Whole blood sample nucleic acid extraction cartridge

Abstract

A whole blood sample nucleic acid extraction cartridge includes: a cartridge base and, respectively movably disposed on the cartridge base, a lysis / binding chamber, a primary wash reagent pre-storage chamber, a secondary wash reagent pre-storage chamber, a drying chamber, an eluent pre-storage chamber, a magnetic rod sleeve pre-storage chamber, a sustained-release solution pre-storage chamber, a pipette tip pre-storage chamber, a binding solution pre-storage chamber, a sedimentation reagent pre-storage chamber, a whole blood collection unit, and a pump valve assembly. This invention utilizes micro-liquid encapsulation technology, whole blood sedimentation technology, and chamber detachability technology to achieve the simultaneous sedimentation and transfer of whole blood and rapid loading and unloading of the entire cartridge, while a small amount of liquid is sealed within a flexible tube and flushed into the target chamber by a larger volume of liquid.

Description

Technical Field

[0001] This invention relates to a technology in the field of medical devices, specifically a whole blood sample nucleic acid extraction cartridge with a pump valve drive and pre-embedded reagents. Background Technology

[0002] Current nucleic acid testing technologies are difficult to implement on-site, with sample pretreatment being a major limiting factor. While existing products achieve the ultimate goal of sample input and result output, showing promise for on-site applications, their throughput is low, costs are high, and they cannot directly process whole blood samples, thus limiting their application scenarios. Summary of the Invention

[0003] To address the aforementioned shortcomings of existing technologies, this invention proposes a whole blood sample nucleic acid extraction cartridge. This cartridge utilizes multiple pre-embedded reagent chambers connected by tubing, a micro diaphragm pump, and a two-way solenoid valve. The directional flow of liquid is achieved by starting and stopping the pump and valve.

[0004] This invention is achieved through the following technical solution:

[0005] This invention relates to a nucleic acid extraction cartridge, comprising: a cartridge base and movably disposed on the cartridge base, a lysis / binding chamber, a primary washing reagent pre-storage chamber, a secondary washing reagent pre-storage chamber, a drying chamber, an elution buffer pre-storage chamber, a magnetic rod sleeve pre-storage chamber, a sustained-release solution pre-storage chamber, a pipette tip pre-storage chamber, a binding solution pre-storage chamber, a sedimentation reagent pre-storage chamber, a whole blood collection unit, and a pump valve assembly.

[0006] The aforementioned activity setup features a detachable design achieved through a groove on the card holder base.

[0007] The whole blood collection unit includes: a whole blood collection base and a whole blood collection tube support and a collection needle mounted thereon.

[0008] The pump and valve assembly includes two micro diaphragm pumps and four solenoid valves, wherein the first micro diaphragm pump and the first solenoid valve are sequentially disposed between the lysis / binding chamber and the binding solution pre-storage chamber, and the third solenoid valve, the second micro diaphragm pump, and the second and fourth solenoid valves connected in parallel are sequentially disposed between the lysis / binding chamber and the sedimentation reagent pre-storage chamber.

[0009] The connecting pipe between the third solenoid valve and the second micro diaphragm pump is pre-embedded with pyrolysis fluid.

[0010] Proteinase K is pre-embedded in the connecting pipe between the fourth solenoid valve and the second micro diaphragm pump.

[0011] The connecting pipe between the pyrolysis / binding chamber and the first micro diaphragm pump is pre-embedded with magnetic beads.

[0012] Technical effect

[0013] This invention utilizes micro-liquid encapsulation technology, whole blood sedimentation technology, and detachable chamber technology to achieve the simultaneous encapsulation of micro-liquid in a tubing, allowing it to be flushed into the target chamber by a larger volume of liquid, while also enabling the sedimentation and transfer of whole blood and the rapid loading and unloading of the entire cartridge. Attached Figure Description

[0014] Figure 1 This is a schematic diagram of the present invention after the pump and valve are installed;

[0015] Figure 2 This is a schematic diagram of the present invention;

[0016] Figure 3-4 These are the front view and side view of the present invention;

[0017] Figure 5 This is a schematic diagram of the base of the present invention;

[0018] Figure 6 A cross-sectional view of the base for whole blood sample collection;

