Sample pretreatment equipment and its reagent cartridges
By designing a well-sealed test reagent cartridge and utilizing the rotational and lifting motion of the liquid storage turntable and pipetting device, a fully enclosed operation is achieved, solving the problems of contamination and high cost in molecular diagnostic equipment. This supports the simultaneous detection of multiple pathogens and improves the accuracy and operability of the test.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- GUANGZHOU WEIYUAN MEDICAL INSTR CO LTD
- Filing Date
- 2022-06-15
- Publication Date
- 2026-05-26
AI Technical Summary
In existing molecular diagnostic equipment, the pathogen detection process is easily contaminated, and disposable detection devices are costly, complex in construction, and difficult to detect multiple pathogens simultaneously.
Design a well-sealed test reagent cartridge, comprising a liquid storage disc, a pipetting device, and a cartridge body. It achieves fully enclosed operation through rotation and lifting movements, supports simultaneous detection of multiple reagents, avoids cross-contamination, and eliminates the need for pre-sealed reagents through the rotation of the liquid storage disc and the coordination of the sample inlet.
It achieves a fully enclosed sample pretreatment process, avoiding contamination, reducing operational difficulty and cost, supporting simultaneous detection of multiple samples, and improving detection accuracy and operability.
Smart Images
Figure CN114907962B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of molecular diagnostic equipment technology, and in particular to a sample pretreatment device and a reagent cartridge therein. Background Technology
[0002] The detection process of pathogens in molecular diagnostics requires the lysis, washing, elution, separation and purification of pathogen nucleic acids. The purification process is complicated, time-consuming and has low extraction efficiency. If the above steps are carried out in an open environment, it is easy to generate aerosols, splashes and other contamination between samples, or medical staff may be infected by pathogens. It will also affect the purity of the extract and thus affect the accuracy of the experimental results.
[0003] For the reasons mentioned above, there are also some studies on closed cartridges in conventional technologies. For example, in molecular diagnostic equipment technology, multi-step pathogen detection methods have been integrated into one system. Users only need to add the sample to the system, and the remaining steps are all completed automatically by the system.
[0004] Therefore, molecular diagnostic cartridges and microfluidic chips use sealed, single-use test cards or tubes, which effectively control cross-contamination and carryover. However, these test cards or tubes often can only detect one pathogen at a time, thus requiring a large number of instruments to be carried, which is inconvenient for on-site operations.
[0005] In conventional technologies, there are also some integrated pathogen detection kits that can extract, amplify, and detect the nucleic acid to be tested in a closed system, which greatly avoids the contamination of the sample during the extraction and amplification process and effectively improves the accuracy and operability of nucleic acid detection. However, these detection devices generally have problems such as complex structure, high manufacturing cost, and high detection cost. Summary of the Invention
[0006] Therefore, it is necessary to provide a test reagent cartridge to address the above problems. This cartridge has the advantages of good sealing, can detect multiple samples simultaneously, is compatible with multiple reagents, and is open and compatible with various reagents. It does not require the reagents to be sealed in the cartridge, is universal for different reagents, and will not cause cross-infection, thus solving the problems of pathogenic microorganism contamination and operator infection.
[0007] A test kit, comprising:
[0008] A liquid storage turntable is provided with a plurality of liquid tanks for containing liquid and sample tube holes for containing sample tubes, wherein the sample tube holes and the plurality of liquid tank samples are arranged around the rotation axis of the liquid storage turntable.
[0009] A pipetting apparatus, comprising a pipette for aspirating and dispensing liquid, wherein the position of the pipette tip corresponds to the position of the liquid tank; and
[0010] The container has a liquid storage turntable installed inside, dividing the internal space into an upper cavity above the turntable surface and a lower cavity below it. The turntable can rotate and move up and down along the inner wall of the container. The pipetting device is fixed to the container, with the pipetting head located in the upper cavity. The turntable and the inner wall of the container form a sealed space in the upper cavity. The container has a sample inlet communicating with the upper cavity and a matching cover for the sample inlet.
[0011] The aforementioned test reagent cartridge is equipped with a lid. After all sample is added through the sample inlet, the cartridge can be tightly closed. Subsequent sample pretreatment is completed inside the cartridge, ensuring a sealed and contamination-free process. Furthermore, the test reagent cartridge can be adjusted for different testing applications by rotating the liquid storage disc and engaging the sample inlet, eliminating the need for pre-sealed reagents inside the cartridge.
[0012] In one embodiment, the liquid tank includes a reagent tank, a reaction tank, and a waste liquid tank. It is understood that the number and function of each type of tank within the specific liquid tank can be configured according to the requirements of the specific testing project. For example, a larger waste liquid tank can be designed for testing projects that generate a large amount of waste liquid.
[0013] In one embodiment, several liquid tanks and sample tube openings are arranged in a ring around the axis of the liquid storage turntable, with the pipette tip and sample inlet corresponding to the positions of the ring. With this arrangement, the pipette and its pipette tip can be fixed in place, and the rotation of the liquid storage turntable ensures that all liquid tank positions and sample tube openings correspond to the fixed pipette tip, avoiding aspiration errors or operational difficulties caused by moving the pipette tip. In conventional technologies, the pipette tip is usually separate from the aspiration device and connected by a flexible tube, which carries the risk of tube breakage during pipette tip movement, leading to insufficient pipetting accuracy or incomplete aspiration.
[0014] In one embodiment, the pipette is a piston-type pipette. By fixing the pipette tip and integrating the pipette piston into the overall design, external actuation is simplified; only the piston needs to be moved in a single direction, offering advantages such as reliable and stable operation. It is understood that the piston-type pipette can be made from devices such as syringes, and the required accuracy can be adjusted by controlling the tube diameter, i.e., the volume of liquid that can be transferred by moving the piston a certain distance.
