Container for PCR (Polymerase Chain Reaction) detection reagent and PCR pretreatment system

Abstract

The utility model discloses a PCR (Polymerase Chain Reaction) detection reagent container and a PCR pretreatment system. The PCR detection reagent container comprises a box body, a flexible sealing film, a magnetic bar sleeve, a pipettor and a thin sealing film, the box body comprises a plurality of grooves and at least one test tube, and the thin sealing films are arranged at openings of the grooves and the test tubes, so that the grooves or the test tubes are individually sealed. And the flexible sealing film, the box body, the magnetic bar sleeve and the pipettor jointly define a closed space. The PCR pretreatment system comprises the PCR detection reagent container, and a magnetic bar, a suction push rod and a mechanical arm which are matched with the PCR detection reagent container for use. The magnetic bar sleeve and the pipettor move in the X-axis direction, the Y-axis direction and the Z-axis direction in a sealed environment, so that the pollution risk caused by the fact that a detection reagent is exposed in the external environment is avoided, and the accuracy and convenience of detection are guaranteed.

Description

Technical Field

[0001] The present application belongs to the field of biological detection technology, and specifically relates to a container for PCR detection reagents and a PCR pre-treatment system. Background Art

[0002] The automatic pre-treatment method of PCR (Polymerase Chain Reaction) generally uses an automatic nucleic acid extraction system or an automatic nucleic acid extraction workstation to complete the extraction of nucleic acids such as RNA (RiboNucleic Acid) and DNA (DeoxyriboNucleic Acid). When extracting DNA, the open environment may lead to mutual contamination between samples, resulting in inaccurate diagnosis or even misdiagnosis.

[0003] In order to solve the above problems, there is a POCT (point-of-care testing) test kit used with a needle tube, whose elastic sealing component can rebound and seal the pinhole after being punctured by a needle, thereby meeting the requirements of the container being sealed when taking or discharging liquid in each step of PCR pretreatment to prevent the internal and external gases from being connected. However, due to the need for frequent pipetting during the pretreatment process, the needle tube is inevitably exposed to the external environment during the process of entering and exiting the POCT test kit, and there is still a risk of contamination. At the same time, since only the needle tube can enter and exit the POCT test kit through the elastic sealing component, the magnetic rod cannot be applied to this POCT test kit, so only the needle tube can be used to pipette the magnetic bead solution, so that the magnetic beads are easily retained in the needle tube; and only the side suction method can be used to separate the magnetic beads and the solution, resulting in the magnetic beads being easily drawn away by the needle tube, which ultimately affects the measurement accuracy. Furthermore, the pipetting volume requirements of different steps in the pretreatment process vary, the steps are numerous, and the use of existing technology makes the operation inconvenient.

[0004] Therefore, there is an urgent need to propose a container for PCR detection reagents and a PCR pretreatment system that can be used for POCT and can complete PCR pretreatment and PCR detection completely in a sealed environment. Utility Model Content

[0005] In view of the above-mentioned shortcomings or deficiencies of the prior art, the technical problem to be solved by the present application is to provide a container for PCR detection reagents and a PCR pre-treatment system.

[0006] To solve the above technical problems, this application is implemented through the following technical solutions:

[0007] The present application proposes a container for PCR detection reagents, comprising: a box body, a flexible sealing film, a magnetic rod sleeve, a pipette and a thin sealing film; the box body comprises a reagent accommodating portion and an amplification tube accommodating portion, the reagent accommodating portion is provided with a plurality of grooves, the amplification tube accommodating portion is provided with at least one test tube, the thin sealing film is arranged at the openings of the grooves and the test tubes, so that the grooves or the test tubes are individually sealed; the flexible sealing film is fixedly and sealedly connected to the outer peripheries of the magnetic rod sleeve and the pipette; the flexible sealing film is fixedly covered on the top surface of the box body, and together with the box body, the magnetic rod sleeve and the pipette, forms a closed space of the container.

[0008] Optionally, the above-mentioned container of PCR detection reagent further comprises: a cover frame, wherein the cover frame is fixedly connected to the flexible sealing film and the box body, so that the flexible sealing film and the box body are sealed;

[0009] Optionally, in the container of the above-mentioned PCR detection reagent, the flexible sealing film is an airtight, watertight, elastic film with good elongation at break performance; the airtightness and watertightness enable the flexible sealing film to seal the closed space of the container; the elasticity enables the flexible sealing film to rebound and reseal the needle hole after being punctured by a needle, and to return to its original state after being stretched. In the stretched state, the elongation at break performance of the flexible sealing film should meet the requirements that the magnetic rod sleeve and the pipette are withdrawn from their respective accommodating grooves separately or together to complete the vertical stroke, and / or the horizontal stroke is completed without breaking, so that the closed space of the container always remains in a closed state; in the unstretched state, the surface of the flexible sealing film is flat.

[0010] Optionally, in the above-mentioned container of PCR detection reagent, the flexible sealing film is made of TPV, TPE, TPR, silicone or latex.

[0011] Optionally, in the above-mentioned container of PCR detection reagent, the flexible sealing film is provided with at least one piercing identification area to facilitate accurate insertion of the needle into the corresponding groove or test tube.

[0012] Optionally, in the above-mentioned container of PCR detection reagent, the box body or the flexible sealing film is provided with an identification code for enabling the automatic nucleic acid extraction system or workstation to accurately identify and match the corresponding container of PCR detection reagent.

[0013] Optionally, in the above-mentioned container of PCR detection reagent, the cover frame, the flexible sealing film, the magnetic rod sleeve and the pipette are integrally formed.

[0014] Optionally, in the above-mentioned container of PCR detection reagent, the magnetic rod sleeve is a semi-enclosed tubular body with only the top end open, for accommodating the magnetic rod; the pipette is a cavity structure with top and bottom ends open, and a first piston that can be connected to an external push rod is arranged in the cavity, and the first piston seals the liquid holding area of ​​the pipette from the top.