[0019] Figure 7 This is a schematic diagram of the whole blood sedimentation pathway;

[0020] In the diagram: 1. Lysis / binding chamber; 2. Pre-storage chamber for primary washing reagent; 3. Pre-storage chamber for secondary washing reagent; 4. Drying chamber; 5. Pre-storage chamber for elution buffer; 6. Pre-storage chamber for magnetic rod sleeve; 7. Pre-storage chamber for sustained-release solution; 8. Pre-storage chamber for pipette tip; 9. Pre-storage chamber for binding solution; 10. Whole blood collection tube support; 11. Collection needle; 12. Pre-storage chamber for sedimentation reagent; 13. Cartridge base; 14. Whole blood collection base; 15, 16. First and second micro diaphragm pumps; 17-20. First to fourth solenoid valves; 21. Inlet port for lysis / binding chamber; 22. Outlet port and inlet port for sedimentation reagent pre-storage chamber; 23. Sample inlet port for sedimentation reagent pre-storage chamber; 24. Whole blood sample outlet port; 25. Whole blood sample bag; 26.

[0021] Figures 8-10 This is a schematic diagram of an example scenario. Detailed Implementation

[0022] like Figures 1-3 As shown, this embodiment relates to a nucleic acid extraction cartridge, which includes: a cartridge base 13 and grooves respectively disposed on the cartridge base 13 for a lysis / binding chamber 1, a primary washing reagent pre-storage chamber 2, a secondary washing reagent pre-storage chamber 3, a drying chamber 4, an elution buffer pre-storage chamber 5, a magnetic rod sleeve pre-storage chamber 6, a sustained-release solution pre-storage chamber 7, a pipette tip pre-storage chamber 8, a binding solution pre-storage chamber 9, a sedimentation reagent pre-storage chamber 12, a whole blood collection unit, and a pump valve assembly.

[0023] The pyrolysis / binding chamber 1 is provided with a pyrolysis / binding chamber liquid inlet 21.

[0024] The sedimentation reagent pre-storage chamber 12 is provided with sedimentation reagent pre-storage chamber inlet and outlet holes 22 and 23 and sedimentation reagent pre-storage chamber sample inlet hole 24, wherein: sedimentation reagent pre-storage chamber outlet hole 22 and inlet hole 23 are located at different depths in sedimentation reagent pre-storage chamber 12, and sedimentation reagent pre-storage chamber sample inlet hole 24 is connected to whole blood sample bag 26.

[0025] The whole blood collection unit includes a whole blood collection base 14 and a whole blood collection tube support 10 and a collection needle 11 disposed thereon.

[0026] The whole blood collection base 14 is provided with a whole blood sample outlet 25.

[0027] like Figure 7 As shown, the pump and valve assembly includes: two micro diaphragm pumps 15 and 16 and four solenoid valves 17, 18, 19 and 20, wherein: the first micro diaphragm pump 15 and the first solenoid valve 17 are sequentially arranged between the lysis / binding chamber 1 and the binding liquid pre-storage chamber 9, and the third solenoid valve 19, the second micro diaphragm pump 16 and the second and fourth solenoid valves 18 and 20 connected in parallel are sequentially arranged between the lysis / binding chamber 1 and the sedimentation reagent pre-storage chamber 12.

[0028] This embodiment relates to a whole blood processing method using the aforementioned device, specifically including:

[0029] 1) Insert a blood collection tube (10 mL) into the whole blood collection tube stent 10;

[0030] 2) When the control relay is activated, the second micro diaphragm pump 16, the second solenoid valve 18, and the third solenoid valve 19 start working. At this time, the blood in the whole blood sample bag 26 is drawn into the sedimentation reagent pre-storage chamber 12 due to the action of the pump. The liquid originally contained in the sedimentation reagent pre-storage chamber 12 rises as the whole blood enters, until it is higher than the lower opening of the tubing connected to the fourth solenoid valve 20. At the same time, the lysis solution pre-embedded in the tubing between the third solenoid valve 19 and the second micro diaphragm pump 16 is pushed by the gas into the lysis / binding chamber 1.

[0031] 3) After all the blood to be tested has entered the sedimentation reagent pre-storage chamber 12, the relay is reset to low level and left to stand for blood sedimentation.