[0015] In one embodiment, two pipettes are provided, and the liquid storage disc is also provided with a clearance hole. When one pipette corresponds to the position of the liquid tank, the other pipette corresponds to the position of the clearance hole. It is understood that since the position of the pipettes remains unchanged when the liquid storage disc moves upward, if there are two pipettes, and one pipette is used while the other is idle, if the idle pipette has no corresponding hole, it will collide with the surface of the liquid storage disc. Therefore, clearance design for the pipettes needs to be considered. The specific location of the clearance hole can be determined by those skilled in the art based on the specific testing procedure requirements, as well as the number and location of the applicable pipettes. The clearance hole can be a specially designed hole or another reagent tank that does not affect the testing procedure.
[0016] It is also understood that the number of pipette tips is not limited. One pipette tip can complete all processes, two pipette tips can effectively prevent impurities from the sample from being carried into the pure sample, or more pipette tips can also complete all processes. However, the number of pipette tips will affect the size of the liquid storage turntable and the number of clearance holes, thus affecting the size of the entire cartridge. It can be set according to the specific testing requirements.
[0017] In one embodiment, the inner wall of the box is further provided with a limiting member for restricting the downward movement of the liquid storage turntable, and the limiting member is disposed in the lower cavity. The limiting member is used to prevent the liquid storage turntable from falling out of the box due to excessive downward movement. It is understood that the specific shape and position of the limiting member can be set according to conventional techniques in the art.
[0018] In one embodiment, a rotary sealing ring is provided between the liquid storage rotary plate and the inner wall of the box, forming a dynamic seal between the liquid storage rotary plate and the box. It is understood that the sealing requirements of the upper cavity can also be achieved by other means in the art, but achieving a dynamic seal through a rotary sealing ring has the advantages of being convenient and reliable, and not affecting the movement of the liquid storage rotary plate.
[0019] In one embodiment, the test kit further includes a guide bracket, through which the liquid storage disc is mounted within the kit body, and a sealing ring for the disc is fitted around the outer periphery of the guide bracket. It is understood that using a guide bracket to guide and move the liquid storage disc offers advantages such as stable movement and convenient installation.
[0020] In one embodiment, the test kit further includes a sealing cap for sealing the liquid storage disc. The sealing cap is mounted on the guide bracket, and the liquid storage disc can rotate relative to the sealing cap. By using the sealing cap, the liquid storage disc can be sealed after sample processing, allowing for complete removal and further reducing the risk of contamination. Furthermore, both the liquid storage disc and the sealing cap are mounted on the guide bracket and can rotate relative to each other without affecting the rotation of the liquid storage disc.
[0021] In one embodiment, the test kit further includes a resilient reset element, one end of which abuts against the sealing cap, and the other end abuts against the inner wall of the kit, and the resilient reset element is located within the upper cavity. A downward force can be indirectly applied to the liquid storage disc using a resilient reset element such as a spring, achieving the purpose of lowering and resetting the liquid storage disc after the pipetting device has taken liquid.
[0022] In one embodiment, the inner wall of the box is further provided with a guide rod for guiding the guide bracket, and the sealing cover is provided with a guide sleeve that cooperates with the guide rod. Both the guide rod and the guide sleeve are located within the upper cavity. The guiding function can be achieved by sleeved over the guide rod.
[0023] In one embodiment, the sealing cap is mounted on the guide bracket and has a working position and a sealed position. When the sealing cap is in the working position, it can rotate relative to the liquid storage turntable. When the sealing cap is in the sealed position, it engages with the liquid storage turntable to seal the liquid tank. The sealing cap has at least two liquid extraction holes, the positions of which correspond to the positions of the pipette tip and the sample inlet, respectively. It is understood that when the sample undergoes pretreatment pipetting or other steps in the cartridge, the pipette tip or external mechanism can extract and add the sample through the extraction holes. After the sample has completed the pretreatment steps, the liquid tank and extraction holes can be misaligned and engaged by rotating the liquid storage turntable, thereby sealing the liquid tank.
[0024] In one embodiment, the test kit further includes a sample tube comprising a snap-fit cap and a tube body. The tube body is fixed inside the sample tube opening by a snap-fit connection. The cap has a sealing top cover, which is detachably connected to the cap by a tight fit. The sample tube is used to hold the pre-treated sample. Through the snap-fit cap and tube body, the sample can be transferred into the tube body after pre-treatment, and the cap can be fastened to the tube body using the sealing top cover. The sealed sample tube can then be transferred to other equipment for testing. At this point, the sample tube and the kit are separated, and both are sealed after separation.
[0025] Since these sample tubes are PCR reaction tubes, and subsequent amplification and detection are required, the PCR reaction tubes can be separated and amplification and fluorescence detection can be performed on a qPCR instrument. Real-time quantitative PCR (qPCR) instruments can simultaneously detect multiple samples (one PCR reaction tube per sample and one test), and the volume is relatively small. This avoids the drawbacks of directly inserting PCR reaction tubes and cartridges into the qPCR instrument, which can lead to incompatibility with the instrument, inaccurate results, and large space requirements, making it unsuitable for multi-sample testing.
[0026] At the same time, there is no limit to the number of sample tubes; the number can be n, which means that one sample can be used to perform joint testing of n test items.
[0027] In one embodiment, the outer wall of the tube is provided with a groove, and a tube sealing ring is provided in the groove, so that the tube and the liquid storage turntable are dynamically sealed.