[0015] Optionally, in the above-mentioned container of PCR detection reagent, the bottom end of the magnetic rod sleeve and / or the pipette has a pointed tip to facilitate cutting the thin sealing film.

[0016] Optionally, a convex annular structure is circumferentially arranged in the middle of the outer wall of the magnetic rod sleeve, and the convex annular structure protrudes circumferentially from the outer wall of the magnetic rod sleeve below it, so as to further expand the opening of the thin sealing film and prevent the magnetic beads from hitting the thin sealing film and falling off when the magnetic rod sleeve exits the groove.

[0017] Optionally, in the above-mentioned container of PCR detection reagent, the box body further includes: a magnetic rod cover receiving groove and a pipette receiving groove, the magnetic rod cover receiving groove is used to place the magnetic rod cover, and the pipette receiving groove is used to place the pipette.

[0018] Optionally, in the above-mentioned container of PCR detection reagent, the reagent containing part and the amplification tube containing part are respectively located on both sides of the box body; and the magnetic rod sleeve containing groove is arranged in the reagent containing part.

[0019] Optionally, in the above-mentioned container for PCR detection reagents, the reagent containing part, the amplification tube containing part, the magnetic rod sleeve containing groove, the pipette containing groove and the wall of the box body are an integrated structure, or are a separate structure but are fixedly sealed and connected by means of snap-fitting or the like.

[0020] Optionally, in the above-mentioned container of PCR detection reagent, the thin sealing film can be one or more sheets, and is made of aluminum-plastic material.

[0021] Optionally, the above-mentioned container of PCR detection reagent further includes: a needle tube; the needle tube includes a needle tube body, a steel needle, and a second piston.

[0022] Optionally, in the above container of PCR detection reagent, the needle tube further comprises: a telescopic needle protection sleeve, the steel needle is wrapped by the telescopic needle protection sleeve, one end of the telescopic needle protection sleeve is fixed to the needle tube body, and the other end is a free end. When the steel needle pierces the flexible sealing film, the free end of the telescopic needle protection sleeve shrinks toward the fixed end due to being blocked by the flexible sealing film; when the steel needle withdraws, the telescopic needle protection sleeve is reset.

[0023] Optionally, the above-mentioned container of PCR detection reagent further includes: a syringe accommodating groove; the syringe accommodating groove is arranged beside the box body outside the closed space of the container, for placing the syringe.

[0024] Optionally, in the above-mentioned container of PCR detection reagent, at least one of the grooves stores paraffin oil. Before PCR pre-amplification or amplification, the pipette moves the paraffin oil to a test tube containing a nucleic acid sample solution or a pre-amplification groove to prevent the solution from evaporating and ensure the accuracy of the amplification reaction results; the paraffin oil can also be moved to the elution groove of the residual nucleic acid sample solution to further achieve a sealing effect and prevent the escape of nucleic acid.

[0025] The present application also proposes a PCR pretreatment system, which includes the container of the aforementioned PCR detection reagent, and also includes: a magnetic rod, a first suction push rod, a second suction push rod, and at least two mechanical arms; the magnetic rod has a magnetic attraction and can be placed in a magnetic rod sleeve; the first suction push rod and the second suction push rod have a connecting portion that is convenient for connecting and disconnecting with the piston, the first suction push rod can be connected to the first piston of the pipette, and the second suction push rod can be connected to the second piston of the syringe; the mechanical arm can control the movement of the magnetic rod sleeve, the pipette, the magnetic rod, the syringe and / or the suction push rod.

[0026] Optionally, in the above-mentioned PCR pre-treatment system, the first suction push rod and the second suction push rod are the same component, and the component can be connected to the first piston and the second piston respectively.

[0027] Optionally, in the above-mentioned PCR pretreatment system, the magnetic rod and the first suction push rod are the same component, that is, the first suction push rod has magnetic attraction, and can be used as a magnetic rod, and can also be connected to the first piston to control the suction or pushing action of the pipette.

[0028] Optionally, in the above-mentioned PCR pretreatment system, the magnetic rod, the first suction push rod, and the second suction push rod are the same component, that is, the component has magnetic attraction, and can be used as a magnetic rod, and can also be respectively connected to the first piston and the second piston to respectively control the suction or pushing action of the pipette and the syringe.

[0029] Optionally, in the above-mentioned PCR pre-treatment system, when the magnetic bar is erected, the magnetic poles are respectively located on the left and right sides or the front and back sides of the magnetic bar.

[0030] Compared with the prior art, this application has the following technical effects:

[0031] The flexible sealing film, box body, magnetic rod cover and pipette of the present application together constitute a closed environment. The magnetic rod cover and the pipette complete the operation in the sealed environment. Almost all pre-treatment steps and PCR amplification steps can be completed in the closed environment of the container, avoiding the risk of contamination caused by exposure to the external environment and ensuring the accuracy of the detection.

