Whole blood sample nucleic acid extraction card box

Abstract

The invention discloses a whole blood sample nucleic acid extraction card box. Comprising a clamping box base, and a splitting decomposition / combination chamber, a primary washing reagent pre-storage chamber, a secondary washing reagent pre-storage chamber, a drying chamber, an eluent pre-storage chamber, a magnetic bar sleeve pre-storage chamber, a slow release liquid pre-storage chamber, a pipette tip pre-storage chamber, a combination liquid pre-storage chamber and a sedimentation reagent pre-storage chamber which are respectively and movably arranged on the clamping box base, a whole blood collection unit and a pump valve assembly. Through a micro-liquid packaging technology, a whole blood sedimentation technology and a chamber detachable technology, micro-liquid is sealed in a hose, and sedimentation and transfer of whole blood and rapid assembly and disassembly of the whole card box are achieved while the micro-liquid is flushed into a target chamber by liquid with a large volume.

Description

Technical Field

[0001] The present invention relates to a technology in the field of medical equipment, in particular to a whole blood sample nucleic acid extraction card box with pump valve drive and pre-embedded reagents. Background Art

[0002] Existing nucleic acid detection technology is difficult to achieve on-site detection, and sample pretreatment has become its main limiting factor. Although the current related products have achieved the ultimate goal of sample in and result out, and have the prospect of on-site application, the product throughput is low, the cost is high, and it cannot directly process whole blood samples, which limits its application scenarios. Summary of the invention

[0003] In view of the above-mentioned deficiencies in the prior art, the present invention proposes a whole blood sample nucleic acid extraction card box, which has multiple pre-buried reagent chambers, which are connected by hoses, micro diaphragm pumps and two-way solenoid valves, and the directional flow of liquid is achieved by starting and stopping the pump and the valve.

[0004] The present invention is achieved through the following technical solutions:

[0005] The present invention relates to a nucleic acid extraction cartridge, comprising: a cartridge base and a lysis / binding chamber, a primary washing reagent pre-storage chamber, a secondary washing reagent pre-storage chamber, a drying chamber, an eluent pre-storage chamber, a magnetic rod sleeve pre-storage chamber, a slow-release liquid pre-storage chamber, a pipette tip pre-storage chamber, a binding liquid pre-storage chamber, a sedimentation reagent pre-storage chamber, a whole blood collection unit and a pump valve assembly.

[0006] The movable arrangement is designed to be detachable by providing a groove on the base of the card box.

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

[0008] The pump-valve assembly comprises: two micro-diaphragm pumps and four solenoid valves, wherein: the first micro-diaphragm pump is arranged between the lysis / combination chamber and the pre-storage chamber of the binding liquid, and the third solenoid valve, the second micro-diaphragm pump and the second and fourth solenoid valves in parallel are arranged in sequence between the lysis / combination chamber and the pre-storage chamber of the sedimentation reagent.

[0009] A lysis solution is pre-buried in the connecting pipe between the third solenoid valve and the second micro-diaphragm pump.

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

[0011] A magnetic bead liquid is pre-buried in the connecting tube between the lysis / binding chamber and the first micro-diaphragm pump. Technical Effects

[0012] The present invention uses trace liquid packaging technology, whole blood sedimentation technology and chamber detachable technology to achieve the encapsulation of trace liquid in a hose, which is flushed into the target chamber by a larger volume of liquid, while achieving the sedimentation and transfer of whole blood and the rapid loading and unloading of the entire card box. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is a schematic diagram of the present invention after the pump and valve are installed;

[0014] Figure 2 is a schematic diagram of the present invention;

[0015] Figure 3-4 It is a front view and a side view of the present invention;

[0016] Figure 5 It is a schematic diagram of a base of the present invention;

[0017] Figure 6 Cross-sectional view of the base for whole blood sample collection;

[0018] Figure 7 This is a schematic diagram of the whole blood sedimentation route;