[0028] In one embodiment, the test kit further includes a cap insertion mechanism, which is fixed to the inner wall of the kit and protrudes towards the liquid storage disc. The cap can be pushed against the sealing cap by the insertion mechanism to lock the cap and the tube body together, thus achieving a seal between the cap and the tube body.
[0029] The present invention also discloses a sample pretreatment device, including the above-mentioned test reagent cartridge and a driving device. The driving device includes a lifting device and a rotating device. The rotating device drives the liquid storage turntable to rotate along its rotation axis, and the lifting device drives the liquid storage turntable to rise and fall.
[0030] The aforementioned sample pretreatment equipment uses a lifting and rotating device to drive the liquid storage turntable to lift and rotate, thereby completing the liquid transfer and promoting the sample pretreatment reaction.
[0031] In one embodiment, the lifting device includes a push rod and a push rod driver. One end of the push rod is mounted on the push rod driver, and the other end abuts against the liquid storage turntable. The push rod driver drives the push rod to retract or extend, thereby moving the liquid storage turntable up and down. The rotating device includes a central shaft of the liquid storage turntable and a drive device for rotating the central shaft. For example, an external device can be used to clamp and rotate the central shaft, causing the liquid storage turntable to perform circular motion.
[0032] In one embodiment, the sample pretreatment device further includes a pipetting drive device. The pipette has a piston rod and an elastic piston inside. The elastic piston has a rod hole that fits tightly with the piston rod. The pipetting drive device is a gripping device for driving the piston rod to move.
[0033] In one embodiment, the sample pretreatment device further includes an incubation and shaking device, a magnetic suction device, and a moving mechanism. The liquid tank includes a reagent tank, a reaction tank, and a waste liquid tank. Both the incubation and shaking device and the magnetic suction device can be moved closer to or away from the reaction tank by the moving mechanism. It is understood that if the sample detection is a molecular diagnostic test, the pretreatment device includes an incubation and shaking device and a magnetic suction device for nucleic acid extraction and purification, and utilizes a conventional moving mechanism to move these devices. When incubation and shaking or magnetic suction is required, the corresponding device is moved closer to or covers the corresponding liquid tank (such as the reaction tank) to complete the pretreatment work.
[0034] In one embodiment, the sample pretreatment device further includes a mechanical gripper for gripping the sample tube. It is understood that the mechanical gripper uses conventional mechanical components and can be configured according to common practice in the art. However, through the design of the mechanical gripper in conjunction with the sample tube, the pretreatment device achieves fully automated operation while also possessing advantages such as small footprint and high flexibility.
[0035] This invention also discloses a sample pretreatment method, which uses the above-mentioned pretreatment equipment and includes the following steps:
[0036] Sample addition: Open the card box cover, control the rotating device to drive the liquid storage turntable to rotate until the liquid tank is aligned with the sample inlet, add the sample to be processed and reagents, and close the card box cover tightly;
[0037] The reaction is as follows: the rotating device is controlled to drive the liquid storage turntable to rotate, so that the position of the pipette tip corresponds to the position of the desired liquid tank. The lifting device is controlled to drive the liquid storage turntable to rise, so that the pipette tip is located in the corresponding liquid tank. The pipette is controlled to draw liquid. The lifting device is controlled to lower the liquid storage turntable. The liquid storage turntable is controlled to rotate to the predetermined position to complete the pipetting and allow the sample to complete the pretreatment reaction in the liquid tank.
[0038] Sampling: Control the pipetting device to aspirate the pretreated sample solution, transfer it into a sample tube, and then remove the sample tube.
[0039] Compared with the prior art, the present invention has the following beneficial effects:
[0040] This invention provides a test reagent cartridge equipped with a cartridge lid. After all sample is added through the sample inlet, the cartridge can be tightly closed. Subsequent sample pretreatment processes are all completed inside the cartridge, ensuring a sealed and contamination-free process. Furthermore, this test reagent cartridge, through the rotation of the liquid storage disc and the cooperation of the sample inlet, eliminates the need for pre-sealed reagents inside the cartridge, facilitating its adaptation for different testing applications.
[0041] By using a rotating liquid storage tray, the axis of the liquid tank is aligned with the axis of the pipette tip. The liquid tank and the pipette tip are all located at the same rotation center of the cartridge. Compared to the method of moving the pipette tip or the method of moving the liquid storage tray, the rotation method saves more space. The method of moving the pipette tip requires storage space after the movement, while the rotation method does not require extra storage space. The cartridge volume can be designed to be smaller and more space-saving.
[0042] Furthermore, the processing, manufacturing, and assembly of this card box do not involve high-difficulty components or equipment, giving it the advantages of low manufacturing and assembly costs and controllable yield rates. Attached Figure Description
[0043] Figure 1 Schematic diagram of the test reagent cartridge structure Figure 1 ;
[0044] Figure 2 Schematic diagram of the test reagent cartridge structure Figure 2 ;
[0045] Figure 3 This is a schematic diagram of the exploded structure of a test reagent cartridge.
[0046] Figure 4 This is a schematic diagram of the external structure of the test reagent cartridge;
[0047] Figure 5 This is a schematic diagram of the liquid storage rotating disk structure;
[0048] Figure 6 This is a schematic diagram of the pipetting device and guide rod structure;
[0049] Figure 7 This is a schematic diagram of the sample tube structure;
[0050] Figure 8 Top view of the liquid storage turntable and sample tube;
[0051] Figure 9 for Figure 8 EE cross-section;
[0052] Figure 10 Top view of the sealing cap;
[0053] Figure 11 This is a cross-sectional view of the sealing cap DD;
[0054] Figure 12 This is a schematic diagram showing the direction of movement of the liquid storage disc inside the test reagent cartridge.
