PCR amplification device and PCR amplification method
By designing a PCR amplification device including two fixed thermal chambers and cooling parts, the problem of complex structure and inability to portability of existing PCR instruments is solved, and a compact and easy-to-operate PCR amplification device is realized, which improves the convenience of use and portability.
Patent Information
- Application Number
- CN202311318570.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-10
- Publication Date
- 2025-04-11
AI Technical Summary
The existing PCR instruments have complex structures and redundant functions, which cannot be portable and flexible, and require fixed installation, which limits scenario applications.
A PCR amplification device including two fixed thermal cavity and cooling parts is designed. The DNA amplification process is realized by installing a barrel carrying a PCR tube circulating and moving in different temperature intervals.
The device is compact, simple and easy to operate, greatly improving the convenience of use and portability, and achieving flexible and efficient progress of PCR amplification reaction.
Smart Images
Figure CN119799477A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the technical field of PCR equipment, and in particular to a PCR amplification device and a PCR amplification method. Background Art
[0002] Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be seen as a special DNA replication outside the body. The biggest feature of PCR is that it can greatly increase trace amounts of DNA to meet detection needs. PCR mainly uses a thermal cycling process of heating and cooling with the assistance of a PCR instrument to carry out biochemical reactions to replicate DNA. On the one hand, existing PCR instruments usually need to be installed in a fixed position and cannot be easily moved, which limits their application scenarios. On the other hand, they have a complex structure and redundant functions. They also usually rely on a fixed power supply to maintain the operation of the equipment, making it difficult to make them simple, portable and flexible to use. Summary of the invention
[0003] The present application provides a PCR amplification device and a PCR amplification method to solve the above-mentioned technical problems.
[0004] The embodiment of the present application is implemented as follows:
[0005] A PCR amplification device includes a temperature control component and a PCR tube installation component. The temperature control component includes a first heating element, a second heating element and a cooling element. The first heating element has a first heat chamber, and the first heat chamber has a first temperature; the second heating element has a second heat chamber, and the second heat chamber has a second temperature, and the first temperature is higher than the second temperature. The PCR tube installation component is arranged on one side of the temperature control component, and includes a mounting cylinder, a PCR tube and a connecting piece; the PCR tube is arranged in the mounting cylinder, and the connecting piece is connected to the mounting cylinder, and is used to drive the mounting cylinder to move back and forth between the first heat chamber, the second heat chamber and the cooling element.
[0006] In the PCR amplification device of the present application, when the installation tube carries the PCR tube to the first heat chamber, a DNA denaturation reaction can be performed at a first temperature, and then the second heat chamber and the first heat chamber can be cyclically heated to amplify the DNA, and then the installation tube stays in the second heat chamber to complete the final extension of the DNA. Before the installation tube that has been heated to the first temperature in the first heat chamber needs to be transferred to the second heat chamber for heating, the installation tube can be driven by the connecting member to move to the cooling member, and after the cooling member cools the installation tube to the second temperature, the installation tube is driven by the connecting member to move to the second heat chamber for heating and heat preservation. By using two fixed heat chambers and cooling members, the PCR tubes are moved to two constant temperature zones and cooling zones respectively according to the established method steps, thereby realizing the entire amplification process. The entire device has a compact and simple structure and is easy to operate, which greatly improves the convenience and portability of the device.
[0007] The embodiment of the present application further provides a PCR amplification method, which is applied to the PCR amplification device of the above embodiment, and the PCR amplification method comprises:
[0008] Inject the purified product from the incoming sample into the PCR tube in the installation cylinder, and mix the purified product from the incoming sample with the reaction solution in the PCR tube;
[0009] The first heating element heats the first heat chamber to a first temperature, and the second heating element heats the second heat chamber to a second temperature;
[0010] The installation tube moves to the first heat chamber for reaction, and after staying in the first heat chamber for a first preset time, moves to the position of the cooling element, and the cooling element cools the installation tube to a second temperature, and then moves to the second heat chamber for reaction; after staying in the second heat chamber for a second preset time, the installation tube moves to the first heat chamber again, and repeats this step;
[0011] The installation cylinder stays in the second thermal chamber for a third preset time. BRIEF DESCRIPTION OF THE DRAWINGS
[0012] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.
[0013] Figure 1 It is a schematic structural diagram of a PCR amplification device according to an embodiment of the present application.
[0014] Figure 2 for Figure 1 A schematic diagram of the structure of the PCR amplification device shown in another direction.
[0015] Figure 3 for Figure 1 Schematic diagram of the structure of the temperature control component in the PCT amplification device shown.
[0016] Figure 4 for Figure 3 Schematic diagram of the exploded structure of the temperature control component shown.
[0017] Figure 5 for Figure 1 The structure of the PCR amplification device shown is schematically shown after the base and temperature control components are removed.
[0018] Figure 6 for Figure 1 A schematic diagram of the structure of the liquid injection component in the PCR amplification device shown.
