A pollution prevention device and method for real-time fluorescent PCR

Abstract

The present application relates to a kind of anti-pollution device and method for real-time fluorescent PCR.The device includes fixed support, first movable support, plunger pump, second movable support, motor, movement module, sleeve, suction head, first test tube, second test tube, third test tube and fourth test tube.The second movable support is installed on the fixed support and is slidably connected with the second sliding support along the Z-axis direction.The sleeve is vertically arranged and detachably installed on the second movable support.One end of the sleeve is connected with the plunger pump through a pipeline, and the other end is detachably connected with the suction head.The first movable support is installed on the fixed support through the movement module and is located below the second movable support.The first test tube, second test tube, third test tube and fourth test tube are placed on the first movable support.Three seals are achieved, which effectively prevents pollution and is fully automatic with high efficiency.The cooperation of the cover and the suction head dismounting plate improves the safety performance while preventing pollution.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of PCR technology, and more particularly, to a pollution prevention device and method for real-time fluorescent PCR. BACKGROUND

[0002] Polymerase chain reaction (PCR) technology is a method for rapid amplification of specific target nucleic acid fragments in vitro, which is usually composed of three basic reaction steps of denaturation, annealing and extension, and the amplification of target nucleic acid is achieved by temperature cycling.

[0003] Real-time fluorescent PCR technology is a nucleic acid / gene detection technology developed on the basis of PCR technology, which is a method for monitoring the entire PCR process by real-time detection of fluorescence signal by adding fluorescently labeled probes or fluorescent dyes in the PCR reaction system, and finally performing qualitative or quantitative analysis of nucleic acid or gene by specific fluorescence signal / standard curve / internal standard.

[0004] The biggest feature of real-time fluorescent PCR instrument is that it has large amplification capacity and high sensitivity, and the operation is complex, so pollution often occurs during operation, even a small amount of pollution can cause false positive, and a slight mistake can cause false negative. For a long time, how to take effective measures to prevent pollution to the maximum extent is a major design requirement for real-time fluorescent PCR instrument.

[0005] Chinese patent CN111197004A discloses a kind of full-automatic real-time fluorescent quantitative PCR workstation, realizes the full-automatic operation of PCR process, but the device is easy to cause pollution during operation, causes the problems such as inaccurate measurement. SUMMARY

[0006] To overcome at least one of the above-mentioned defects in the prior art, the present application provides a pollution prevention device and method for real-time fluorescent PCR, which realizes multiple sealing of the test tube storing the sample and effectively prevents pollution of the sample.

[0007] To solve the above technical problems, the technical solution adopted by the present invention is: an anti-contamination device for real-time fluorescence PCR, comprising a fixed bracket, a first movable bracket for storing test tubes, a plunger pump, a second movable bracket for fixing a sleeve and a pipette tip, a motor for driving the second movable bracket to move, a motion module for driving the first movable bracket to move in a horizontal plane, a sleeve, a pipette tip, a first test tube for storing samples, a second test tube for storing a suction ball used for a ball seal, a third test tube for storing a liquid used for a liquid seal, and a fourth test tube for storing a pipette tip, the second movable bracket is mounted on the fixed bracket and is slidably connected to the second sliding bracket along the Z-axis direction, the sleeve is vertically arranged and detachably mounted on the second movable bracket, one end of the sleeve is connected to the plunger pump through a pipe, and the other end is detachably connected to the pipette tip; the first movable bracket is mounted on the fixed bracket through the motion module and is located below the second movable bracket; the first test tube, the second test tube, the third test tube, and the fourth test tube are all vertically arranged and detachably mounted on the first movable bracket. In the present invention, the first movable bracket is driven by the motion module to move left and right or forward and backward on the horizontal plane, so as to move the first test tube, the second test tube, the third test tube, and the fourth test tube placed on the first movable bracket to the bottom of the sleeve, so as to realize the actions of the suction head sucking liquid and the sleeve sucking the suction ball, thereby moving the liquid and the suction ball into the first test tube; the second movable bracket is located directly above the first movable bracket, and the second movable bracket is driven by the motor to move up and down along the Z-axis direction, so as to realize the suction from the test tube or dripping liquid into the test tube when descending; the anti-pollution device provided by the present invention, when in use, the suction head is placed in the fourth test tube, and the first movable bracket is moved to make the fourth test tube suck liquid. Located just below the sleeve, the motor drives the second movable bracket to descend, so that one end of the bottom of the sleeve presses down the suction head to pass through the hole of the suction head, thereby realizing that the suction head is sleeved on the sleeve; thereafter, the first movable bracket and the second movable bracket are repeatedly moved to move the oil stored in the third test tube into the first test tube, realizing the first sealing of the sample; then, the suction head is detached from the fourth test tube, and the suction ball stored in the second test tube is moved into the first test tube through the sleeve, realizing the second sealing of the sample. In addition, the outer cover of the first test tube can be sealed as needed to realize the third sealing; multiple sealings effectively prevent the contamination of the sample, and the fully automatic operation has high work efficiency.