[0019] In the figure: lysis / binding chamber 1, primary washing reagent pre-storage chamber 2, secondary washing reagent pre-storage chamber 3, drying chamber 4, eluent pre-storage chamber 5, magnetic rod sleeve pre-storage chamber 6, sustained-release liquid pre-storage chamber 7, pipette tip pre-storage chamber 8, binding liquid pre-storage chamber 9, whole blood collection tube support 10, collection needle 11, sedimentation reagent pre-storage chamber 12, cartridge base 13, whole blood collection base 14, first and second micro-diaphragm pumps 15, 16, first to fourth solenoid valves 17-20, lysis / binding chamber inlet 21, sedimentation reagent pre-storage chamber outlet 22 and inlet 23, sedimentation reagent pre-storage chamber sample inlet 24, whole blood sample outlet 25, whole blood sample bag 26;

[0020] Figure 8-Figure 10 A schematic diagram of an embodiment scenario. DETAILED DESCRIPTION

[0021] like Figure 1-Figure 3 As shown, a nucleic acid extraction cartridge involved in the present embodiment comprises: a cartridge base 13 and 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 eluent pre-storage chamber 5, a magnetic rod sleeve pre-storage chamber 6, a sustained-release liquid pre-storage chamber 7, a pipette tip pre-storage chamber 8, a binding liquid pre-storage chamber 9, a sedimentation reagent pre-storage chamber 12, a whole blood collection unit and a pump-valve assembly.

[0022] The lysis / combination chamber 1 is provided with a lysis / combination chamber liquid inlet 21 .

[0023] The sedimentation reagent pre-storage chamber 12 is provided with sedimentation reagent pre-storage chamber liquid inlet and outlet holes 22, 23 and a sedimentation reagent pre-storage chamber sample injection hole 24, wherein: the sedimentation reagent pre-storage chamber liquid outlet hole 22 and the liquid inlet hole 23 are respectively located at different depths in the sedimentation reagent pre-storage chamber 12, and the sedimentation reagent pre-storage chamber sample injection hole 24 is connected to the whole blood sample bag 26.

[0024] The whole blood collection unit comprises: a whole blood collection base 14 and a whole blood collection tube support 10 and a collection needle 11 arranged thereon.

[0025] The whole blood collection base 14 is provided with a whole blood sample outlet 25 .

[0026] like Figure 7 As shown, the pump-valve assembly includes: two micro-diaphragm pumps 15, 16 and four solenoid valves 17, 18, 19, 20, wherein: the first micro-diaphragm pump 15 is arranged between the lysis / combination chamber 1 and the binding liquid pre-storage chamber 9, the third solenoid valve 19, the second micro-diaphragm pump 16 and the second and fourth solenoid valves 18, 20 in parallel are arranged respectively between the lysis / combination chamber 1 and the sedimentation reagent pre-storage chamber 12.

[0027] This embodiment relates to a whole blood processing method of the above device, specifically comprising:

[0028] 1) Insert a blood collection tube (10 mL) into the whole blood collection tube holder 10;

[0029] 2) The control relay is started, and the second micro-diaphragm pump 16, the first solenoid valve 18, and the third solenoid valve 19 start to work. At this time, the blood in the whole blood sample bag 26 is pumped 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 with the whole blood entering the chamber, until it is higher than the lower end opening of the hose connected to the fourth solenoid valve 20. At the same time, the lysis liquid pre-buried in the hose between the third solenoid valve 19 and the second micro-diaphragm pump 16 is pushed by the gas to enter the lysis / combination chamber 1 in advance;

[0030] 3) After all the blood to be tested enters the sedimentation reagent pre-storage chamber 12, the relay is reset to a low level and left to stand for the sedimentation of the blood;