[0055] Wherein: 100, liquid storage turntable; 110, reaction tank; 120, waste liquid tank; 131, first reagent tank; 132, second reagent tank; 133, third reagent tank; 134, fourth reagent tank; 141, first clearance hole; 142, second clearance hole; 143, third clearance hole; 144, fourth clearance hole; 145, fifth clearance hole; 150, sample tube hole; 200, pipetting device; 211, pipette tip No. 1; 212 1. Pipette No. 2; 300. Box body; 310. Sample inlet; 320. Box cover; 330. Top-insertion mechanism; 400. Sample tube; 410. Tube cap; 421. Tube body; 422. Tube body sealing ring; 500. Guide bracket; 510. Turntable sealing ring; 520. Elastic reset component; 530. Limiting component; 540. Guide rod; 600. Sealing cap; 610. Liquid dispensing hole; 620. Guide sleeve; 630. Sealing top cover. Detailed Implementation
[0056] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.
[0057] It should be noted that when a component is said to be "fixed to" another component, it can be directly on the other component or there may be an intervening component. When a component is said to be "mounted" to another component, it can be directly mounted to the other component or there may be an intervening component.
[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.
[0059] Example 1
[0060] A test kit, such as Figure 1-4 It includes: a liquid storage turntable 100, a pipetting device 200, and a housing 300. Depending on the needs, a dedicated sample tube 400 may or may not be provided; in this embodiment, the example is described with a sample tube 400 provided.
[0061] The liquid storage turntable 100 is provided with a plurality of liquid tanks for containing liquid and sample tube holes 150 for containing sample tubes 400. The sample tube holes 150 and the plurality of liquid tanks are arranged around the rotation axis of the liquid storage turntable 100. Specifically, the plurality of liquid tanks and the sample tube holes 150 are arranged in a ring around the axis of the liquid storage turntable 100, and the pipette head and the sample inlet 310 are respectively positioned opposite to the ring.
[0062] In this embodiment, the liquid tank includes reagent tanks (first reagent tank 131, second reagent tank 132, third reagent tank 133, fourth reagent tank 134), reaction tank 110, and waste liquid tank 120. It is understood that the number and function of each type of tank in the specific liquid tank can be set according to the requirements of the specific testing project. For example, for testing projects that generate a large amount of waste liquid, a larger waste liquid tank can be designed to avoid leakage or contamination caused by subsequent processing operations such as shaking the sample. In this embodiment, the test reagent cartridge also includes a guide bracket 500. The liquid storage turntable 100 is installed inside the box body 300 through the guide bracket 500. The turntable sealing ring 510 is sleeved on the outer peripheral edge of the guide bracket 500, so that the guide bracket 500 drives the liquid storage turntable 100 to form a dynamic seal with the box body 300.
[0063] For limiting the downward movement of the liquid storage turntable 100, the inner wall of the box body 300 is also provided with a limiting member 530 for restricting the downward movement of the liquid storage turntable 100. The limiting member 530 is disposed in the lower cavity. The limiting member 530 is used to prevent the liquid storage turntable 100 from falling out of the box body 300 after excessive downward movement. It can be understood that the specific shape and position of the limiting member 530 can be set according to conventional technology in the art. In this embodiment, the limiting member 530 is a limiting plate that is installed around the inner wall of the card box.
[0064] The pipetting device 200 includes a piston-type pipette (such as a piston-type aspiration element similar in structure to a syringe) for aspirating and dispensing liquid, the position of the pipette tip corresponding to the position of the liquid reservoir. It is understood that the piston-type pipette can be a device such as a syringe, and the required accuracy can be adjusted by controlling the tube diameter, i.e., the volume of liquid that can be transferred by moving the piston a certain distance.
[0065] In this embodiment, two pipettes are used, and the liquid storage turntable 100 is also provided with a clearance hole. When one pipette corresponds to the position of the liquid tank, the other pipette corresponds to the position of the clearance hole. It is understood that since the position of the pipettes remains unchanged when the liquid storage turntable 100 moves upward, if there are two pipettes, and one pipette is used while the other is idle, if the idle pipette has no corresponding hole, it will collide with the surface of the liquid storage turntable 100. Therefore, clearance design for the pipettes needs to be considered. The specific location of the clearance hole can be determined by those skilled in the art based on the specific testing process requirements, as well as the number and location of the applicable pipettes. The clearance hole can be a specially designed hole or another reagent tank that does not affect the testing process.
[0066] Based on the requirement of molecular diagnostics for the detection items in this embodiment, and the design of the two pipette tips, as follows: Figure 5-6 As shown, in this embodiment, the holes on the liquid storage turntable 100 are arranged to accommodate the sample solution to be processed and the reaction tank 110 for reaction. Along the circumference of the liquid storage turntable 100, the following are arranged in sequence: reaction tank 110, fourth clearance hole 144, waste liquid tank 120, third clearance hole 143, fourth reagent tank 134, second reagent tank 132, third reagent tank 133, first clearance hole 141, second clearance hole 142, sample tube hole 150, and fifth clearance hole 145.
[0067] The aforementioned liquid storage turntable 100 is installed inside the box body 300 via a guide bracket 500, dividing the internal space of the box body 300 into an upper cavity above the surface of the liquid storage turntable 100 and a lower cavity below the surface of the liquid storage turntable 100. The liquid storage turntable 100 can rotate and move up and down along the inner wall of the box body 300. The pipetting device 200 is fixed on the box body 300, and the pipetting head is located in the upper cavity. The liquid storage turntable 100 and the inner wall of the box body 300 form a sealed space in the upper cavity. The box body 300 is provided with a sample inlet 310 communicating with the upper cavity and a matching card cover 320 for the sample inlet 310.