[0008] In one embodiment, the second movable bracket includes a long sleeve holder disposed transversely and slidably connected to the fixed bracket along the Z-axis, and a long tip removal plate disposed transversely and slidably connected to the fixed bracket along the Z-axis and located directly below the sleeve holder. A plurality of first mounting holes are provided at intervals along the length of the sleeve holder, and a plurality of second mounting holes are provided at intervals along the length of the tip removal plate, corresponding one-to-one to the positions of the first mounting holes. One end of the sleeve is fixed in the first mounting hole, and the other end is passed through the second mounting hole and connected to the tip. The sleeve holder is used to secure the sleeve, and the tip removal plate is used to push the tip off when removing the tip. The tip removal plate can also be used to press the cap to ensure that the cap fits tightly against the first test tube.

[0009] In one embodiment, a first annular boss is provided in the second mounting hole, and a second annular boss is provided at the end of the nozzle connected to the sleeve, configured to abut against the first annular boss. When the nozzle needs to be removed, the second motor drives the nozzle removal plate downward, causing the first annular boss to abut against the second annular boss on the nozzle, thereby pushing the nozzle off and achieving automatic removal.

[0010] In one embodiment, the depth values ​​of the first annular bosses in the plurality of second mounting holes are divided into at least two groups. Dividing the depths of the first annular bosses in the plurality of through holes into at least two depth values ​​allows for batch removal of the nozzle tips, which is more labor-saving and safer.

[0011] In one embodiment, the motor includes a first motor and a second motor; the output shaft of the first motor is connected to the sleeve frame for driving the sleeve frame to move up and down along the Z-axis; the output shaft of the second motor is connected to the suction head detachment plate for driving the suction head detachment plate to move up and down along the Z-axis.

[0012] In one embodiment, the device further includes a cap for covering the opening of the first test tube, the cap being detachably connected to the first test tube. After the liquid seal and ball seal are complete, the cap is replaced to provide a third level of sealing. The cap is held in place by a tip release plate, preventing it from popping during heating, effectively preventing contamination and enhancing safety.

[0013] In one embodiment, a sealing ring is provided at the orifice of the first test tube; and a ball seal retainer is provided within the first test tube near one end of the orifice for retaining a suction ball. The sealing ring is used to achieve a sealed connection between the cover and the orifice of the test tube; and the ball seal retainer is used to retain the suction ball.

[0014] In one of the embodiments, the first movable support is provided with a plurality of placement slots at intervals, the plurality of placement slots are arranged in multiple rows and are parallel to each other, the interval distance of the plurality of placement slots in each row corresponds to the interval distance of the plurality of first mounting through holes one by one; the first test tube, the second test tube, the third test tube and the fourth test tube are respectively placed in the placement slots. The placement slots are arranged in multiple rows, the test tubes of the same function are placed in the same row, which facilitates simultaneous operation and improves efficiency; the interval distance between each row of placement slots corresponds to the interval distance between the sleeves one by one, so that the plurality of sleeves or suction heads can be operated simultaneously, improving the work efficiency.