[0031] 4) Start the relay, the second micro-diaphragm pump 16, the third solenoid valve 19, and the fourth solenoid valve 20 start to work, at which time part of the reagent above the lower end opening of the liquid outlet hole 22 in the sedimentation reagent pre-storage chamber 12, i.e., plasma, is drawn out of the sedimentation reagent pre-storage chamber 12 by the action of the pump, and enters the lysis / binding chamber 1 together with the two liquids after passing through the connecting tube pre-buried with protease and lysate;

[0032] 5) After all the plasma is drawn out, the relay is reset. The heating film outside the lysis / binding chamber 1 is started to control the temperature and promote the lysis of the plasma;

[0033] 6) After the lysis is completed, the relay is started, and the first micro-diaphragm pump 15 and the first solenoid valve 17 start to work, and the reagents and the pre-buried magnetic bead liquid in the binding liquid chamber 9 are drawn into the lysis / binding chamber 1;

[0034] 7) After all the binding liquid has entered, the relay is reset. The magnetic bead liquid completes the binding with the sample in step 5 in the lysis / binding chamber 1;

[0035] 8) Transfer the magnetic beads into the primary washing reagent pre-storage chamber 2 by adsorption of the magnetic rod, about 2 mL, demagnetize and mix for 30 seconds, insert the magnetic rod to adsorb the magnetic beads for 1 minute, and fully adsorb the magnetic beads;

[0036] 9) Transfer the magnetic beads to the secondary washing reagent pre-storage chamber 3 by magnetic adsorption, about 1 mL, demagnetize and mix for 30 minutes. Insert the magnetic rod to adsorb the magnetic beads for 1 minute to fully adsorb the magnetic beads;

[0037] 10) The magnetic beads are transferred to the drying chamber 4 by adsorption by the magnetic rod and warm-dried at 37°C for about 5 minutes (the time can be adjusted and set, and the adjustment time process needs to be judged by the naked eye until there is no water reflection on the surface of the magnetic beads);

[0038] 11) The magnetic beads are transferred by the magnetic rod into the pre-storage chamber 5 of the eluent, about 200uL, mixed for 30 minutes after demagnetization, allowed to stand for 3 minutes, and the magnetic rod is inserted to adsorb the magnetic beads for 1 minute to fully adsorb the magnetic beads;

[0039] 12) Transferring the magnetic rod sleeve to the magnetic rod sleeve pre-storage chamber 6;

[0040] 13) The pipette tip is installed in the pipette tip pre-storage chamber 8;

[0041] 14) The pipette sequentially transfers the nucleic acid solution in the eluent pre-storage chamber 5 and the slow-release solution in the slow-release solution pre-storage chamber 7 for subsequent detection until a DNA sample is obtained.

[0042] like Figure 8-Figure 10 As shown, the present invention adopts a detachable design, which can realize the pre-embedding of liquid and the unified treatment after the reaction is completed, and can directly process whole blood samples.

[0043] The above-mentioned specific implementation can be partially adjusted in different ways by those skilled in the art without departing from the principle and purpose of the present invention. The protection scope of the present invention shall be based on the claims and shall not be limited by the above-mentioned specific implementation. Each implementation scheme within its scope shall be subject to the constraints of the present invention.

Claims

1. A nucleic acid extraction cartridge, characterized in that: include: A cartridge base and a lysis / binding chamber, a primary washing reagent pre-storage chamber, a secondary washing reagent pre-storage chamber, a drying chamber, an eluent pre-storage chamber, a magnetic rod sleeve pre-storage chamber, a sustained-release liquid pre-storage chamber, a pipette tip pre-storage chamber, a binding liquid pre-storage chamber, a sedimentation reagent pre-storage chamber, a whole blood collection unit and a pump-valve assembly, which are movably arranged on the cartridge base.

2. The nucleic acid extraction cartridge according to claim 1, characterized in that: The movable arrangement is designed to be detachable by providing a groove on the base of the card box.

