally arranged chamber structure, magnetic beads need to be continuously guided by an external magnetic field to pass through multiple isolation layers. However, the interfacial tension of hydrophobic materials may hinder the smooth passage of magnetic beads, especially when the solid hydrophobic material is not completely melted or the interface of the liquid hydrophobic material is unstable. Magnetic beads are prone to getting stuck or deviating from the path, reducing the nucleic acid recovery rate and detection sensitivity.

[0007] Therefore, there is an urgent need to develop a test tube device and corresponding gene detection method that can achieve stable, reliable, heat-free, and vibration-resistant isolation structure between functional chambers, while ensuring that the nanomagnetic beads can pass through each chamber sequentially with low resistance and without contamination, and is easy to manufacture and mass-produce. Summary of the Invention

[0008] In view of this, the present invention provides a method for packaging and detecting gene detection reagents using nanomagnetic beads in test tubes. By setting up isolation chambers and capillary channels, and forming a stable liquid isolation layer in the capillary channels with a high-viscosity isolation liquid, stable isolation between each chamber is achieved. At the same time, the nanomagnetic beads can be smoothly transferred between each chamber, thereby realizing the fully automated extraction, amplification and detection of nucleic acids.

[0009] To achieve the above objectives, the present invention adopts the following technical solution: A nano-magnetic bead controlled test tube, comprising: The test tube body includes a sample processing chamber, a washing chamber, and an amplification chamber. A sample isolation chamber and a capillary channel are provided between the sample processing chamber and the washing chamber. The sample isolation chamber has an opening communicating with the capillary channel. An amplification isolation chamber and a capillary channel are provided between the washing chamber and the amplification chamber. The amplification isolation chamber has an opening communicating with the capillary channel. High-viscosity isolation solutions are provided in both the sample isolation chamber and the amplification isolation chamber. The isolation solutions form a liquid isolation layer within the capillary channel. Solid or liquid amplification reagents are provided in the amplification chamber. Solid or liquid sample processing reagents and magnetic nanobeads are provided in the sample processing chamber.

[0010] Through the above technical solution, the present invention sets up a sample isolation chamber and capillary channel between the sample processing chamber and the cleaning chamber, and an amplification isolation chamber and capillary channel between the cleaning chamber and the amplification chamber. High-viscosity isolation liquid is injected into the sample isolation chamber and the amplification isolation chamber, forming a stable liquid isolation layer within the capillary channel. This achieves reliable isolation between the functional chambers, effectively avoiding the problem of liquid cross-flow between chambers during transportation, vibration, or testing. Simultaneously, this liquid isolation layer allows nano-magnetic beads to pass smoothly under the guidance of an external magnetic field without the need for heating and melting, overcoming the shortcomings of existing technologies where solid hydrophobic materials require heating and melting and are prone to cross-flow, and liquid hydrophobic materials are prone to flow.

[0011] Preferably, in the above-mentioned nanomagnetic bead controlled test tube, the washing chamber, the amplification chamber, the sample isolation chamber, the amplification isolation chamber, and the capillary channel are disposed at the bottom of the sample processing chamber; a magnetic bead enrichment chamber is disposed at the bottom of the sample processing chamber.

[0012] Preferably, in the above-mentioned nano-magnetic bead control test tube, the sample processing chamber has a sample inlet and a sample inlet sealing cap; the sample isolation chamber and the amplification isolation chamber are respectively provided with isolation injection holes; the cleaning chamber is provided with a cleaning injection hole and an exhaust hole; the amplification chamber is light-transmitting and is provided with a sealing hole.

[0013] Preferably, in the above-mentioned nanomagnetic bead controlled test tube, an isolation plate is provided between the sample processing chamber and the sample isolation chamber, the washing chamber, the amplification chamber, and the amplification isolation chamber; the isolation plate is provided with multiple sealing columns.

[0014] This invention also provides a method for packaging gene detection reagents, comprising: Amplification chamber encapsulation: Primer probe solution and reaction system are set at the bottom of the amplification chamber, and the primer probe solution and reaction system are sealed and isolated by solid or liquid hydrophobic material; the surface of the solid or liquid hydrophobic material is filled with high viscosity liquid material; Encapsulation of the cleaning chamber: High-viscosity isolation fluid is injected into the sample isolation chamber and amplification isolation chamber through the isolation injection port, forming a liquid isolation layer in the capillary channel; cleaning fluid of nano-magnetic beads is injected through the cleaning injection port; Sample processing chamber encapsulation: The sample processing system and nano magnetic beads are added inside the sample processing chamber and sealed with a sample dispensing port sealing cap.