[0032] In the prior art, a needle is used to pipette the magnetic bead solution, which makes it easy for the magnetic beads to remain in the needle; and after the elution is completed, the side suction method is used to separate the magnetic beads and the solution, which causes the magnetic beads to be easily drawn away by the needle, ultimately affecting the measurement accuracy. The use of the magnetic bar method can avoid the above problems, but the use of the magnetic bar in the prior art must be completed in an open space, and it is difficult to avoid the risk of contamination. The matching arrangement of the magnetic bar, magnetic bar sleeve, and flexible sealing film in the present application makes it possible to use the magnetic bar method to obtain and separate magnetic beads in a closed environment without frequent pipetting. The pipette can be used only for small volume pipetting operations such as moving the DNA sample solution to the amplification test tube, which is convenient and simple to operate, avoiding and ensuring the detection accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Other features, objects and advantages of the present application will become more apparent by reading the detailed description of non-limiting embodiments made with reference to the following drawings:

[0034] Figure 1 : A schematic diagram of the structure of an embodiment of the present application Figure 1 ;

[0035] Figure 2 : A schematic diagram of the structure of an embodiment of the present application Figure 2 ;

[0036] Figure 3 :like Figure 2 A front view of the structure shown;

[0037] Figure 4 :like Figure 2 Top view of the structure shown Figure 1 ;

[0038] Figure 5 :like Figure 2 Top view of the structure shown Figure 2 ;

[0039] Figure 6 :like Figure 2 Cross-section of the structure shown Figure 1 ;

[0040] Figure 7 :like Figure 2 Cross-section of the structure shown Figure 2 ;

[0041] Figure 8 : Schematic diagram of the combination of a cover frame, a magnetic rod sleeve, a flexible sealing film, and a pipette in one embodiment of the present application;

[0042] Fig. 9 : A front view of the combination of a cover frame, a magnetic rod sleeve, a flexible sealing film, and a pipette in one embodiment of the present application;

[0043] Fig.10: A schematic diagram of an amplification tube accommodating portion in an embodiment of the present application;

[0044] Fig.11 : A schematic diagram of a pipette application in one embodiment of the present application;

[0045] Fig.12 : Schematic diagram of a magnetic rod sleeve in one embodiment of the present application;

[0046] Fig.13 : A schematic structural diagram of another embodiment of the present application;

[0047] In the figure: box body 1, reagent accommodating part 101, magnetic rod sleeve accommodating groove 102, pipette accommodating groove 103, amplification tube accommodating part 104, first groove 105a, second groove 105b, third groove 105c, fourth groove 105d, fifth groove 105e, sixth groove 105f, test tube 106, flexible sealing film 2, piercing identification area 201, magnetic rod sleeve 3, tip 301, annular convex structure 302, pipette 4, first piston 401, first plug-in part 402, tip 403, liquid accommodating area 404, cover frame 5, needle tube accommodating groove 6, needle tube 7, needle tube body 701, telescopic needle protection sleeve 702, second piston 703, second plug-in part 704, steel needle 705. DETAILED DESCRIPTION

[0048] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0049] like Figures 1 to 11 As shown, one of the embodiments of the present application is a container for PCR detection reagents, comprising: a box body 1, a flexible sealing film 2, a magnetic rod sleeve 3, a pipette 4, and a thin sealing film (not shown in the figure); the box body 1 includes a reagent holding part 101 and an amplification tube holding part 104. The reagent holding part 101 is provided with a plurality of grooves 105 for holding PCR pre-treatment reagents. In this embodiment, the PCR pre-treatment reagents include but are not limited to lysate, magnetic beads, washing solution, eluent, paraffin oil or pre-amplification reagents, etc.; the amplification tube holding part 104 is provided with at least one test tube 106 for holding PCR reagents. The thin sealing film covers the reagent holding part 101 and the amplification tube holding part 104, so that each groove 105 and each test tube 106 are independently sealed to prevent the liquid or powder in the groove 105 and the test tube 106 from overflowing during transportation or movement, and the reagents and powders flow and mix with each other.

[0050] The flexible sealing film 2 covers the top surface of the box body; the flexible sealing film 2, the box body 1, the magnetic rod sleeve 3 and the pipette 4 together form a closed container space, so that the magnetic rod sleeve 3 and the pipette 4 are always in a closed environment to complete the operation. Fig.11 as shown, avoiding the risk of contamination caused by exposure to the external environment.

[0051] Optionally, the box body 1 further comprises: a magnetic rod sleeve accommodating groove 102 and a pipette accommodating groove 103 , wherein the magnetic rod sleeve accommodating groove 102 is used to place the magnetic rod sleeve 3 ; and the pipette accommodating groove 103 is used to place the pipette 4 .

[0052] Specifically, the flexible sealing film 2 and the upper side wall of the box body 1 can be fixedly sealed and connected by gluing or by pressing or buckling with a fixing member.

[0053] Optionally, the flexible sealing film 2 is made of TPV (Thermoplastic Vulcanizate), TPE (Thermoplastic Elastomer), TPR (Thermo-Plastic-Rubber), silicone and / or latex to ensure that the flexible sealing film 2 is an airtight, watertight, elastic and elongation-at-break film. Of course, the materials here are not exhaustive, and those skilled in the art are motivated to replace them with other materials according to technical ideas.

[0054] Specifically, after being pierced by a sharp structure such as a needle, the flexible sealing film 2 can rebound by itself and reseal the needle hole.

[0055] Preferably, the elongation at break performance of the flexible sealing film 2 satisfies the requirement that the flexible sealing film 2 will not be stretched and ruptured when the magnetic rod sleeve 3 and the pipette 4 are taken out and complete their respective operation strokes separately or simultaneously, thereby ensuring that the operation process is always in a closed space.

[0056] In this embodiment, the flexible sealing film 2 is made of TPE material.

[0057] Optionally, the container of the PCR detection reagent further comprises: a cover frame 5, the cover frame 5 is fixedly and sealedly connected to the flexible sealing film 2 and the box body 1, so that the flexible sealing film and the box body are sealed. The cover frame 5 and the flexible sealing film 2 can be integrally formed by two-color injection molding or the like, or the cover frame 5 and the flexible sealing film 2 can be connected by gluing, snapping or the like, such as Figure 8 shown.

[0058] The cover frame 5 is snap-fitted with the box body 1, and the flexible sealing film 2 plays a sealing role at the snap-fitting portion between the cover frame 5 and the box body 1. Alternatively, in other embodiments, a sealing ring is provided at the connection between the cover frame 5 and the box body 1.