[0032] 4) Start the relay, and the second micro diaphragm pump 16, the third solenoid valve 19, and the fourth solenoid valve 20 will start working. At this time, the part of the reagent above the lower opening of the sedimentation reagent pre-storage chamber 22 in the sedimentation reagent pre-storage chamber 12, i.e. plasma, will be pumped out of the sedimentation reagent pre-storage chamber 12 by the action of the pump. After passing through the connecting tube pre-embedded with protease and lysis buffer, the two liquids will enter the lysis / binding chamber 1 together.

[0033] 5) After all the plasma has been extracted, the relay is reset. The heating membrane outside the lysis / binding chamber 1 is activated to control the temperature and promote plasma lysis;

[0034] 6) After pyrolysis is completed, the relay is activated, the first micro diaphragm pump 15 and the first solenoid valve 17 start working, and the reagent and the pre-embedded magnetic bead liquid in the pre-storage chamber 9 are extracted into the pyrolysis / binding chamber 1.

[0035] 7) After all the binding solution has entered, the relay is reset. The magnetic bead solution completes its binding with the sample from step 5 in the lysis / binding chamber 1;

[0036] 8) The magnetic beads are transferred into the pre-storage chamber of the washing reagent by the magnetic rod. After demagnetization, mix well for 30 seconds, insert the magnetic rod to adsorb the magnetic beads for 1 minute to fully adsorb the magnetic beads.

[0037] 9) The magnetic beads are transferred into the secondary washing reagent pre-storage chamber 3 by magnetic rod adsorption and about 1 mL. After demagnetization, mix well for 30 minutes, insert the magnetic rod to adsorb the magnetic beads for 1 minute to fully adsorb the magnetic beads.

[0038] 10) The magnetic beads are transferred by magnetic rod into drying chamber 4 and dried at 37°C for about 5 minutes (the time can be adjusted and set, and the adjustment process needs to be judged by the naked eye until there is no water reflection on the surface of the magnetic beads).

[0039] 11) The magnetic beads are transferred into the elution buffer pre-storage chamber 5 (approximately 200 μL) by magnetic rod adsorption. After demagnetization, mix well for 30 minutes, let stand for 3 minutes, insert the magnetic rod to adsorb the magnetic beads for 1 minute, and fully adsorb the magnetic beads.

[0040] 12) Transfer the magnetic rod sleeve to the magnetic rod sleeve pre-storage chamber 6;

[0041] 13) The pipette tip is installed in the pipette tip pre-storage chamber 8;

[0042] 14) The nucleic acid solution in the elution buffer pre-storage chamber 5 and the sustained release solution in the sustained release buffer pre-storage chamber 7 are transferred sequentially by pipette for subsequent testing until a DNA sample is obtained.

[0043] like Figures 8-10 As shown, the present invention adopts a detachable design, which can realize the pre-embedding of liquids and unified processing after the reaction, and can directly process whole blood samples.

[0044] The above-described specific implementations can be partially adjusted by those skilled in the art in different ways without departing from the principles and purpose of the present invention. The scope of protection of the present invention is defined by the claims and is not limited to the above-described specific implementations. All implementation schemes within the scope of the claims are bound by the present invention.

Claims

1. A nucleic acid extraction cartridge, characterized in that, include: The cartridge base and the following components are movably mounted on the cartridge base: lysis / binding chamber, primary wash reagent pre-storage chamber, secondary wash reagent pre-storage chamber, drying chamber, eluent pre-storage chamber, magnetic rod sleeve pre-storage chamber, sustained-release solution pre-storage chamber, pipette tip pre-storage chamber, binding solution pre-storage chamber, sedimentation reagent pre-storage chamber, whole blood collection unit, and pump valve assembly. The pump and valve assembly includes two micro diaphragm pumps and four solenoid valves, wherein: a first solenoid valve and a first micro diaphragm pump are arranged sequentially from the binding liquid pre-storage chamber to the lysis / binding chamber, and a third solenoid valve, a second micro diaphragm pump, and a second and fourth solenoid valve connected in parallel are arranged sequentially from the lysis / binding chamber to the sedimentation reagent pre-storage chamber. The connecting pipe between the third solenoid valve and the second micro diaphragm pump is pre-embedded with pyrolysis fluid. Proteinase K is pre-embedded in the connecting pipe between the fourth solenoid valve and the second micro diaphragm pump. The connecting pipe between the pyrolysis / binding chamber and the first micro diaphragm pump is pre-embedded with magnetic beads.