[0015] In one of the embodiments, the motion module comprises an X-axis movement system and a Y-axis movement system, and the first movable support is connected with the X-axis movement system and the Y-axis movement system; each row of placement slots of the first movable support is arranged along the X-axis direction, and the sleeve rack and the suction head dismounting plate are also arranged along the X-axis direction.

[0016] In one of the embodiments, a third motor is further included, the bottom of the second test tube is provided with a through hole, a thimble is arranged in the through hole, one end of the thimble is used for pushing the suction ball, and the other end is connected with the output shaft of the third motor. The suction ball is pushed out by the thimble, and the sleeve can suck the suction ball.

[0017] The application further provides a pollution prevention method for real-time fluorescent PCR, which uses the pollution prevention device described above and comprises the following steps:

[0018] S1. Oil sealing: the motion module is started to move the first movable support to below the second movable support, and the fourth test tube provided with the suction head is just located below the sleeve; the first motor and the second motor are started to move the second support and the sleeve downward at the same time until the sleeve is connected with the suction head, and the suction head is sleeved on the sleeve; then, the first motor and the second motor are started to move the second support upward, the motion module is started to move the third test tube provided with oil to below the suction head; the first motor and the second motor are started again, the suction head moves downward into the third test tube, the plunger pump is started to make the suction head suck a small amount of oil; finally, the first motor and the second motor are started, the suction head moves upward, the motion module is started to move the first test tube provided with a sample to below the suction head; the first motor and the second motor are started again, the suction head moves downward, the plunger pump is depressurized, the oil in the suction head drops into the first test tube, and the first layer sealing is completed;

[0019] S2. Ball sealing: First, the motion module is activated to move the fourth test tube to the position directly below the suction head. The second motor is activated to move the suction head removal plate downward. The first annular boss on the suction head removal plate abuts against the second annular boss on the suction head, and the suction head is ejected and falls into the fourth test tube. The motion module is activated to move the second test tube containing the sealing ball to the position directly below the sleeve. Then, the first and second motors are activated to move the second movable bracket downward, so that the suction nozzle of the sleeve contacts the sealing ball. The plunger pump is activated to attract the sealing ball. The first and second motors are activated to move the second movable bracket upward as a whole. The motion module is activated to move the first test tube to the position directly below the sleeve. Finally, the first and second motors are activated to move the sleeve downward, the plunger pump is depressurized, and the sealing ball falls into the first test tube, completing the second layer of sealing.

[0020] S3. Capping: Place the cap on the first test tube, start the second motor, and move the tip removal plate downward so that the tip removal plate presses against the cap to ensure close contact between the removal plate and the first test tube; completing the third layer of sealing.

[0021] Compared with the prior art, the present invention has the following beneficial effects: the anti-pollution device for real-time fluorescence PCR provided by the present invention can realize triple sealing of oil seal, ball seal and cover seal, effectively preventing the PCR tube from being contaminated, and the triple sealing is fully automatic and efficient; by providing multiple sleeves, pipette tips and corresponding test tubes, multiple samples can be operated sequentially with high efficiency; through the cooperation of the cover and the pipette tip detachment plate, while preventing contamination, the safety performance is also improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0023] Figure 2 It is a schematic structural diagram of the sleeve and the second movable bracket of the present invention.

[0024] Figure 3 It is a schematic structural diagram of the first movable bracket of the present invention.

[0025] Figure 4 It is a schematic diagram of the first test tube structure of the present invention.

[0026] Figure 5 It is a schematic diagram of the connection between the suction head and the sleeve of the present invention.