[0015] Through the above technical solution, this invention uses a solid or liquid hydrophobic substance to seal the primer probe liquid and reaction system in the amplification chamber encapsulation step and fills its surface with a high-viscosity liquid substance. In the cleaning chamber encapsulation step, a high-viscosity isolation liquid is injected into the sample isolation chamber and the amplification isolation chamber to form a liquid isolation layer in the capillary channel. Then, the sealing posts on the isolation plate seal each injection hole and encapsulation hole, realizing the independent and stable encapsulation of reagents in each chamber. This method effectively prevents cross-contamination between reagents and displacement of hydrophobic substances during transportation and storage, ensures mutual isolation of liquids in each chamber before detection is started, and provides a stable liquid isolation interface for the smooth transfer of subsequent nanomagnetic beads.

[0016] This invention also provides a gene detection method, characterized by: adding a sample into a sample processing chamber, decomposing the sample to release nucleic acids through a sample processing system, and adsorbing the nucleic acids using magnetic nanobeads; setting a magnetic field outside the magnetic bead enrichment chamber, where the magnetic nanobeads are enriched under the influence of the magnetic field; transferring the magnetic field through an external device, where the magnetic nanobeads pass through the capillary channel corresponding to the sample isolation chamber and enter the washing chamber under the influence of the magnetic field; moving within the washing chamber and removing sample impurities under the action of the washing solution; the magnetic nanobeads continuing to pass through the capillary channel corresponding to the amplification isolation chamber and entering the amplification chamber, where the external magnetic field is removed; heating the amplification chamber using an external temperature control device, where the primer probe solution, reaction system, and magnetic nanobeads are thoroughly mixed to carry out the amplification reaction; and performing fluorescence detection of the amplification chamber using an external optical device.

[0017] Through the above technical solution, this invention directly adds the sample into the sample processing chamber, utilizes nano-magnetic beads to adsorb the released nucleic acid, and under the control of an external magnetic field, the magnetic beads automatically pass through the capillary channel of the sample isolation chamber into the washing chamber for washing, then through the capillary channel of the amplification isolation chamber into the amplification chamber, and finally, the amplification reaction is carried out by temperature-controlled heating and real-time fluorescence detection. This achieves a fully enclosed, fully automated integrated detection process from sample entry to result exit. This method eliminates the need for manual transfer of liquids or magnetic beads, avoiding sample cross-contamination and operational errors. At the same time, due to the presence of a liquid isolation layer in the capillary channel, the liquids in each chamber remain isolated during the transfer of magnetic beads, ensuring the accuracy and repeatability of the detection results. It is particularly suitable for molecular point-of-care detection scenarios.

[0018] Preferably, in the above-mentioned gene detection method, a magnetic field is provided at the bottom of the amplification cavity, and magnetic beads settle and adsorb at the bottom of the amplification cavity under the action of the magnetic field and gravity, and an optical device detects the side of the amplification cavity.

[0019] Preferably, in the above-mentioned gene detection method, the amplification reaction is a PCR amplification reaction or an isothermal amplification reaction.

[0020] As can be seen from the above technical solution, compared with the prior art, the present invention discloses a method for packaging and gene detection reagents using nanomagnetic beads in test tubes, which has the following beneficial effects: 1. Stable and reliable liquid isolation: By setting a high-viscosity isolation liquid in the sample isolation chamber and the amplification isolation chamber, and forming a stable liquid isolation layer in the capillary channel, effective isolation between the chambers is achieved, avoiding liquid cross-flow during transportation, vibration and detection. This overcomes the problems of solid hydrophobic materials needing to be heated and melted and being prone to cross-flow, and liquid hydrophobic materials being prone to flow in the prior art.

[0021] 2. Smooth transfer of magnetic beads and fully automated operation: The nano-magnetic beads can pass through the liquid isolation layer in the capillary channel in sequence under the guidance of an external magnetic field, without heating or additional operation. The transfer process is smooth and has low resistance. Combined with temperature-controlled amplification and fluorescence detection, it realizes a fully enclosed, fully automated integrated gene detection from sample in to result out.