[0019] Figure 7 for Figure 1 Schematic diagram of the partial structure of the PCR amplification device shown.
[0020] Figure 8 for Figure 7 Schematic diagram of the exploded structure of the structure shown.
[0021] Fig. 9 for Figure 1 Schematic diagram of the structure of the moving components in the PCR amplification device shown.
[0022] Fig.10 for Fig. 9 A schematic diagram of the structure of the moving component shown in another direction.
[0023] Description of main component symbols:
[0024] PCR amplification device 100
[0025] Temperature control component 10
[0026] The first heating element 11 The first heat chamber 111
[0027] The first heating plate 112 The first heat conducting ring 113
[0028] The second heating element 12 The second heat chamber 121 The second heating plate 122 The second heat conducting ring 123 The first temperature sensing element 13 The second temperature sensing element 14 The first protection switch 15 The second protection switch 16 Mounting seat 17 First spacer 171 First stopper 172 The second stopper 173 Fixing 18 The second spacer 181 Cooling parts 19 Support frame 191 PCR tube installation kit 20 Installation tube 21 Cylinder 211 Locking piece 212 Hole 213 PCR tube 22 Connector 23 Cylinder 231 Push rod 232 Seal 24 Liquid injection assembly 30 Fixed seat 31 Clamping block 32 First needle tube 33 Second needle tube 34
[0029] Mobile component 40
[0030] Bottom plate 41
[0031] Guide Rail 42
[0032] Slider 421
[0033] Mounting plate 43
[0034] Connection block 431
[0035] Through slot 432
[0036] Power parts 44
[0037] Drive controller 45
[0038] Transmission parts 46
[0039] Mounting bracket 47
[0040] Protective cover 48
[0041] Displacement detection component 49
[0042] Test piece 491
[0043] Photoelectric switch 492
[0044] Detection slot 493
[0045] Base 50
[0046] Support platform 51
[0047] Positioning block 52 DETAILED DESCRIPTION
[0048] The technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all of the embodiments.
[0049] It should be noted that when an element is referred to as being "fixed to" another element, it may be directly on the other element or there may also be a centered element. When an element is considered to be "connected to" another element, it may be directly connected to the other element or there may also be a centered element. When an element is considered to be "set on" another element, it may be directly set on the other element or there may also be a centered element. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only.
[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present application. The terms used herein in the specification of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The term "or / and" used herein includes any and all combinations of one or more related listed items.
[0051] Some embodiments of the present application are described in detail. In the absence of conflict, the following embodiments and features of the embodiments can be combined with each other.
[0052] See also Figure 1 , Figure 2 and Figure 8 The present embodiment provides a PCR amplification device 100, characterized in that it includes a temperature control component 10 and a PCR tube installation component 20. The temperature control component 10 includes a first heating element 11, a second heating element 12 and a cooling element 19. The first heating element 11 has a first heat chamber 111, and the first heat chamber 111 has a first temperature. The second heating element 12 has a second heat chamber 121, and the second heat chamber 121 has a second temperature, and the first temperature is higher than the second temperature. The PCR tube installation component 20 is arranged on one side of the temperature control component 10, and includes a mounting cylinder 21, a PCR tube 22 and a connecting member 23. The PCR tube 22 is arranged in the mounting cylinder 21. The connecting member 23 is connected to the mounting cylinder 21, and is used to drive the mounting cylinder 21 to move back and forth between the first heat chamber 111, the second heat chamber 121 and the cooling element 19.
[0053] In the PCR amplification device 100 of the present application, when the installation barrel 21 carries the PCR tube 22 and moves to the first heat chamber 111, a DNA denaturation reaction can be performed at a first temperature, and then the second heat chamber 121 and the first heat chamber 111 can be cyclically heated to perform primer annealing to achieve DNA amplification, and then the installation barrel 21 stays in the second heat chamber 121 to perform a DNA polymerization reaction to complete the final extension of the DNA. Before the installation barrel 21 that has been heated to the first temperature in the first heat chamber 111 needs to be transferred to the second heat chamber 121 for heating, the connection member 23 can first drive the installation barrel 21 to move to the cooling member 19, and after the cooling member 19 cools the installation barrel 21 to the second temperature, the connection member 23 can drive the installation barrel 21 to move to the second heat chamber 121 for heating and heat preservation. By using two fixed heat chambers and cooling components 19, the PCR tube 22 is moved to two heating zones and cooling zones respectively according to the established method steps, thereby realizing the entire amplification process, without using three heating zones to meet the temperature requirements of DNA denaturation, primer annealing and DNA polymerization in the PCR amplification reaction. The entire device has a compact and simple structure and is easy to operate, which greatly improves the convenience and portability of the device.
[0054] In a specific embodiment, the first heating element 11 and the second heating element 12 are arranged in parallel, and the cooling element 19 is arranged on the side of the second heating element 12 away from the first heating element 11, so that the installation tube 21 can reciprocate between the first heating element 11, the second heating element 12 and the cooling element 19 by performing linear motion. In another embodiment, the cooling element 19 can also be arranged on the side of the first heating element 11 away from the second heating element 12, and the movement path of the installation tube 21 can be adjusted according to the setting positions of the first heating element 11, the second heating element 12 and the cooling element 19.