[0068] To further reduce the risk of contamination, the cartridge in this embodiment also includes a sealing cap 600 for sealing the liquid storage turntable 100. The sealing cap 600 is mounted on the guide bracket 500, and the liquid storage turntable 100 can rotate relative to the sealing cap 600. By using the sealing cap, the liquid storage turntable can be sealed after sample processing, allowing for complete removal and further reducing the risk of contamination. Furthermore, both the liquid storage turntable and the sealing cap are mounted on the guide bracket and can rotate relative to each other without affecting the rotation of the liquid storage turntable.
[0069] In this embodiment, the test reagent cartridge further includes an elastic reset member 520. One end of the elastic reset member 520 abuts against the sealing cap 600, and the other end abuts against the inner wall of the cartridge body. A downward force can be indirectly applied to the liquid storage disc 100 through the elastic reset member 520, such as a spring, to achieve the purpose of lowering and resetting the liquid storage disc after the pipetting device has taken liquid.
[0070] The inner wall of the housing is also provided with a downwardly extending guide rod 540 for guiding the guide bracket 500. The sealing cover 600 is provided with a guide sleeve 620 that cooperates with the guide rod 540. Both the guide rod 540 and the guide sleeve 620 are located in the upper cavity. The guiding function can be achieved by sleeved over the guide rod. The elastic reset member 520 (spring) is also sleeved over the guide sleeve 620.
[0071] Specifically, the sealing cap 600 is mounted on the guide bracket 500 and has a working state position and a sealed state position. When the sealing cap 600 is in the working state position, it can rotate relative to the liquid storage turntable 100. When the sealing cap 600 is in the sealed state position, it engages with the liquid storage turntable 100 to seal the liquid tank. The sealing cap 600 has at least two liquid extraction holes 610, the positions of which correspond to the positions of the pipette head and the sample inlet 310, respectively. It can be understood that when the sample undergoes pretreatment pipetting or other steps in the cartridge, the pipette head or external mechanism can extract and add samples through the liquid extraction holes 610; and after the sample has completed the pretreatment steps, the liquid tank and the liquid extraction holes 610 can be misaligned and engaged by rotating the liquid storage turntable 100, thereby achieving the purpose of sealing the liquid tank.
[0072] In this embodiment, the sample tube 400 includes a snap-fit cap 410 and a tube body 420, such as... Figure 7-11 As shown, the tube body 420 is fixed inside the sample tube hole 150 by a snap-fit connection, and the sealing cap 600 is provided with a sealing top cover 630. The tube cap 410 is detachably connected to the sealing top cover 630 by a tight fit.
[0073] The sample tube 400 is used to hold the pre-treated sample. Through the snap-fit cap 410 and tube body 420, the sample can be transferred into the tube body 420 after pre-treatment. The cap 410 is then fastened to the tube body 420 using the sealing top cover 630 on the sealing cap 600. The sealed sample tube 400 is then transferred to other equipment for testing. At this point, the sample tube 400 is separated from the cartridge, and both are sealed after separation.
[0074] Specifically, the outer wall of the tube body 420 is provided with a groove, and a sealing ring for the tube body 420 is provided in the groove, and the tube body 420 is dynamically sealed with the liquid storage turntable 100.
[0075] To enable the cap 410 to be fastened onto the tube body 420, the test reagent cartridge also includes a cap 410 insertion mechanism 330. This mechanism 330 is fixed to the inner wall of the cartridge body 300 and protrudes towards the liquid storage turntable 100. The insertion mechanism 330 pushes the sealing cap 630, which is mounted on the sealing cap 600, to compress the cap 410 and the tube body 420, thus achieving a seal between the cap 410 and the tube body 420 and enabling fully automated operation.
[0076] Example 2
[0077] A sample pretreatment device for molecular diagnostics employs the reagent cartridge and driving device of Embodiment 1 above. The driving device includes a lifting device and a rotating device. The rotating device drives the liquid storage turntable 100 to rotate along its rotation axis, and the lifting device drives the liquid storage turntable 100 to rise and fall. Figure 12 As shown.
[0078] The above-mentioned sample pretreatment equipment uses a lifting device and a rotating device to drive the liquid storage turntable 100 to lift and rotate, so as to complete the liquid transfer and promote the sample pretreatment reaction.
[0079] Specifically, the lifting device includes a push rod and a push rod driver. One end of the push rod is mounted on the push rod driver, and the other end abuts against the liquid storage turntable 100. The push rod driver drives the push rod to retract or extend, thereby driving the liquid storage turntable 100 to move up and down. The rotating device includes a central shaft of the liquid storage turntable 100 and a driving device for rotating the central shaft. For example, an external device can be used to clamp and rotate the central shaft, causing the liquid storage turntable 100 to perform circular motion.
[0080] To achieve automated operation, the sample pretreatment equipment also includes a pipetting drive device. The pipette has a piston rod and an elastic piston inside. The elastic piston has a rod hole that fits tightly with the piston rod. The pipetting drive device is a gripping device used to drive the piston rod to move.
[0081] For example, the piston rod is aligned with the central axis of pipette #1 in the cartridge and moves downwards along the axis until the piston reaches its lowest point. At this point, the top of the piston rod will be tensioned into the hole on the surface of the piston (which is made of elastic material), achieving integration of the piston rod and piston. This drives the piston rod to move up and down, thus achieving pipetting with pipette #1. Similarly, the gripping device grips the piston rod of pipette #2, integrating the piston rod and piston. This drives the gripping device to move up and down, achieving pipetting with pipette head #212.