[0059] Optionally, the reagent containing part 101, the magnetic rod sleeve containing groove 102, the pipette containing groove 103, and the amplification tube containing part 104 are integrally arranged with the box body 1, such as Fig.13 Alternatively, the amplification tube accommodating portion 104 is connected to the box body 1 by snapping. In this embodiment, the amplification tube accommodating portion 104 is snapped to the box body 1 to simplify the design process of the box body 1.

[0060] Optionally, when the amplification tube accommodating portion 104 is snap-fitted to the box body 1 , a sealing gasket is provided at the connection between the box body 1 and the amplification tube accommodating portion 104 to ensure the sealing of the enclosed space.

[0061] Optionally, as in the present embodiment, when the number of the grooves and test tubes is large, the grooves and test tubes are distributed in strips on the box body 1, and the magnetic rod sleeve receiving groove 102 is set in the reagent receiving part 101, so that the magnetic rod sleeve 3 can complete the operation in the smallest possible range, which can reduce the stretching amplitude of the flexible sealing film 2; the pipette receiving groove 103 is set in the reagent receiving part 101, the receiving part 104, or the junction of the reagent receiving part 101 and the amplification tube receiving part 104, so that the pipette 4 can complete the operation in the smallest possible range, which can reduce the stretching amplitude of the flexible sealing film 2, and at the same time can also reduce the operation stroke and improve efficiency. When the number of the grooves and test tubes is small, the pipette receiving groove 103 may be set in the reagent receiving part 101 or the amplification tube receiving part, and the same technical effect can still be achieved.

[0062] Optionally, the thin sealing film is one or more sheets and is made of aluminum-plastic material.

[0063] Specifically, the top end of the magnetic rod sleeve 3 is open and the bottom end is closed. The magnetic rod sleeve 3 can be placed in the magnetic rod sleeve accommodating groove 102, and the outer wall of the part higher than the magnetic rod sleeve accommodating groove 102 is fixedly sealed and connected to the flexible sealing film 2; the bottom end of the magnetic rod sleeve 3 is provided with a pointed end 301 to easily scratch the thin sealing film.

[0064] Alternatively, if Fig.12As shown, a convex ring structure 302 is circumferentially arranged in the middle of the outer wall of the magnetic rod sleeve, and the convex ring structure 302 protrudes circumferentially from the outer wall of the magnetic rod sleeve below it. The convex ring structure 302 allows the magnetic rod sleeve 3 to further expand the opening of the thin sealing film when it continues to be placed in the groove after piercing the thin sealing film, thereby preventing the magnetic beads adsorbed on the lower part of the magnetic rod sleeve 3 from hitting the thin sealing film when the magnetic rod sleeve 3 withdraws from the groove to transfer the magnetic beads, causing the magnetic beads to be scratched or dropped, thereby affecting the detection accuracy.

[0065] Optionally, the connection between the magnetic rod sleeve 3 and the flexible sealing film 2 can be integrally formed by two-color injection molding or the like to form a fixed and sealed connection. The height of the magnetic rod sleeve 3 is greater than the depth of the magnetic rod sleeve accommodating groove 102 .

[0066] Specifically, the pipette 4 is a cavity structure with openings at the top and bottom, and a first piston 401 is built in the middle of the cavity; the first piston 401 is provided with a first plug-in portion 402 for externally connecting a push rod; the pipette 4 can be placed in the pipette accommodating groove 103, and the outer wall of the portion higher than the pipette accommodating groove 103 is fixedly sealed and connected with the flexible sealing film 2; the bottom end of the pipette 4 has a tip 403 to easily cut the thin sealing film. The first piston 401 seals the top of the liquid receiving area 404 of the pipette 4.

[0067] Optionally, the connection between the pipette 4 and the flexible sealing membrane 2 can be integrally formed by two-color injection molding or the like to form a fixed and sealed connection. The height of the pipette 4 is greater than the depth of the pipette accommodating groove 103. Optionally, the pipette can be equipped with a matching suction push rod.

[0068] Preferably, the position where the flexible sealing film 2 is fixedly sealed with the magnetic rod sleeve 3, the position where the flexible sealing film 2 is fixedly sealed with the pipette 4, and the position where the flexible sealing film 2 is fixedly sealed with the cover frame 5 are on the same horizontal plane, so that the flexible sealing film 2 is substantially in a horizontal state in an unstretched state, and the surface is flat. In different embodiments, as known to those skilled in the art, the flexible sealing film is also allowed to be in a plane tilted state, which does not affect the technical effect achieved by the flexible sealing film 2.

[0069] Alternatively, if Fig. 9 As shown, the flexible sealing film 2 is provided with at least one puncture identification area 201 to facilitate accurate insertion of the needle tube into the corresponding groove 105; preferably, the identification area can be marked to facilitate identification of the puncture position; the thickness of the flexible sealing film 2 in the identification area is increased so that the puncture identification area can better rebound and seal after being punctured by the needle.

[0070] Optionally, the box body 1 or the flexible sealing film 2 is provided with an identification code for enabling an automatic nucleic acid extraction system or a workstation to accurately identify the container of the PCR detection reagent to avoid errors or omissions.

[0071] Optionally, the identification code includes but is not limited to a QR code or a bar code, and can be set on the box body 1 or the flexible sealing film 2 by spraying or pasting.