[0022] 3. Compact structure and easy to manufacture: The magnetic bead enrichment chamber, cleaning chamber, amplification chamber, etc. are arranged sequentially in the circumference of the test tube body. Combined with the multi-hole sealing design of the sealing layer plate and the isolation plate, the structure is compact and highly integrated, which reduces the production and manufacturing costs and operational complexity.

[0023] 4. High accuracy and repeatability: The liquid isolation layer remains intact after the magnetic beads pass through, effectively preventing cross-contamination of reagents between chambers; a magnetic field can be set at the bottom of the amplification chamber to assist the sedimentation of magnetic beads, and side optical detection avoids interference from magnetic beads, ensuring the accuracy of fluorescence detection and the repeatability of results.

[0024] 5. Stable reagent packaging and strong applicability: The packaging method adopts a multi-layer seal of hydrophobic material and high viscosity liquid material to prevent reagents from shifting or becoming contaminated during transportation and storage; it supports two packaging forms: conventional liquid amplification reagents and lyophilized amplification reagents. The lyophilized solution can be transported at room temperature, reducing cold chain costs. Attached Figure Description

[0025] 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 embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0026] Figure 1 The attached figure is a schematic diagram of a half-section of the nano-magnetic bead controlled test tube provided by the present invention. Figure 2 The attached figure is a schematic diagram of the structure of the functional chamber layer of Embodiment 1 provided by the present invention; Figure 3The attached figure is a front view of the external structure of the test tube body provided by the present invention; Figure 4 The attached figure is provided by the present invention. Figure 3 Sectional view of AA.

[0027] in: 1-Sample processing chamber, 2-Magnetic bead enrichment chamber, 3-Capillary channel, 4-Sample isolation chamber, 5-Washing chamber, 6-Amplification isolation chamber, 7-Amplification chamber, 8-Test tube body; 9-Sealing plate, 10-Isolation plate; 11-Sample dispensing port, 12-Sample dispensing port sealing cap, 13-Washing injection hole, 14-Vent hole, 15-Sealing column, 16-Encapsulation hole; 17-Isolation injection hole; 18-Amplification chamber shell. Detailed Implementation

[0028] 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.

[0029] Example 1: See appendix Figure 1 To be continued Figure 4 This invention discloses a nano-magnetic bead controlled test tube, comprising: The test tube body 8 has a functional chamber layer at the bottom of its inner cavity and a sample processing chamber 1 above it. The sample processing chamber 1 is used to hold solid or liquid sample processing reagents and nano magnetic beads. The functional chamber layer includes a magnetic bead enrichment chamber 2, a sample isolation chamber 4, a washing chamber 5, an amplification isolation chamber 6, and an amplification chamber 7 arranged in a circumferential order. The magnetic bead enrichment chamber 2 is connected to the sample processing chamber 1 and is used to enrich the nano-magnetic beads with adsorbed nucleic acids under the action of an external magnetic field. Both the sample isolation chamber 4 and the amplification isolation chamber 6 form a capillary channel 3 with the inner wall of the test tube body 8, and both the sample isolation chamber 4 and the amplification isolation chamber 6 are provided with openings that communicate with the corresponding capillary channels 3; both the sample isolation chamber 4 and the amplification isolation chamber 6 are used to inject high-viscosity isolation liquid, and the isolation liquid forms a liquid isolation layer in the capillary channel 3; The cleaning chamber 5 is used to contain cleaning fluid to wash the nanomagnetic beads after they have passed through the sample isolation chamber 4, thereby removing sample impurities. Amplification chamber 7 is used to contain amplification reagents and to allow washed magnetic nanobeads to enter for nucleic acid amplification reaction and optical detection.

[0030] To further optimize the above technical solution, a sealing plate 9 is fastened to the upper part of the functional chamber layer. The sealing plate 9 has the following openings: a plate clearance opening for connecting the magnetic bead enrichment chamber 2 and the sample processing chamber 1, two isolation injection holes 17 corresponding to the sample isolation chamber 4 and the amplification isolation chamber 6 respectively, a cleaning injection hole 13 and an exhaust hole 14 corresponding to the cleaning chamber 5, and a packaging hole 16 corresponding to the amplification chamber 7.

[0031] To further optimize the above technical solution, an isolation plate 10 is fastened to the sealing plate 9. The isolation plate 10 has a sealing post 15 for sealing the isolation injection hole 17, the cleaning injection hole 13, the vent hole 14 and the sealing hole 16. The isolation plate 10 is provided with an isolation plate clearance opening corresponding to the plate clearance opening.