[0027] Description of the drawings: 1. Fixed bracket; 2. First movable bracket; 21. Placement slot; 3. Plunger pump; 4. Second movable bracket; 41. Sleeve rack; 42. Tip detachable plate; 5. Motor; 51. First motor; 52. Second motor; 6. Motion module; 7. Sleeve; 8. Tip; 81. Second annular boss; 9. First test tube; 91. Ball seal position. DETAILED DESCRIPTION

[0028] The accompanying drawings are for illustrative purposes only and are not to be construed as limiting the present invention. To better illustrate the present embodiment, some components in the accompanying drawings may be omitted, enlarged, or reduced in size, and do not represent actual product dimensions. Those skilled in the art will appreciate that some well-known structures and their descriptions may be omitted from the accompanying drawings. The positional relationships depicted in the accompanying drawings are for illustrative purposes only and are not to be construed as limiting the present invention.

[0029] like Figures 1 to 3 As shown, an anti-contamination device for real-time fluorescence PCR includes a fixed support 1, a first movable support 2 for storing test tubes, a plunger pump 3, a second movable support 4 for fixing a sleeve 7 and a pipette tip 8, a motor 5 for driving the second movable support 4 to move, a motion module 6 for driving the first movable support 2 to move in a horizontal plane, a sleeve 7, a pipette tip 8, a first test tube 9 for storing a sample, a second test tube for storing a suction ball used for a ball seal, a third test tube for storing a liquid used for a liquid seal, and a fourth test tube for storing the pipette tip 8. The second movable support 4 is mounted on the fixed support 1 and is slidably connected to the second sliding support along the Z-axis direction. The sleeve 7 is vertically arranged and detachably mounted on the second movable support 4. One end of the sleeve 7 is connected to the plunger pump 3 through a pipe, and the other end is detachably connected to the pipette tip 8. The first movable support 2 is mounted on the fixed support 1 through the motion module 6 and is located below the second movable support 4. The first test tube 9, the second test tube, the third test tube, and the fourth test tube are all vertically arranged and detachably mounted on the first movable support 2. In the present invention, the first movable bracket 2 is driven by the motion module 6 to move left and right or up and down on the horizontal plane, so as to move the first test tube 9, the second test tube, the third test tube, and the fourth test tube placed on the first movable bracket 2 to the bottom of the sleeve 7, so as to realize the actions of the suction head 8 sucking liquid and the sleeve 7 sucking the ball, thereby moving the liquid and the suction ball into the first test tube 9; the second movable bracket 4 is located directly above the first movable bracket 2, and the second movable bracket 4 is driven by the motor 5 to move up and down along the Z-axis direction to realize sucking liquid from the test tube or dripping liquid into the test tube when descending; the anti-pollution device provided by the present invention, when in use, the suction head 8 is placed in the fourth test tube, and the first movable bracket 2 is moved so that the fourth test tube is located Directly below the sleeve 7, the motor 5 drives the second movable bracket 4 to descend, so that one end of the bottom of the sleeve 7 presses down the suction head 8 to pass through the hole of the suction head 8, thereby realizing that the suction head 8 is sleeved on the sleeve 7; thereafter, the first movable bracket 2 and the second movable bracket 4 are repeatedly moved to move the oil stored in the third test tube to the first test tube 9, realizing the first sealing of the sample; then, the suction head 8 is removed to the fourth test tube, and the suction ball stored in the second test tube is moved to the first test tube 9 through the sleeve 7, realizing the second sealing of the sample. In addition, as needed, the outer cover of the first test tube 9 can be sealed to realize the third sealing; multiple sealings effectively prevent the contamination of the sample, and the fully automatic operation has high work efficiency.