[0072] Optionally, the container of PCR detection reagent also includes: a needle tube accommodating groove 6, which is arranged in an open space outside the enclosed space covered by the flexible sealing film and connected to the box body 1. The needle tube 7 is arranged in the needle tube accommodating groove 6, and includes: a needle tube body 701, a steel needle 705, a telescopic needle protection sleeve 702, and a second piston 703. The steel needle 705 is wrapped by the telescopic needle protection sleeve 702, one end of the telescopic needle protection sleeve 702 is fixed to the needle tube body 701, and the other end is a free end. When the needle tube 7 is pipetting, the needle of the steel needle 705 pierces the flexible sealing film 2, and the free end of the telescopic needle protection sleeve 702 is blocked by the flexible sealing film 2 and shrinks. When the steel needle 705 is withdrawn, the telescopic needle protection sleeve 702 is reset and re-covered on the needle, thereby effectively preventing external contamination. When the container of the PCR detection reagent includes a needle tube 7 and a needle tube receiving groove 6, there is no need to equip the PCR pre-treatment system with an external sample injection needle tube, so that the container of the PCR detection reagent achieves a high degree of integration of the functional set. It is known to those skilled in the art that in another embodiment, when the container of the PCR detection reagent does not include a needle tube 7, a sample injection needle tube should be provided in the PCR pre-treatment system. Optionally, the needle tube can be configured with a matching suction push rod.

[0073] The present application also proposes a PCR pre-treatment system, including a container of the PCR detection reagent.

[0074] Specifically, the container of the PCR detection reagent is described above and will not be repeated here.

[0075] Specifically, the PCR pre-treatment system further includes: a magnetic bar, which is matched with the magnetic bar sleeve 3. The magnetic bar can be made of magnetic material, or use an electromagnetic structure, etc., so that it has magnetic attraction. After the magnetic bar is placed in the magnetic bar sleeve 3, the magnetic beads contained in the groove 105 can be adsorbed on the magnetic bar sleeve 3, which is convenient for moving the magnetic beads. Preferably, through the arrangement of the magnetic poles, when the magnetic bar is erected, the magnetic poles can be respectively located on the left and right sides or the front and back sides of the magnetic bar. At this time, when the magnetic bar is placed in the magnetic bar sleeve and the magnetic beads are adsorbed, the magnetic beads are easily attached to the surrounding side of the magnetic bar sleeve rather than the bottom, which can effectively adsorb the magnetic beads over a larger area and is not affected by the sharp structural design of the end of the magnetic bar sleeve.

[0076] Optionally, the PCR pre-treatment system further includes: a first suction push rod and a second suction push rod. The first suction push rod has a connection portion that is convenient for connecting and disconnecting with the first piston 401 of the pipette 4; the second suction push rod has a connection portion that is convenient for connecting and disconnecting with the second piston 703 of the needle tube 7. The suction push rod controls the operation of the first piston 401 and the second piston 703 to complete the absorption or release of the reagent. Preferably, since the needle tube 7 and the pipette 4 usually do not work at the same time, the first suction push rod and the second suction push rod can be set as the same component, which can be connected to the first piston 401 and the second piston 703 respectively. Preferably, since the needle tube 7, the magnetic rod and the pipette 4 usually do not work at the same time, a connecting portion that cooperates with the pipette 4 and the needle tube 7 can be provided on the magnetic rod, and the magnetic rod is used as a suction push rod to be connected to the first piston 401 and the second piston 703 respectively. That is, the magnetic rod has multiple functions and can work with the magnetic rod sleeve 3 as well as the pipette 4 and the needle tube 7, thereby reducing equipment requirements and costs and simplifying the equipment.

[0077] Optionally, the connection portion can be connected and disconnected with the first plug-in portion 402 of the first piston 401 and the second plug-in portion 704 of the second piston 703 by means of an inverted connection. The first plug-in portion 402 and the second plug-in portion 704 are inner cavities with a smaller neck and a larger bottom, and the connection portions of the first suction push rod and the second suction push rod are protruding structures whose shapes match the first plug-in portion 402 and the second plug-in portion 704, respectively. When connection is required, the first suction push rod and the second suction push rod move downward and insert the connecting parts into the first plug-in part 402 and the second plug-in part 704; when disconnection is required, the first piston 401 and the second piston 703 move upward and come into contact with the piston stop bulge set at the top of the tube cavity of the pipette 4 and the needle body 701 and are blocked. When the first suction push rod and the second suction push rod continue to move upward, the connecting parts of the first suction push rod and the second suction push rod will be easily disengaged from the first plug-in part 402 and the second plug-in part 704, and the first suction push rod and the second suction push rod are disconnected from the first piston 401 and the second piston 703.

[0078] In this embodiment, the PCR pre-treatment system uses a first suction push rod and a second suction push rod to cooperate with the pipette 4 and the needle 7 respectively to complete the suction or release work of the reagent.

[0079] Optionally, the PCR pre-treatment system further includes: at least two mechanical arms, the mechanical arms are used to grab the magnetic rod sleeve 3, the pipette 4, the magnetic rod and / or the suction push rod. In this embodiment, two mechanical arms are used, one of which is used to grab, move, and control the magnetic rod, the first suction push rod, and the second suction push rod, and the other is used to grab, move, and control the magnetic rod sleeve, the pipette, and the needle. Those skilled in the art can design the number of mechanical arms according to actual needs, such as: when the magnetic rod, the first suction push rod, and the second suction push rod are set as the same component, the number of mechanical arms can be simplified to two, one for grabbing, moving, and controlling the component, and the other for grabbing, moving, and controlling the magnetic rod sleeve, the pipette, and the needle; or, according to the processing flow of the pre-treatment, the same mechanical arm can be used to control multiple components one by one, and the number of mechanical arms can be simplified to two. In this embodiment, the mechanical arm controls the magnetic rod and the suction push rod by grabbing; the mechanical arm can also be set as an integrated control device with the magnetic rod or the suction push rod.