2. The nucleic acid extraction cartridge according to claim 1, characterized in that, The aforementioned activity setup features a detachable design achieved through a groove on the card holder base.

3. The nucleic acid extraction cartridge according to claim 1, characterized in that, The sedimentation reagent pre-storage chamber is provided with a sedimentation reagent pre-storage chamber inlet / outlet port and a sedimentation reagent pre-storage chamber inlet port, wherein: the sedimentation reagent pre-storage chamber outlet port and the sedimentation reagent pre-storage chamber inlet port are located at different depths within the sedimentation reagent pre-storage chamber, and the sedimentation reagent pre-storage chamber inlet port is connected to the whole blood sample bag.

4. The nucleic acid extraction cartridge according to claim 3, characterized in that, The whole blood collection unit includes: a whole blood collection base and a whole blood collection tube support and a collection needle mounted thereon.

5. A method for whole blood processing based on the nucleic acid extraction cartridge of claim 4, characterized in that, include: 1) Insert the blood collection tube into the whole blood collection tube stent; 2) When the control relay is activated, the second micro diaphragm pump, the second solenoid valve, and the third solenoid valve start working. At this time, the blood in the whole blood sample bag is drawn into the sedimentation reagent pre-storage chamber due to the action of the pump. The liquid originally contained in the sedimentation reagent pre-storage chamber rises as the whole blood enters, until it is higher than the lower opening of the tubing connected to the fourth solenoid valve. At the same time, the lysis solution pre-embedded in the tubing between the third solenoid valve and the second micro diaphragm pump is pushed into the lysis / binding chamber by the gas. 3) After all the blood to be tested has entered the sedimentation reagent pre-storage chamber, the relay is reset to low level and left to stand for blood sedimentation. 4) Start the relay, and the second micro diaphragm pump, the third solenoid valve, and the fourth solenoid valve will start working. At this time, the part of the reagent above the lower opening of the liquid outlet of the sedimentation reagent pre-storage chamber, i.e. plasma, will be pumped out of the sedimentation reagent pre-storage chamber by the action of the pump and enter the lysis / binding chamber together with the two liquids through the connecting tube pre-embedded with protease and lysis solution. 5) After all the plasma has been extracted, the relay is reset, and the heating membrane outside the lysis / binding chamber is activated to control the temperature and promote the lysis of the plasma. 6) After pyrolysis is completed, the relay is activated, the first micro diaphragm pump and the first solenoid valve start working, and the reagents and pre-embedded magnetic beads in the pre-stored binding solution chamber are drawn into the pyrolysis / binding chamber. 7) After all the binding solution has entered, the relay is reset, and the magnetic bead solution completes its binding with the sample from step 5 in the lysis / binding chamber. 8) The magnetic beads are transferred into the pre-storage chamber of the washing reagent by magnetic rod adsorption. After demagnetization, they are mixed and then the magnetic beads are adsorbed by inserting the magnetic rod. 9) The magnetic beads are transferred into the secondary washing reagent pre-storage chamber by magnetic rod adsorption. After demagnetization, they are mixed and then the magnetic rod is inserted to adsorb the magnetic beads. 10) Magnetic beads are transferred by magnetic rods into the drying chamber for warm drying; 11) The magnetic beads are transferred into the eluent pre-storage chamber by magnetic rod adsorption. After demagnetization, they are mixed and allowed to stand. Then, the magnetic rod is inserted to adsorb the magnetic beads. 12) Transfer the magnetic rod sleeve to the magnetic rod sleeve pre-storage chamber; 13) The pipette tip is installed in the pipette tip pre-storage chamber; 14) The nucleic acid solution in the elution buffer pre-storage chamber and the sustained release solution in the sustained release buffer pre-storage chamber are transferred sequentially by pipette for subsequent testing until a DNA sample is obtained.

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