[0030] In some embodiments, as Figure 2 As shown, the second movable bracket 4 includes a long sleeve holder 41 disposed transversely and slidably connected to the fixed bracket 1 along the Z-axis, and a long tip removal plate 42 disposed transversely and slidably connected to the fixed bracket 1 along the Z-axis and located directly below the sleeve holder 41. A plurality of first mounting holes are spaced apart along the length of the sleeve holder 41, and a plurality of second mounting holes are spaced apart along the length of the tip removal plate 42, corresponding one-to-one to the positions of the first mounting holes. One end of the sleeve 7 is fixed in the first mounting hole, and the other end is inserted into the second mounting hole and connected to the tip 8. The sleeve holder 41 is used to fix the sleeve 7, and the tip removal plate 42 is used to push the tip 8 off when removing it. The tip removal plate 42 can also be used to press the cover to ensure that the cover fits tightly against the first test tube 9.

[0031] In some embodiments, a slide groove is provided on the fixed bracket 1, and a slider is provided on the sleeve rack 41 and the suction head removal plate 42. The slider is slidably connected to the slide groove and can also achieve a limiting effect to prevent the motor 5 from shaking when driving the sleeve rack 41 and the suction head removal plate 42 to move up and down, thereby improving stability during movement.

[0032] In one embodiment, Figure 5 As shown, a first annular boss is provided in the second mounting hole, and a second annular boss 81 is provided at the end of the nozzle 8 connected to the sleeve 7, which is configured to abut against the first annular boss. When the nozzle 8 needs to be removed, the second motor 52 drives the nozzle removal plate 42 downward, causing the first annular boss to abut against the second annular boss 81 on the nozzle 8, thereby pushing the nozzle 8 off and achieving automatic removal of the nozzle 8.

[0033] In another embodiment, the depth values ​​of the first annular bosses in the plurality of second mounting holes are divided into at least two groups. Dividing the depths of the first annular bosses in the plurality of through holes into at least two depth values ​​allows for batch removal of the suction tips 8, which is more labor-saving and safer.

[0034] In one of the embodiments, the sleeve frame 41 is provided with 16 first mounting holes, the suction head dismounting plate 42 is provided with 16 second mounting holes, 16 sleeves 7 are mounted on the sleeve frame 41, and each sleeve 7 is provided with a corresponding suction head 8; similarly, the first movable support 2 is provided with 16 placing slots 21 in each row, and at least four rows of placing slots 21 are provided, which correspond to the first test tube 9, the second test tube, the third test tube and the fourth test tube respectively; in this way, 16 groups of samples can be operated at a time during operation, and the efficiency is high; in addition, the first annular boss is provided with three groups of depth values, and the first annular boss of each group is provided with the same depth value in at least five second mounting holes; in this way, the suction heads 8 can be dismounted in three groups, and five suction heads 8 are dismounted at a time, which is more labor-saving and safe than dismounting 16 suction heads at a time.

[0035] In one of the embodiments, as shown in Figure 1 and Figure 2 , the motor 5 includes a first motor 51 and a second motor 52; the output shaft of the first motor 51 is connected with the sleeve frame 41, and is used to drive the sleeve frame 41 to move up and down along the Z-axis direction; the output shaft of the second motor 52 is connected with the suction head dismounting plate 42, and is used to drive the suction head dismounting plate 42 to move up and down along the Z-axis direction.

[0036] In some embodiments, a cover is further included for covering the tube opening of the first test tube 9, and the cover is detachably connected with the first test tube 9. After the liquid seal and the ball seal are completed, the cover is covered, the third resealing is realized, the cover is pressed tightly by the suction head dismounting plate 42, and the cover explosion during the heating process can be prevented, which effectively prevents pollution and improves safety performance.

[0037] In one of the embodiments, a sealing ring is arranged at the tube opening of the first test tube 9; and a ball seal clamping position 91 is arranged at one end of the first test tube 9 close to the tube opening. The sealing ring is used to realize the sealing connection between the cover and the tube opening; and the ball seal clamping position 91 is used to clamp the suction ball.

[0038] As shown in Figure 4 , the first test tube 9 can be arranged in the form of a row, such as 6-tube, so as to facilitate movement and operation.