[0080] In this embodiment, the robot arm controls the needle tube 7, the magnetic rod, the magnetic rod sleeve 3 and the pipette 4 to move horizontally and / or vertically, including:

[0081] Vertical stroke (Z-axis direction movement stroke): one robot arm controls the magnetic rod sleeve, and the other robot arm controls the magnetic rod, so that the magnetic rod and the magnetic rod sleeve can complete the following actions: 1) synchronously insert and withdraw from the magnetic rod sleeve receiving groove; 2) insert and withdraw the magnetic rod from the magnetic rod sleeve 3; 3) control the magnetic rod sleeve 3 to insert and withdraw from its receiving groove; 4) the magnetic rod sleeve 3 shakes up and down in the groove to stir. One robot arm fixes the pipette 4, and the other robot arm controls the first suction push rod to complete the following actions: 1) the first suction push rod is connected or disconnected with the first plug-in portion 402 of the first piston 401; 2) the first piston 401 is controlled to move up or down in the pipette 4; 3) the pipette 4 is withdrawn and inserted from its receiving groove. One mechanical arm fixes the needle tube, and the other mechanical arm controls the second suction push rod to complete the following actions: 1) the second suction push rod is connected or disconnected with the second plug-in portion 704 of the second piston 703; 2) the second piston 703 is controlled to move up or down in the needle tube body 701; 3) the needle tube 7 is withdrawn from or inserted into the needle tube receiving groove 6. The magnetic rod sleeve 3 and the pipette 4 can be withdrawn from or inserted into the corresponding receiving groove separately or together.

[0082] Horizontal stroke (movement stroke in the X-axis and Y-axis directions): one robotic arm controls the magnetic rod, and the other robotic arm controls the magnetic rod sleeve 3, so that the magnetic rod and the magnetic rod sleeve 3 can be synchronously moved horizontally in the closed space of the container to reach the specified position; when one robotic arm controls the magnetic rod alone, the magnetic rod can be horizontally moved to reach the specified position, for example: the position of the magnetic rod sleeve 3 or the magnetic rod storage position. When one robotic arm controls the magnetic rod sleeve 3 alone, the magnetic rod sleeve 3 can be horizontally moved in the closed space of the container to reach the specified position, for example: the position of a groove that needs to be stirred. When one robotic arm controls the pipette 4 and the other robotic arm controls the first suction push rod, the pipette 4 can be synchronously moved horizontally in the closed space of the container with the first suction push rod to reach the specified position. When one robotic arm controls the pipette 4 alone, the pipette 4 can be horizontally moved in the closed space of the container to reach the specified position. When one robotic arm controls the needle tube 7 and the other robotic arm controls the second suction push rod, the needle tube 7 can be moved horizontally in synchronization with the second suction push rod in the closed space of the container to reach the specified position; when one robotic arm controls a single needle tube 7, the needle tube 7 can be moved horizontally in the closed space of the container to reach the specified position.

[0083] It should be noted that the vertical stroke and horizontal stroke described above may occur separately or simultaneously, and the list is not exhaustive. Those skilled in the art may set a robotic arm to control the movement of corresponding components according to the requirements of the PCR processing steps.

[0084] In this embodiment, the pre-processing process is as follows:

[0085] The first mechanical arm grabs the needle tube 7 and moves it to the corresponding position, and the second mechanical arm controls the second suction push rod to make its connecting part enter the second plug-in part 704 of the second piston 703 to complete the inverted connection. After controlling the second piston 703 to move upward to absorb the sample to be tested, the first mechanical arm moves the needle tube 7 to the top of the piercing identification area of ​​the flexible sealing film 2, controls the needle tube 7 to pierce the flexible sealing film 2, and the telescopic needle protection cover 702 shrinks. The second mechanical arm controls the second suction push rod to move downward and push the second piston 703 to send the sample to be tested into the first groove 105a filled with lysis solution; then the needle tube 7 is pulled out, the telescopic needle protection cover 702 is reset, and the flexible sealing film 2 self-rebounds and seals.

[0086] If necessary, the first robot arm and the second robot arm can also control the needle tube 7 to absorb the corresponding reagents from the groove storing the sample processing reagent components such as proteinase K and send them into the first groove 105a.

[0087] Subsequently, the second mechanical arm drives the second suction push rod and the second piston 703 upward until they are blocked by the piston stop convex, the second suction push rod and the second piston 703 are disconnected, and the suction push rod is returned to the storage position. The first mechanical arm returns the needle tube 7 to the needle tube accommodating groove 6.

[0088] The second robotic arm grabs the magnetic rod sleeve 3 and controls the magnetic rod sleeve 3 so that its tip cuts through the thin sealing film of the groove in the reagent containing part 101 for storing the lysate, magnetic beads, washing solution and eluent required for the current pre-treatment.

[0089] Subsequently, the second robot arm controls the magnetic rod sleeve 3 to move to above the first groove 105a, and then moves downward into the first groove 105a to stir and mix for lysis.

[0090] After the lysis is completed in the first groove 105a, the first robot grabs the magnetic bar and places it into the magnetic bar sleeve 3, and the second robot grabs the magnetic bar sleeve 3, so that the magnetic bar and the magnetic bar sleeve 3 are synchronously moved to the top of the second groove 105b, and the magnetic bar and the magnetic bar sleeve 3 are placed in the second groove 105b containing the magnetic bead solution. The magnetic beads are adsorbed to the surface of the magnetic bar sleeve 3 due to the magnetic force of the magnetic bar. Since the magnetic poles are set on the left and right or front and back sides of the magnetic bar, the magnetic beads can be adsorbed on the left and right or front and back sides of the magnetic bar over a large area and move to the first groove 105a with the magnetic bar sleeve 3. The first robot controls the magnetic bar to withdraw, and after the magnetic force disappears, the magnetic beads are scattered in the first groove 105a. The second robot controls the magnetic bar sleeve 3 to stir in the first groove 105a, so that the lysed nucleic acid adheres to the surface of the magnetic beads. Subsequently, the first robotic arm controls the magnetic bar to be placed into the magnetic bar sleeve 3 again, and the magnetic beads adsorbed with nucleic acid are adsorbed onto the surface of the magnetic bar sleeve 3 again. The first robotic arm and the second robotic arm respectively control the magnetic bar and the magnetic bar sleeve 3 to be synchronously moved to the top of the third groove 105c containing the washing liquid, and the magnetic bar and the magnetic bar sleeve 3 are placed into the third groove 105c. Subsequently, the first robotic arm controls the magnetic bar to be withdrawn, and the magnetic beads adsorbed with nucleic acid are scattered into the washing liquid in the third groove 105c. The second robotic arm controls the magnetic bar sleeve 3 to stir in the third groove 105c for washing.