3. The nucleic acid extraction cartridge according to claim 1, wherein: The sedimentation reagent pre-storage chamber is provided with a sedimentation reagent pre-storage chamber liquid inlet and outlet holes and a sedimentation reagent pre-storage chamber sample injection hole, wherein: the sedimentation reagent pre-storage chamber liquid outlet hole and liquid inlet hole are respectively located at different depths in the sedimentation reagent pre-storage chamber, and the sedimentation reagent pre-storage chamber sample injection hole is connected to the whole blood sample bag.

4. The nucleic acid extraction cartridge according to claim 1, wherein: The pump-valve assembly comprises: two micro-diaphragm pumps and four solenoid valves, wherein: the first micro-diaphragm pump is arranged between the lysis / combination chamber and the pre-storage chamber of the binding liquid, and the third solenoid valve, the second micro-diaphragm pump and the second and fourth solenoid valves in parallel are arranged in sequence between the lysis / combination chamber and the pre-storage chamber of the sedimentation reagent.

5. The nucleic acid extraction cartridge according to claim 4, characterized in that: A lysis solution is pre-buried in the connecting pipe between the third solenoid valve and the second micro-diaphragm pump; Proteinase K is pre-buried in the connecting pipe between the fourth solenoid valve and the second micro-diaphragm pump; A magnetic bead liquid is pre-buried in the connecting tube between the lysis / binding chamber and the first micro-diaphragm pump.

6. A whole blood processing method based on the device according to any one of claims 1 to 5, characterized in that: include: 1) Insert the blood collection tube into the whole blood collection tube holder; 2) The control relay is started, and the second micro-diaphragm pump, the first solenoid valve, and the third solenoid valve start working. At this time, the blood in the whole blood sample bag is pumped 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 with the whole blood until it is higher than the lower end opening of the hose connected to the fourth solenoid valve. At the same time, the lysis liquid pre-buried in the hose between the third solenoid valve and the second micro-diaphragm pump is pushed by the gas to enter the lysis / combination chamber in advance; 3) After all the blood to be tested enters the sedimentation reagent pre-storage chamber, the relay is reset to a low level and left to stand for the sedimentation of the blood; 4) Start the relay, the second micro-diaphragm pump, the third solenoid valve, and the fourth solenoid valve start to work, at which time part of the reagent above the lower end opening of the liquid outlet hole in the sedimentation reagent pre-storage chamber, i.e., plasma, is drawn out of the sedimentation reagent pre-storage chamber by the action of the pump, and enters the lysis / binding chamber together with the two liquids after passing through the connecting tube pre-buried with protease and lysis solution; 5) After all the plasma is drawn out, the relay is reset, and the heating membrane outside the lysis / binding chamber is started to control the temperature and promote the lysis of the plasma; 6) After the lysis is completed, the relay is started, and the first micro-diaphragm pump and the first solenoid valve start to work, and the reagents in the binding liquid chamber and the pre-buried magnetic bead liquid are extracted into the lysis / binding chamber; 7) After all the binding liquid has entered, the relay is reset, and the magnetic bead liquid completes the binding with the sample in step 5 in the lysis / binding chamber. 8) The magnetic beads are transferred by the magnetic rod into the pre-storage chamber of the primary washing reagent, mixed after demagnetization, and the magnetic rod is inserted to adsorb the magnetic beads; 9) The magnetic beads are transferred by the magnetic rod into the secondary washing reagent pre-storage chamber, mixed after demagnetization, and the magnetic rod is inserted to adsorb the magnetic beads; 10) The magnetic beads are transferred by the magnetic rod into the drying room for warm drying; 11) The magnetic beads are transferred into the pre-storage chamber of the eluent by adsorption by the magnetic rod, mixed and allowed to stand after demagnetization, and the magnetic rod is inserted to adsorb the magnetic beads; 12) Transferring 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 pipette sequentially transfers the nucleic acid solution in the elution solution pre-storage chamber and the slow-release solution in the slow-release solution pre-storage chamber for subsequent testing until a DNA sample is obtained.

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