[0032] To further optimize the above technical solution, the bottom of the test tube body 8 has an amplification chamber shell 18 that extends downward and communicates with the amplification chamber 7.

[0033] To further optimize the above technical solution, the amplification chamber housing 18 is made of a light-transmitting material.

[0034] To further optimize the above technical solution, the top of the test tube body 8 is a sample inlet 11, and a sample inlet sealing cap 12 is screwed tightly onto the sample inlet 11.

[0035] The method for gene detection using magnetic bead-controlled test tubes provided in this embodiment includes the following steps: The sample is added into sample processing chamber 1, and the sample processing reagents decompose the sample to release nucleic acid, which is then adsorbed by nano magnetic beads. A magnetic field is applied outside the magnetic bead enrichment cavity 2, so that the nano-magnetic beads adsorbed with nucleic acids are enriched in the magnetic bead enrichment cavity 2 under the action of the magnetic field. The magnetic field is moved by an external device, so that the nano magnetic beads pass through the capillary channel 3 corresponding to the sample isolation cavity 4 and enter the cleaning cavity 5 under the guidance of the magnetic field. The nano-magnetic beads move within the cleaning chamber 5 and remove sample impurities under the action of the cleaning solution; Continue moving the magnetic field to allow the nanomagnetic beads to pass through the capillary channel 3 corresponding to the amplification isolation chamber 6 and enter the amplification chamber 7, and then remove the external magnetic field; The amplification chamber 7 is heated by an external temperature control device to ensure that the amplification reagents and nano magnetic beads are thoroughly mixed and the nucleic acid amplification reaction is carried out. Fluorescence detection of amplification chamber 7 is performed using an external optical device.

[0036] To further optimize the above technical solution, a magnetic field is set at the bottom of the amplification cavity 7, so that the nanomagnetic beads settle and adsorb at the bottom of the amplification cavity 7 under the dual action of the magnetic field and gravity, and the optical device performs fluorescence detection from the side of the amplification cavity 7.

[0037] To further optimize the above technical solution, the nucleic acid amplification reaction is either a PCR amplification reaction or an isothermal amplification reaction.

[0038] To further optimize the above technical solution, when the nanomagnetic beads pass through the capillary channel 3 corresponding to the sample isolation chamber 4 and the amplification isolation chamber 6, the liquid isolation layer formed by the high viscosity isolation liquid in the capillary channel 3 remains intact to prevent liquid cross-flow between the chambers.

[0039] Example 2: This embodiment provides a specific method for reagent encapsulation and gene detection using test tubes controlled by the aforementioned magnetic beads: I. Reagent Packaging Method (1) Encapsulation of amplification chamber: Primer probe liquid and reaction system are added to the bottom of amplification chamber 7 through encapsulation hole 16. Primer probe liquid and reaction system are sealed and isolated by solid or liquid hydrophobic material. High viscosity liquid material is then filled on the surface of solid or liquid hydrophobic material to prevent the hydrophobic material from shifting during transportation.

[0040] (2) Encapsulation of isolation chamber and cleaning chamber: High viscosity isolation liquid is injected into sample isolation chamber 4 and amplification isolation chamber 6 through isolation injection hole 17, so that the isolation liquid naturally forms a stable liquid isolation layer in capillary channel 3; nano magnetic bead cleaning liquid is injected into cleaning chamber 5 through cleaning injection hole 13, and exhaust hole 14 is used to exhaust gas in the chamber.

[0041] (3) Sealing: The isolation plate 10 is fastened to the sealing plate 9, so that the sealing column 15 blocks the isolation injection hole 17, the cleaning injection hole 13, the vent hole 14 and the sealing hole 16 respectively; at the same time, the isolation plate clearance port and the plate clearance port are kept connected to ensure that the magnetic bead enrichment cavity 2 and the sample processing cavity 1 are unobstructed.

[0042] (4) Sample processing chamber encapsulation: Add sample processing system and nano magnetic beads into sample processing chamber 1, and then tighten the sample dispensing port sealing cap 12 at the sample dispensing port 11 to complete the overall encapsulation.

[0043] II. Gene Detection Methods Add the sample to be tested (such as blood, saliva, swab washing solution, etc.) into the sample processing chamber 1 through the sample application port 11, and tighten the sample application port sealing cap 12. Gently invert to mix, the sample processing system (such as lysis buffer) decomposes the sample and releases nucleic acids, while the nano-magnetic beads adsorb the released nucleic acids.