[0091] If necessary, wash the beads multiple times.

[0092] After washing is completed in the third groove 105c, the first robot arm controls the magnetic bar to be placed in the magnetic bar sleeve 3, and the magnetic beads adsorbed with nucleic acid are adsorbed again on the surface of the magnetic bar sleeve 3. The first robot arm and the second robot arm respectively control the magnetic bar and the magnetic bar sleeve 3 to move synchronously to the top of the fourth groove 105d filled with the eluent. The magnetic bar and the magnetic bar sleeve 3 are placed in the fourth groove 105d, and then the first robot arm controls the magnetic bar to withdraw, and the magnetic beads adsorbed with nucleic acid are scattered into the eluent in the fourth groove 105d. The second robot arm controls the magnetic bar sleeve 3 to stir in the fourth groove 105d for elution.

[0093] After elution is completed in the fourth groove 105d, the nucleic acid and the magnetic beads are separated. The first robotic arm controls the magnetic bar to be placed in the magnetic bar sleeve, and the magnetic beads without nucleic acid adsorbed are adsorbed to the surface of the magnetic bar sleeve 3 again. The first robotic arm and the second robotic arm respectively control the magnetic bar and the magnetic bar sleeve 3 to move synchronously away from the fourth groove 105d. At this time, the nucleic acid sample solution to be tested is obtained in the fourth groove 105d. The first robotic arm and the second robotic arm synchronously place the magnetic bar and the magnetic bar sleeve 3 back to the magnetic bar sleeve accommodating groove 102, the first robotic arm controls the magnetic bar to withdraw from the magnetic bar sleeve 3, and the magnetic beads are scattered in the magnetic bar sleeve accommodating groove 102. The first robotic arm places the magnetic bar back to the magnetic bar storage position in the system. Since the magnetic bar does not directly contact the sample or solution, it can be used for the next detection at any time.

[0094] The second robot arm grabs the pipette 4 and controls the pipette 4 so that its tip cuts through the thin sealing films of the test tubes needed for this test, the grooves for storing paraffin oil, and the pre-amplification grooves (if necessary) in the amplification tube accommodating portion 104 .

[0095] The first robotic arm grabs the first suction push rod and moves it to the position of the pipette 4, and then moves down to connect with the first plug-in portion 402 of the first piston 401. The second robotic arm controls the pipette 4 to exit the pipette accommodating groove 103 and move to the top of the fourth groove 105d, and then moves down into the fourth groove 105d. The first robotic arm controls the first suction push rod to absorb the nucleic acid sample solution in the fourth groove 105d. Subsequently, the second robotic arm controls the pipette 4 to move up and leave the fourth groove 105.

[0096] If necessary, pre-amplification can be performed. The second mechanical arm controls the pipette 4 to move to the top of the fifth groove 105e where the pre-amplification reagent is stored, and the pipette 4 moves downward into the fifth groove 105e. The first mechanical arm controls the first suction push rod to push the nucleic acid sample solution into the fifth groove 105e. The first and second mechanical arms respectively control the pipette 4 and the first suction push rod to absorb a little paraffin oil from the sixth groove 105f where the paraffin oil is stored to the fifth groove 105e for pre-amplification. The paraffin oil can prevent the solution from evaporating.

[0097] After the pre-amplification is completed, the second robot arm controls the pipette 4 to move to the top of the fifth groove 105e, and then moves downward into the fifth groove 105e. The first robot arm controls the first suction push rod to absorb the sample solution that has completed the pre-amplification. Subsequently, the first robot arm controls the pipette 4 to move upward from the fifth groove 105e and move to the top of the test tube 106 where the PCR reagent is stored, and then moves downward into the test tube 106. The first robot arm controls the first suction push rod to push the sample solution that has completed the pre-amplification into the test tube 106. The first and second robot arms respectively control the pipette 4 and the first suction push rod to absorb a little paraffin oil from the sixth groove 105f where the paraffin oil is stored into the test tube 106 to perform an amplification reaction. The paraffin oil can prevent the solution from evaporating.

[0098] If pre-amplification is not required, the pre-amplification step is skipped, and the pipette 4 directly transfers the nucleic acid sample solution from the fourth groove 105 d and places it into the test tube 106 containing the PCR reagent.

[0099] Optionally, the pipette 4 may also move a little paraffin oil into the fourth groove 105d where the nucleic acid sample solution may remain, so as to achieve a further sealing effect and prevent the nucleic acid from escaping.

[0100] In the description of this application, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood according to specific circumstances.

[0101] In the present application, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

[0102] In the description of this embodiment, the terms "upper", "lower", "left", "right", etc., and the orientation or position relationship are based on the orientation or position relationship shown in the drawings, and are only for the convenience of description and simplification of operation, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as a limitation of the present application. In addition, the terms "first" and "second" are only used to distinguish in the description and have no special meaning.

[0103] The above embodiments are only used to illustrate the technical solution of the present application but not to limit it. The present application is described in detail with reference to the preferred embodiments. It should be understood by those skilled in the art that the technical solution of the present application can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present application, and should be included in the scope of the claims of the present application.