[0082] Since the pretreatment equipment in this embodiment is used for molecular diagnostic procedures, it also requires nucleic acid extraction and purification of the sample, such as using magnetic beads for extraction and purification. Therefore, the equipment also includes an incubation and shaking device, a magnetic suction device, and a moving mechanism. Both the incubation and shaking device and the magnetic suction device can be moved closer to or further away from the reaction tank 110 by the moving mechanism. It is understood that if the sample detection is for molecular diagnostic testing, the pretreatment equipment includes an incubation and shaking device and a magnetic suction device for nucleic acid extraction and purification, and utilizes a conventional moving mechanism to move these devices. When incubation and shaking or magnetic suction are required, the corresponding device is moved closer to or covers the corresponding liquid tank (such as the reaction tank 110) to complete the pretreatment work.
[0083] Furthermore, the sample pretreatment device also includes a mechanical gripper for gripping the sample tube 400. It is understood that the mechanical gripper uses conventional mechanical components and can be configured according to common practices in the field. However, through the design of the mechanical gripper in conjunction with the sample tube 400, the pretreatment device achieves fully automated operation while also possessing the advantages of small footprint and high equipment flexibility.
[0084] Example 3
[0085] A method for preprocessing molecular diagnostic test samples, using the preprocessing equipment described in Example 2 above, includes the following steps:
[0086] 1. Adding samples.
[0087] Open the cartridge cover 320, control the rotating device to drive the liquid storage turntable 100 to rotate until the liquid tank is aligned with the sample inlet 310, add the sample to be processed and reagents, and then close the cartridge cover 320 tightly. Specifically, in this embodiment:
[0088] 1. An external mechanism drives the card holder cover 320 to open;
[0089] 2. At this time, the reaction tank 110 on the liquid storage turntable 100 is aligned with the axis of the sample inlet 310. 300uL of RNA capture solution, 20uL of proteinase K and 15uL of magnetic bead suspension are added sequentially by the pipette of the automated instrument outside the cartridge.
[0090] 3. Rotate the liquid storage turntable 100. At this time, the second reagent tank 132 is aligned with the axis of the sample inlet 310. Add 500uL of rinsing solution PWC at once using the automated instrument pipette outside the cartridge.
[0091] 4. Rotate the liquid storage turntable 100. At this time, the third reagent tank 133 is aligned with the axis of the sample inlet 310. Add 1000uL of rinsing solution PWE at once using the automated instrument pipette outside the cartridge.
[0092] 5. Rotate the liquid storage turntable 100. At this time, the fourth reagent tank 134 is aligned with the axis of the sample inlet 310. Add 200uL of RNase-free double-distilled water at once using the automated instrument pipette outside the cartridge.
[0093] 6. Rotate the liquid storage turntable 100. At this time, the reaction tank 110 is aligned with the axis of the sample inlet 310. Add 200uL of sample to the external mechanism (this sample addition process needs to be carried out in a closed environment equipped with a high-efficiency filter in the instrument).
[0094] 7. Screw the card holder cover 320 onto the external mechanism to completely seal the entire card holder.
[0095] II. Pretreatment reaction.
[0096] The rotating device is controlled to drive the liquid storage turntable 100 to rotate, so that the position of the pipette head corresponds to the desired liquid tank position. The lifting device is controlled to drive the liquid storage turntable 100 to rise, so that the pipette head is positioned in the corresponding liquid tank. The pipette device 200 is controlled to draw liquid. The lifting device is controlled to lower the liquid storage turntable 100. The liquid storage turntable 100 is controlled to rotate to a predetermined position, such as... Figure 11 As shown, the liquid transfer is completed, and the sample undergoes the pretreatment reaction in the liquid tank. Specifically, in this embodiment:
[0097] 1. After the sample is added to reaction tank 110, the reaction tank 110 contains: sample + 300uL RNA capture solution + 20uL proteinase K + 15uL magnetic bead suspension;
[0098] 2. Add an incubation and shaking device to the reaction tank 110 to incubate and shake the liquid in the reaction tank 110, complete the lysis of the sample, release the nucleic acid, and shake it to allow the nucleic acid to be adsorbed onto the magnetic beads.
[0099] 3. Add a magnetic ring to the reaction tank 110 to allow the magnetic beads with adsorbed nucleic acid to adhere to the side wall of the reaction tank 110; 4. Rotate the liquid storage turntable 100 to align the axis of pipette 211 with the reaction tank 110, then push the entire liquid storage turntable 100 upwards, pushing the tip of pipette 211 to a position with only a small gap between it and the bottom of the tank of the reaction tank 110, driving the piston of pipette 211 upwards to aspirate the waste liquid from the reaction tank 110 except for the nucleic acid adsorbed on the side wall; (at this time, pipette 212 is at the fourth clearance hole 144);
[0100] 5. The drive push rod retracts, and the spring against the guide bracket 500 pushes the liquid storage turntable 100 to move downward as a whole until the liquid storage turntable 100 reaches the limit piece 530, completing the downward reset of the liquid storage turntable 100.
[0101] 6. Rotate the liquid storage turntable 100 to align the waste liquid tank 120 with the axis of pipette head 211. Then, push the entire liquid storage turntable 100 upwards, pushing the tip of pipette head 211 to a position with a gap between it and the bottom of the hole in the waste liquid tank 120. Drive the piston of pipette head 211 downwards to discharge the waste liquid in pipette head 211 into the waste liquid tank 120. Retract the push rod, which, against the spring of the guide bracket 500, pushes the entire liquid storage turntable 100 downwards until it reaches the limit member 530, completing the downward reset of the liquid storage turntable 100. (At this time, pipette head 212 is in position 4# hole. 4# hole contains double-distilled water, which will not affect the cleanliness of pipette head 212.)