[0044] The test tube is placed in an external magnetic field device, and a magnetic field is applied outside the magnetic bead enrichment cavity 2, so that the nano magnetic beads adsorbed with nucleic acid are enriched at the bottom of the magnetic bead enrichment cavity 2 under the action of the magnetic field.

[0045] An external device (such as a movable magnet or electromagnet) moves the magnetic field along the side wall of the test tube, guiding the nano-magnetic beads through the capillary channel 3 corresponding to the sample isolation chamber 4. Since the capillary channel 3 is pre-filled with a liquid isolation layer formed by a high-viscosity isolation liquid, the magnetic beads can pass through smoothly. At the same time, the liquid isolation layer quickly recovers its integrity after the magnetic beads pass through, preventing the liquid in the sample processing chamber 1 from entering the cleaning chamber 5.

[0046] After entering the washing chamber 5, the magnetic nanobeads move back and forth in the washing solution under the guidance of a magnetic field, thoroughly washing away sample impurities. The magnetic field continues to move, causing the magnetic nanobeads to pass through the capillary channel 3 corresponding to the amplification isolation chamber 6 and enter the amplification chamber 7, at which point the external magnetic field is removed.

[0047] The test tube is placed in an external temperature control device (such as a PCR instrument or isothermal amplification instrument) to heat amplification chamber 7, ensuring thorough mixing of the primer probe solution, reaction system, and magnetic nanobeads, and then a nucleic acid amplification reaction (such as PCR amplification or isothermal amplification) is performed. Simultaneously, real-time fluorescence detection of amplification chamber 7 is performed using an external optical device (such as a fluorescence detector), and the detection results are read.

[0048] Optionally, a magnetic field is set at the bottom of the amplification cavity 7, so that the nanomagnetic beads settle and adsorb at the bottom of the amplification cavity 7 under the combined action of the magnetic field and gravity. At this time, the optical device performs fluorescence detection from the side of the amplification cavity 7, which can effectively avoid the interference of the magnetic beads on the optical path.

[0049] Example 3: This embodiment is basically the same as Embodiment 2, except that the form of the reagent encapsulated in the amplification chamber 7 is different. In this embodiment, the bottom of the amplification chamber 7 is equipped with a lyophilized amplification reagent and a reconstitution solution containing specific primers, probes, enzymes, and dNTPs; the lyophilized amplification reagent and the reconstitution solution are sealed and isolated by a solid or liquid hydrophobic substance.

[0050] The specific packaging steps are as follows: (1) Encapsulation of amplification chamber: First, put the lyophilized amplification reagent ball or lyophilized powder into the bottom of amplification chamber 7, and then add the reconstitution solution; cover the lyophilized amplification reagent and the reconstitution solution with a layer of solid or liquid hydrophobic material (such as paraffin oil or low melting point wax) to isolate the amplification reagent from the outside world; fill the surface of the hydrophobic material with a layer of high viscosity liquid material to further prevent the hydrophobic material from flowing during transportation.

[0051] (2) The encapsulation of the remaining chambers is the same as in Example 1: high viscosity isolation liquid is injected into the sample isolation chamber 4 and amplification isolation chamber 6 through the isolation injection hole 17, cleaning liquid is injected through the cleaning injection hole 13, each hole is sealed by the sealing post 15 on the isolation plate 10, and finally the sample processing system and nano magnetic beads are added into the sample processing chamber 1 and sealed.

[0052] During testing, after the sample is added to sample processing chamber 1, the magnetic nanobeads sequentially pass through sample isolation chamber 4, washing chamber 5, and amplification isolation chamber 6 into amplification chamber 7. After the external magnetic field is removed, amplification chamber 7 is heated by an external temperature control device. During heating, the hydrophobic material melts or softens, and the lyophilized amplification reagent and reconstitution solution fully dissolve under the influence of temperature, mixing with the nucleic acids adsorbed on the magnetic nanobeads, triggering the amplification reaction. Simultaneously, an external optical device performs fluorescence detection on amplification chamber 7.

[0053] In this embodiment, the lyophilized amplification reagent has better room temperature stability, making it suitable for long-term storage and transportation without the need for a cold chain, further reducing detection costs and lowering the barrier to entry.

[0054] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on its differences from other embodiments. Similar or identical parts between embodiments can be referred to interchangeably. For the apparatus disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple; relevant parts can be referred to the method section.