[0039] In some embodiments, as shown in Figure 1 and Figure 3As shown, the first movable bracket 2 is provided with a plurality of placement slots 21 spaced apart. The placement slots 21 are arranged in multiple rows and parallel to each other. The spacing between the placement slots 21 in each row corresponds one-to-one to the spacing between the plurality of first mounting holes. The first test tube 9, the second test tube, the third test tube, and the fourth test tube are placed in the placement slots 21, respectively. The placement slots 21 are arranged in multiple rows, with test tubes with the same function placed in the same row, facilitating simultaneous operation and improving efficiency. The spacing between the placement slots 21 in each row corresponds one-to-one to the spacing between the sleeves 7. This allows for simultaneous operation of multiple sleeves 7 or pipette tips 8, improving work efficiency.

[0040] In one embodiment, Figure 3 As shown, the motion module 6 includes an X-axis moving system and a Y-axis moving system, and the first movable bracket 2 is connected to the X-axis moving system and the Y-axis moving system respectively; each row of placement slots 21 of the first movable bracket 2 is arranged along the X-axis direction, and the sleeve rack 41 and the suction head removal plate 42 are also arranged along the X-axis direction.

[0041] In another embodiment, a third motor is further included. The bottom of the second test tube is provided with a through hole, through which a pin is inserted. One end of the pin is used to support the suction ball, and the other end is connected to the output shaft of the third motor. The pin pushes the suction ball out, allowing the sleeve 7 to absorb the suction ball.

[0042] In another embodiment, a contamination prevention method for real-time fluorescent PCR is provided, using the above-mentioned contamination prevention device, comprising the following steps:

[0043] S1. Oil seal: Start the motion module 6 to move the first movable bracket 2 to the bottom of the second movable bracket 4, and make the fourth test tube with the suction head 8 just under the sleeve 7; start the first motor 51 and the second motor 52 to move the second bracket and the sleeve 7 downward at the same time until the sleeve 7 is connected with the suction head 8 and the suction head 8 is sleeved on the sleeve 7; then, start the first motor 51 and the second motor 52 to move the second bracket upward, start the motion module 6, and move the third test tube with oil to The first motor 51 and the second motor 52 are started again, the suction head 8 moves down into the third test tube, and the plunger pump 3 is started to make the suction head 8 absorb a small amount of oil. Finally, the first motor 51 and the second motor 52 are started, the suction head 8 moves up, and the motion module 6 is started to move the first test tube 9 containing the sample to the position directly below the suction head 8. The first motor 51 and the second motor 52 are started again, the suction head 8 moves down, the plunger pump 3 is depressurized, and the oil in the suction head 8 drips into the first test tube 9, completing the first layer of sealing.

[0044] S2. Ball sealing: first, start the motion module 6, move the fourth test tube to the position directly below the suction head 8, start the second motor 52, and the suction head dismounting plate 42 moves downward, the first annular boss on the suction head dismounting plate 42 abuts against the second annular boss 81 on the suction head 8, the suction head 8 is pushed out and falls into the fourth test tube; start the motion module 6, move the second test tube containing the sealing ball to the position directly below the sleeve 7; then, start the first motor 51 and the second motor 52, and the second movable support 4 moves downward, so that the suction nozzle of the sleeve 7 contacts the sealing ball, start the plunger pump 3, and the sleeve 7 sucks the sealing ball; start the first motor 51 and the second motor 52, and the second movable support 4 moves upward as a whole; start the motion module 6, and move the first test tube 9 to the position directly below the sleeve 7; finally, start the first motor 51 and the second motor 52, and the sleeve 7 moves downward, the plunger pump 3 is depressurized, and the sealing ball falls into the first test tube 9, thus completing the second layer of sealing.

[0045] S3. Cap sealing: cover the cap on the first test tube 9, start the second motor 52, and the suction head dismounting plate 42 moves downward, so that the suction head dismounting plate 42 presses the cap, to ensure that the suction head dismounting plate is in close contact with the first test tube 9; and the third layer of sealing is completed.