[0102] 7. Similar to steps 4 and 5, rotate the liquid storage turntable 100 to align the second reagent tank 132 with the axis of pipette 211, and draw 500 μL of rinsing solution PWC; (at this time, pipette 212 is in the first clearance hole 141)
[0103] 8. Similar to step 6, rotate the liquid storage turntable 100 to align the reaction tank 110 with the axis of pipette 1 211, and discharge the PWC in pipette 1 211 into the reaction tank 110; (at this time, pipette 2 212 is in the fourth clearance hole 144); drive the oscillation incubation and magnetic attraction, then absorb the waste liquid, align the waste liquid tank 120 with the axis of pipette 1 211, and discharge the waste liquid in pipette 1 211 into the waste liquid tank 120; (at this time, pipette 2 212 is in the fourth reagent tank 134, which contains double-distilled water and will not affect the cleanliness of pipette 2 212); drive the push rod to retract, and push the liquid storage turntable 100 downward against the spring of the guide bracket 500 until the liquid storage turntable 100 reaches the limit member 530, completing the downward reset of the liquid storage turntable 100;
[0104] 9. Similar to step 7, rotate the liquid storage turntable 100 to align the No. 3 hole with the axis of the No. 1 pipette 211, and draw 500uL of rinsing solution PWE; (at this time, the No. 2 pipette 212 is in the second clearance hole 142);
[0105] 10. Repeat step 8; transfer the rinsed PWE to waste liquid tank 120;
[0106] 11. Repeat steps 9 and 10;
[0107] 12. Similar to step 7, rotate the liquid storage turntable 100 to align the fourth reagent tank 134 with the axis of pipette 211, and draw 100 μL of RNase-free double-distilled water into reaction well #1; (at this time, pipette 212 is in the third pipette tank, because the third pipette tank has been empty after being drawn in steps 9, 10, and 11, and will not affect the cleanliness of pipette 212).
[0108] 13. Repeat step 8; transfer the RNase-free double-distilled water to waste liquid tank 120;
[0109] 14. Similar to step 7, rotate the liquid storage turntable 100 to align the fourth reagent tank 134 with the axis of the first pipette 211, and draw 100 μL of RNase-free double-distilled water into the reaction tank 110; the nucleic acid dissolves in the double-distilled water, drive the oscillation to mix, and reset the liquid storage turntable 100.
[0110] 15. Add a magnetic ring at the position of reaction tank 110, so that the magnetic beads can be attracted to the side wall of reaction tank 110;
[0111] 16. Similar to step 7, rotate the liquid storage turntable 100 to align the reaction tank 110 with the axis of pipette 212, and aspirate 5 μL of purified nucleic acid (keeping it in magnetic adsorption state); (at this time, pipette 211 is in the fifth clearance hole 145).
[0112] 17. Similar to step 8, rotate the liquid storage turntable 100 to align the sample tube well 150 with the axis of pipette tip 212, and discharge the nucleic acid in pipette tip 212 into sample tube 400 in sample tube well 150; (at this time, pipette tip 211 is in well 2).
[0113] 18. Transfer the purified nucleic acid to sample tube 400 (the reaction tube contains PCR reaction solution).
[0114] III. Sampling.
[0115] The pipetting device 200 is controlled to aspirate the pretreated sample solution and transfer it into the sample tube 400. The sample tube 400 is then removed. Figure 12 As shown below. Specifically:
[0116] 1. Rotate the liquid storage turntable 100 to align the sample tube orifice 150 with the axis of the insertion mechanism 330. Push the entire liquid storage turntable 100 upwards. Simultaneously, control the mechanical gripper to grab the tube body 420 (PCR reaction tube) of the sample tube 400 and push it upwards along with the liquid storage turntable 100. In a synchronized manner, push the liquid storage turntable 100 so that the sealing cap 600 covers the turntable 100. At this point, the vertical height of the liquid storage turntable 100 is higher than the position when the liquid was aspirated. Continue pushing upwards until the tube cap 410 covers the tube body 420 (i.e., the PCR reaction tube), completing the sealing of the PCR tube. Continue pushing the liquid storage turntable 100 upwards, but the mechanical gripper grabbing the sample tube 400 no longer follows the liquid storage turntable 100 upwards. At this point, the sample tube 400 is capped and detached from the cartridge body. The inside of the cartridge remains a closed structure, and the PCR tube is also a fully closed structure, thus achieving a seal between the two.
[0117] 2. The external driver picks up the card holder and discards it;
[0118] 3. The mechanical gripper picks up the sample tube 400 (PCR reaction tube) and places it onto the qPCR instrument for amplification and detection. This completes the PCR extraction, amplification, and detection process.
[0119] The above-mentioned molecular diagnostic sample pretreatment process is a fully sealed process. The sealing is achieved through two steps: First, the entire liquid transfer process of nucleic acid extraction and system construction is completed inside the cartridge, thus achieving a seal; Second, after the PCR reaction tube is separated from the cartridge, the inside of the cartridge remains a closed structure, and the PCR tube is also a fully sealed structure, thus achieving a seal.
[0120] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0121] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.