Claims

1. A container for PCR detection reagents, characterized in that: include: A box body, a flexible sealing film, a magnetic rod sleeve, a pipette and a thin sealing film; the box body includes a reagent accommodating part and an amplification tube accommodating part, the reagent accommodating part is provided with a plurality of grooves, the amplification tube accommodating part is provided with at least one test tube, the thin sealing film is arranged at the openings of the grooves and the test tubes, so that the grooves or the test tubes are individually sealed; the flexible sealing film is fixedly and sealedly connected to the peripheries of the magnetic rod sleeve and the pipette, the flexible sealing film covers the top surface of the box body, and together with the box body, the magnetic rod sleeve and the pipette, forms a closed space.

2. The container of PCR detection reagent according to claim 1, characterized in that: Also includes: A cover frame is fixedly and sealingly connected to the flexible sealing film and the box body.

3. The container of PCR detection reagent according to claim 2, characterized in that: The flexible sealing film is an airtight, watertight, elastic film with good elongation at break performance; in the stretched state, the elongation at break performance of the flexible sealing film should satisfy the requirement that the magnetic rod sleeve and the pipette are withdrawn from their respective accommodating grooves separately or together to complete a vertical stroke, and / or a horizontal stroke without breaking; after being punctured by a needle, the flexible sealing film can rebound by itself and reseal the needle hole.

4. The container of PCR detection reagent according to claim 3, characterized in that: The flexible sealing film is made of TPV, TPE, TPR, silicone or latex.

5. The container of PCR detection reagent according to claim 3, characterized in that: The edge of the flexible sealing film in contact with the side wall of the box body is provided with at least one piercing identification area.

6. The container of PCR detection reagent according to claim 3, characterized in that: The box body or the flexible sealing film is provided with an identification code.

7. The container of PCR detection reagent according to claim 2, characterized in that: The cover frame, the flexible sealing film, the magnetic rod sleeve and the pipette are integrally formed.

8. The container of PCR detection reagent according to claim 2, characterized in that: The magnetic rod sleeve It is a semi-enclosed tubular body with only the top end open; the pipette is a cavity structure with the top and bottom ends open, a first piston is arranged in the cavity, and the first piston seals the liquid containing area of ​​the pipette from the top.

9. The container of PCR detection reagent according to claim 8, characterized in that: The bottom end of the magnetic rod sleeve and / or the pipette has a pointed end.

10. The container of PCR detection reagent according to claim 8, characterized in that: An annular convex structure is circumferentially arranged at the middle part of the outer wall of the magnetic rod sleeve, and the annular convex structure circumferentially protrudes from the outer wall of the magnetic rod sleeve below it.

11. The container of PCR detection reagent according to claim 1, characterized in that: The box body further comprises: a magnetic rod sleeve accommodating groove and a pipette accommodating groove. The magnetic rod sleeve accommodating groove is used to place the magnetic rod sleeve, and the pipette accommodating groove is used to place the pipette.

12. The container of PCR detection reagent according to claim 11, characterized in that: The reagent containing part and the amplification tube containing part are respectively located at two sides of the box body; the magnetic rod sleeve containing groove is arranged in the reagent containing part.

13. The container of PCR detection reagent according to claim 11, characterized in that: The reagent containing part and the amplification tube containing part are an integral structure with the wall of the box body; or, although they are separate structures, they are fixedly sealed and connected by means of buckling or the like.

14. The container of PCR detection reagent according to claim 1, characterized in that: The thin sealing film can be one or more sheets, and is made of aluminum-plastic material.

15. The container of PCR detection reagent according to any one of claims 1 to 14, characterized in that: Also includes: The needle tube comprises a needle tube body, a steel needle and a second piston.

16. The container of PCR detection reagent according to claim 15, characterized in that: The needle tube also includes: a telescopic needle protection sleeve, the steel needle is wrapped by the telescopic needle protection sleeve, one end of which is fixed to the needle tube body and the other end is a free end.

17. The container of PCR detection reagent according to claim 15, characterized in that: It also includes: a needle tube accommodating groove; the needle tube accommodating groove is arranged beside the box body outside the closed space of the container, and is used to place the needle tube.

18. The container of PCR detection reagent according to claim 15, characterized in that: Paraffin oil is placed in any one of the grooves.

19. A PCR pre-treatment system, characterized in that: A container comprising a PCR detection reagent as described in any one of claims 1-18, further comprising: a magnetic rod, a first suction push rod, a second suction push rod, and at least two mechanical arms, wherein the magnetic rod has magnetic attraction and can be placed in a magnetic rod sleeve; the first suction push rod can be connected and disconnected with the first piston of the pipette, and the second suction push rod can be connected and disconnected with the second piston of the syringe; the mechanical arm can be used to control the movement of the magnetic rod sleeve, the pipette, the syringe, the magnetic rod and / or the suction push rod.

20. The PCR pre-treatment system according to claim 19, characterized in that: The first suction push rod and the second suction push rod are the same component, and the component can be connected to the first piston and the second piston respectively.

21. The PCR pre-treatment system according to claim 19, characterized in that: The magnetic rod and the first suction push rod are the same component, which has magnetic attraction, can be placed in the magnetic rod sleeve and can be connected to the first piston.

22. The PCR pre-treatment system according to claim 19, characterized in that: The magnetic rod, the first suction push rod and the second suction push rod are the same component, which has magnetic attraction, can be placed in the magnetic rod sleeve and can be connected to the first piston and the second piston respectively.

23. The PCR pre-treatment system according to any one of claims 19 to 22, characterized in that: When the magnetic bar is erected, the magnetic poles are respectively located on the left and right sides or the front and rear sides of the magnetic bar.

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