[0046] In the description of the present application, it should be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the present application.

[0047] In addition, the terms "first" and "second" are only for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the technical features indicated. Therefore, the features defined as "first" and "second" can explicitly or implicitly include at least one of the features. In the description of the present application, the meaning of "a plurality of" is at least two, for example, two, three, etc., unless otherwise specifically limited.

[0048] In the present invention, unless otherwise specified or limited, the terms "installed," "connected," "connect," "fixed," etc. should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integration; mechanical connection, electrical connection; direct connection, or indirect connection through an intermediate medium; internal communication between two components, or interaction between two components, unless otherwise specified. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0049] In the present invention, unless otherwise expressly specified or limited, when a first feature is "above" or "below" a second feature, it may mean that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediary. Furthermore, when a first feature is "above," "above," or "above" a second feature, it may mean that the first feature is directly above or diagonally above the second feature, or simply means that the first feature is at a higher level than the second feature. When a first feature is "below," "below," or "below" a second feature, it may mean that the first feature is directly below or diagonally below the second feature, or simply means that the first feature is at a lower level than the second feature.

[0050] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0051] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

[0052] Obviously, the above embodiments of the present invention are merely examples for the purpose of clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Those skilled in the art will appreciate that other variations or modifications can be made based on the above description. It is not necessary and impossible to enumerate all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention are intended to be included within the scope of protection of the claims of the present invention.

Claims

1. An anti-pollution device for real-time fluorescence PCR, characterized in that: The invention comprises a fixed support (1), a first movable support (2) for storing a test tube, a plunger pump (3), a second movable support (4) for fixing a sleeve (7) and a suction head (8), a motor (5) for driving the second movable support (4) to move, a motion module (6) for driving the first movable support (2) to move on a horizontal plane, a sleeve (7), a suction head (8), a first test tube (9) for storing a sample, a second test tube for storing a suction ball used for a ball seal, a third test tube for storing a liquid used for a liquid seal, and a fourth test tube for storing a suction head (8), wherein the second The movable bracket (4) is mounted on the fixed bracket (1) and is slidably connected to the fixed bracket (1) along the Z-axis direction. The sleeve (7) is vertically arranged and detachably mounted on the second movable bracket (4). One end of the sleeve (7) is connected to the plunger pump (3) through a pipeline, and the other end is detachably connected to the suction head (8). The first movable bracket (2) is mounted on the fixed bracket (1) through a motion module (6). The first test tube (9), the second test tube, the third test tube, and the fourth test tube are all vertically arranged and detachably mounted on the first movable bracket (2). The second movable bracket (4) comprises a strip-shaped sleeve frame (41) which is arranged horizontally and connected to the fixed bracket (1) in a sliding manner along the Z-axis direction, and a strip-shaped suction head detachment plate (42) which is arranged horizontally and connected to the fixed bracket (1) in a sliding manner along the Z-axis direction. A plurality of first mounting holes are provided at intervals along the length direction of the sleeve frame (41), and a plurality of second mounting holes corresponding to the positions of the first mounting holes are provided at intervals along the length direction of the suction head detachment plate (42); one end of the sleeve (7) is fixed in the first mounting hole, and the other end is passed through the second mounting hole and is connected to the suction head ( 8) connection; a first annular boss is provided in the second mounting through hole, and one end of the suction head (8) connected to the sleeve (7) is provided with a second annular boss (81) for abutting against the first annular boss; the motor (5) comprises a first motor (51) and a second motor (52); the output shaft of the first motor (51) is connected to the sleeve frame (41) for driving the sleeve frame (41) to move up and down along the Z-axis; the output shaft of the second motor (52) is connected to the suction head detachment plate (42) for driving the suction head detachment plate (42) to move up and down along the Z-axis; It also includes a cover for covering the tube mouth of the first test tube (9), the cover being detachably connected to the first test tube (9); a sealing ring is provided at the tube mouth of the first test tube (9); and a ball seal clamping position (91) for clamping a suction ball is provided in the first test tube (9) near one end of the tube mouth. The first movable bracket (2) is provided with a plurality of placement slots (21) at intervals, the plurality of placement slots (21) are arranged in multiple rows and are parallel to each other, and the spacing between the plurality of placement slots (21) in each row corresponds to the spacing between the plurality of first mounting through holes; the first test tube (9), the second test tube, the third test tube, and the fourth test tube are respectively placed in the placement slots (21); It also includes a third motor. The bottom of the second test tube is provided with a through hole, and a thimble is passed through the through hole. One end of the thimble is used to support the suction ball, and the other end is connected to the output shaft of the third motor.