Claims
1. A test reagent cartridge, characterized in that, include: A liquid storage turntable, wherein the liquid storage turntable is provided with a plurality of liquid tanks for containing liquid and sample tube holes for containing sample tubes, the sample tube holes and the plurality of liquid tanks being arranged around the rotation axis of the liquid storage turntable; A pipetting apparatus, comprising a pipette for aspirating and dispensing liquid, wherein the position of the pipette tip corresponds to the position of the liquid tank, and the pipette is a piston-type pipette; as well as The container has a liquid storage turntable installed inside, dividing the internal space into an upper cavity above the turntable surface and a lower cavity below the turntable surface. The turntable can rotate and move up and down along the inner wall of the container. The pipetting device is fixed to the container, with the pipetting head located in the upper cavity. The turntable and the inner wall of the container form a sealed space in the upper cavity. The container has a sample inlet communicating with the upper cavity and a matching cap for the sample inlet. A plurality of liquid tanks and sample tube holes are arranged in a ring around the axis of the liquid storage turntable. The pipette head and the sample inlet are both positioned relative to the ring. A turntable sealing ring is provided between the liquid storage turntable and the inner wall of the box, and the liquid storage turntable and the box form a dynamic seal. The test reagent cartridge also includes a guide bracket, an elastic reset component, and a sealing cap for sealing the liquid storage disc. The liquid storage disc is mounted in the cartridge body via the guide bracket. The disc sealing ring is fitted around the outer periphery of the guide bracket. The sealing cap is mounted on the guide bracket. The liquid storage disc can rotate relative to the sealing cap. One end of the elastic reset component abuts against the sealing cap, and the other end abuts against the inner wall of the cartridge body. The elastic reset component is located in the upper cavity. The sealing cap is mounted on the guide bracket and has a working position and a sealed position. When the sealing cap is in the working position, the sealing cap and the liquid storage disc can rotate relative to each other. When the sealing cap is in the sealed position, the sealing cap engages with the liquid storage disc and seals the liquid tank. The sealing cap has at least two liquid dispensing holes, the positions of which correspond to the positions of the pipette head and the sample inlet, respectively.
2. The test reagent cartridge according to claim 1, characterized in that, The liquid tank includes a reagent tank, a reaction tank, and a waste liquid tank.
3. The test reagent cartridge according to claim 2, characterized in that, There are two pipettes, and the liquid storage turntable is also provided with a clearance hole. When one of the pipettes is aligned with the position of the liquid tank, the other pipette is aligned with the position of the clearance hole.
4. The test reagent cartridge according to claim 1, characterized in that, The inner wall of the box is also provided with a limiting member for restricting the downward movement of the liquid storage turntable, and the limiting member is located in the lower cavity.
5. The test reagent cartridge according to claim 1, characterized in that, The inner wall of the box is also provided with a guide rod for guiding the guide bracket, and the sealing cover is provided with a guide sleeve that cooperates with the guide rod. Both the guide rod and the guide sleeve are located in the upper cavity.
6. The test reagent cartridge according to claim 1, characterized in that, It also includes a sample tube, which includes a cap and a body that fit together with a snap-fit mechanism. The body is fixed inside the sample tube hole by a snap-fit connection. The cap has a sealing top cover, and the cap is detachably connected to the sealing top cover by a tight fit.
7. The test reagent cartridge according to claim 6, characterized in that, The outer wall of the tube is provided with a groove, and a tube sealing ring is provided in the groove, so that the tube and the liquid storage turntable are dynamically sealed.
8. The test reagent cartridge according to claim 6, characterized in that, It also includes a cap insertion mechanism, which is fixed to the inner wall of the box and protrudes toward the liquid storage turntable.
9. A sample pretreatment device, characterized in that, The invention includes the test reagent cartridge according to any one of claims 1-8, and a driving device, the driving device comprising a lifting device and a rotating device, the rotating device driving the liquid storage turntable to rotate along its rotation axis, and the lifting device driving the liquid storage turntable to rise and fall.
10. The sample pretreatment device according to claim 9, characterized in that, The lifting device includes a push rod and a push rod driver. One end of the push rod is mounted on the push rod driver, and the other end abuts against the liquid storage turntable. The push rod driver drives the push rod to retract or extend, thereby driving the liquid storage turntable to move up and down. The rotating device includes a central shaft of the liquid storage turntable and a driving device for driving the central shaft to rotate.
11. The sample pretreatment device according to claim 9, characterized in that, It also includes a pipetting drive device, wherein the pipette is provided with a piston rod and an elastic piston, the elastic piston is provided with a rod hole that fits tightly with the piston rod, and the pipetting drive device is a gripping device for driving the piston rod to move.
12. The sample pretreatment device according to claim 9, characterized in that, It also includes an incubation and shaking device, a magnetic attraction device, and a moving mechanism. The liquid tank includes a reagent tank, a reaction tank, and a waste liquid tank. The incubation and shaking device and the magnetic attraction device can move closer to or further away from the reaction tank under the action of the moving mechanism.
13. The sample pretreatment device according to claim 9, characterized in that, It also includes a mechanical gripper for gripping the sample tube.
14. A sample pretreatment method, characterized in that, The pretreatment apparatus according to any one of claims 9-13 includes the following steps: Sample addition: Open the card box cover, control the rotating device to drive the liquid storage turntable to rotate until the liquid tank is aligned with the sample inlet, add the sample to be processed and reagents, and close the card box cover tightly; The reaction is as follows: the rotating device is controlled to drive the liquid storage turntable to rotate, so that the position of the pipette tip corresponds to the position of the desired liquid tank. The lifting device is controlled to drive the liquid storage turntable to rise, so that the pipette tip is located in the corresponding liquid tank. The pipette is controlled to draw liquid. The lifting device is controlled to lower the liquid storage turntable. The liquid storage turntable is controlled to rotate to the predetermined position to complete the pipetting and allow the sample to complete the pretreatment reaction in the liquid tank. Sampling: Control the pipetting device to aspirate the pretreated sample solution, transfer it into a sample tube, and then remove the sample tube.