[0055] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A test tube controlled by nanomagnetic beads, characterized in that, include: The test tube body (8) is provided with a sample processing chamber (1), a washing chamber (5) and an amplification chamber (7); a sample isolation chamber (4) and a capillary channel (3) are provided between the sample processing chamber (1) and the washing chamber (5); the sample isolation chamber (4) is provided with an opening that communicates with the capillary channel (3); an amplification isolation chamber (6) and a capillary channel (3) are provided between the washing chamber (5) and the amplification chamber (7); the amplification isolation chamber (6) is provided with an opening that communicates with the capillary channel (3); a high-viscosity isolation liquid is provided in the sample isolation chamber (4) and the amplification isolation chamber (6); the isolation liquid forms a liquid isolation layer in the capillary channel (3); a solid or liquid amplification reagent is provided in the amplification chamber (7); a solid or liquid sample processing reagent and nanomagnetic beads are provided in the sample processing chamber (1).

2. The nano-magnetic bead controlled test tube according to claim 1, characterized in that, The cleaning chamber (5), the amplification chamber (7), the sample isolation chamber (4), the amplification isolation chamber (6), and the capillary channel (3) are located at the bottom of the sample processing chamber (1); a magnetic bead enrichment chamber (2) is located at the bottom of the sample processing chamber (1).

3. The nano-magnetic bead controlled test tube according to claim 2, characterized in that, The sample processing chamber (1) has a sample inlet (11) and a sample inlet sealing cap (12); the sample isolation chamber (4) and the amplification isolation chamber (6) are respectively provided with isolation injection holes (17); the cleaning chamber (5) is provided with a cleaning injection hole (13) and an exhaust hole (14); the amplification chamber (7) is light-transmitting and is provided with a sealing hole (16).

4. The nano-magnetic bead controlled test tube according to claim 1, characterized in that, An isolation plate (10) is provided between the sample processing chamber (1), the sample isolation chamber (4), the cleaning chamber (5), the amplification chamber (7), and the amplification isolation chamber (6); the isolation plate (10) is provided with a plurality of sealing columns (15).

5. A method for packaging gene detection reagents, characterized in that, include: Encapsulation of amplification chamber (7): primer probe liquid and reaction system are provided at the bottom of amplification chamber (7), and the primer probe liquid and reaction system are sealed and isolated by solid or liquid hydrophobic material; the surface of the solid or liquid hydrophobic material is filled with high viscosity liquid material; Cleaning chamber (5) encapsulation: High viscosity isolation liquid is injected into the sample isolation chamber (4) and amplification isolation chamber (6) through the isolation injection hole (17) and a liquid isolation layer is formed in the capillary channel (3); Inject the cleaning solution of the nano-magnetic beads through the cleaning injection hole (13); Sample processing chamber (1) encapsulation: The sample processing system and nano magnetic beads are added to the sample processing chamber (1) and sealed by the sample filling port sealing cap (12).

6. A gene detection method, characterized in that: The sample is added into the sample processing chamber (1), and the sample is decomposed and nucleic acid is released through the sample processing system. The nucleic acid is adsorbed by the magnetic nanobeads. A magnetic field is set outside the magnetic bead enrichment chamber (2), and the magnetic nanobeads are enriched under the action of the magnetic field. The magnetic field is transferred by an external device. Under the action of the magnetic field, the nanomagnetic beads pass through the capillary channel (3) corresponding to the sample isolation chamber (4) and enter the cleaning chamber (5). The nanomagnetic beads move in the cleaning chamber (5) and remove sample impurities under the action of the cleaning solution. The nanomagnetic beads continue to pass through the capillary channel (3) corresponding to the amplification isolation chamber (6) and enter the amplification chamber (7) and then the external magnetic field is removed. The amplification chamber (7) is heated by an external temperature control device, and the primer probe solution, reaction system and nanomagnetic beads are thoroughly mixed to carry out the amplification reaction. The amplification chamber (7) is fluorescently detected by an external optical device.

7. A gene detection method according to claim 6, characterized in that, A magnetic field is provided at the bottom of the amplification cavity (7). The magnetic beads settle and are adsorbed at the bottom of the amplification cavity (7) under the action of the magnetic field and gravity. The optical device detects the side of the amplification cavity (7).

8. A gene detection method according to claim 6, characterized in that, The amplification reaction is either a PCR amplification reaction or an isothermal amplification reaction.

Images