2. The anti-pollution device for real-time fluorescent PCR according to claim 1, characterized in that: There are at least two groups of depth values ​​of the first annular bosses in the plurality of second mounting through holes.

3. The anti-pollution device for real-time fluorescent PCR according to claim 1, characterized in that: The motion module (6) includes an X-axis motion system and a Y-axis motion system, and the first movable bracket (2) is connected to the X-axis motion system and the Y-axis motion system respectively; each row of placement slots (21) of the first movable bracket (2) is arranged along the X-axis direction, and the sleeve rack (41) and the suction head removal plate (42) are also arranged along the X-axis direction.

4. A contamination prevention method for real-time fluorescence PCR, characterized in that: The anti-pollution device according to any one of claims 1 to 3 is characterized by comprising the following steps: S1. Oil seal: Start the motion module (6) to move the first movable bracket (2) to the bottom of the second movable bracket (4), and make the fourth test tube with the suction head (8) just below the sleeve (7); start the first motor (51) and the second motor (52) to move the second bracket and the sleeve (7) downward at the same time until the sleeve (7) is connected to the suction head (8) and the suction head (8) is sleeved on the sleeve (7); then, start the first motor (51) and the second motor (52) to move the second bracket upward, start the motion module (6), and move the third test tube with oil to the suction head ( 8); start the first motor (51) and the second motor (52) again, the suction head (8) moves down to the third test tube, start the plunger pump (3), and make the suction head (8) absorb a small amount of oil; finally, start the first motor (51) and the second motor (52), the suction head (8) moves up, start the motion module (6), and move the first test tube (9) containing the sample to just below the suction head (8); start the first motor (51) and the second motor (52) again, the suction head (8) moves down, the plunger pump (3) releases pressure, and the oil in the suction head (8) drips into the first test tube (9), completing the first layer of sealing; S2. Ball seal: First, start the motion module (6) to move the fourth test tube to the bottom of the suction head (8), start the second motor (52), the suction head detachment plate (42) moves downward, the first annular boss on the suction head detachment plate (42) abuts against the second annular boss (81) on the suction head (8), the suction head (8) is ejected and falls into the fourth test tube; start the motion module (6) to move the second test tube equipped with the sealing ball to the bottom of the sleeve (7); then, start the first motor (51) and the second motor (52), the second The movable bracket (4) moves downward, so that the suction nozzle of the sleeve (7) contacts the sealing ball, the plunger pump (3) is started, and the sleeve (7) sucks the sealing ball; the first motor (51) and the second motor (52) are started, so that the second movable bracket (4) moves upward as a whole; the motion module (6) is started, so that the first test tube (9) moves to the bottom of the sleeve (7); finally, the first motor (51) and the second motor (52) are started, so that the sleeve (7) moves downward, the plunger pump (3) releases pressure, and the sealing ball falls into the first test tube (9), completing the second layer of sealing; S3. Capping: Place the cap on the first test tube (9), start the second motor (52), and move the tip detachment plate (42) downward so that the tip detachment plate (42) presses against the cap to ensure that the tip detachment plate (42) is in close contact with the first test tube (9); completing the third layer of sealing.

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