[0055] In one embodiment of the present application, the connecting member 23 may be a transmission structure for connecting a power device such as a motor, which supports the mounting tube 21 and can transmit the power output by the power device to the mounting tube 21, thereby driving the mounting tube 21 to move. In other embodiments, the connecting member 23 may also be a power structure such as a cylinder, a hydraulic mechanism, or a motor, which is used to output power to the mounting tube 21 to drive the mounting tube 21 to move.
[0056] Furthermore, both ends of the first heat chamber 111 are provided with openings, both ends of the second heat chamber 121 are provided with openings, and the first heat chamber 111 and the second heat chamber 121 are coaxial, so that the installation tube 21 can smoothly reciprocate in the first heat chamber 111 and the second heat chamber 121.
[0057] In one embodiment, the first heating element 11 and the second heating element 12 can be electric heating devices, and the electric heating method is resistance heating. The first heating element 11 and the second heating element 12 can be connected to a fixed power source for use, and can also be connected to a mobile power source for use. Compared with the heating element using the Peltier effect, the first heating element 11 and the second heating element 12 of the present application can reduce the use restrictions of the device, improve the flexibility and versatility of use, and can also reduce the power consumption and manufacturing cost of the device. In other embodiments, the electric heating method of the first heating element 11 and the second heating element 12 can also be electromagnetic heating or infrared heating, which can meet the heating requirements and the use requirements of ordinary power supplies, and the present application is not limited to this.
[0058] Please continue reading Figure 3 and Figure 4 In a specific embodiment, the first heating element 11 includes a first heating plate 112 and a first heat-conducting ring 113. The first heat-conducting ring 113 includes but is not limited to a structure with good thermal conductivity such as a heat-conducting copper ring. The first heating plate 112 is arranged on the surface of the first heat-conducting ring 113, and is used to heat the first heat-conducting ring 113. Specifically, the first heat-conducting ring 113 is roughly a hollow cylindrical structure, and the first heat cavity 111 is located in the first heat-conducting ring 113. The first heat cavity 111 is also roughly a cylindrical chamber with openings at both ends. The cylindrical chamber is conducive to maintaining the temperature consistency in the heat cavity, and the openings at both ends can facilitate the movement of the mounting tube 21. The first heating plate 112 is arranged around the outer peripheral surface of the first heat-conducting ring 113. In a specific embodiment, the first heating plate 112 can be in direct contact with the outer peripheral surface of the first heat-conducting ring 113 to conduct heat to the first heat-conducting ring 113. In another embodiment, a heat conducting medium may be disposed between the first heating plate 112 and the first heat conducting ring 113 to conduct heat to the first heat conducting ring 113 by indirect contact. The heat conducting medium may be heat conducting glue, heat conducting silicone grease, etc.
[0059] Furthermore, along the axial direction of the first heat-conducting ring 113, the two ends of the first heat-conducting ring 113 can be exposed from the two ends of the first heating plate 112, so as to adjust the installation position of the first heating plate 112 on the outer peripheral surface of the first heat-conducting ring 113, so that the first heating plate 112 is in the center as much as possible, and the first heat-conducting ring 113 can be evenly heated. In a specific embodiment, the first heating plate 112 is an electric heating plate, and the heating method is resistance heating. It can generate heat when powered on, and conduct the heat to the first heat-conducting ring 113 by heat conduction, thereby increasing the temperature of the first heat chamber 111. In other embodiments, the heating method of the first heating plate 112 can also be electromagnetic heating or infrared heating.
[0060] In another embodiment, the first heating plate 112 may also be disposed on the end surface of the first heat-conducting ring 113 along the axial direction of the first heat-conducting ring 113. Furthermore, there may be two first heating plates 112, which are disposed at both ends of the first heat-conducting ring 113, respectively, so as to heat different ends of the first heat-conducting ring 113 at the same time, which can improve the heating efficiency on the one hand, and evenly distribute the temperature of each part of the first heat-conducting ring 113 on the other hand, and maintain the temperature consistency of each part of the first heat-conducting ring 113.
[0061] In a specific embodiment, the second heating element 12 may also include a second heating plate 122 and a second heat-conducting ring 123, and the second heat-conducting ring 123 is coaxially arranged with the first heat-conducting ring 113. The arrangement structure of the second heating plate 122 and the second heat-conducting ring 123 is substantially the same as the arrangement structure of the first heating plate 112 and the second heat-conducting ring 123 in the aforementioned embodiments, and will not be described in detail here.
[0062] Furthermore, the cooling member 19 is arranged on the side of the second heat-conducting ring 123 away from the first heat-conducting ring 113, and is located above the second heat-conducting ring 123, and a heat dissipation temperature zone is formed at the port of the second heat-conducting ring 123 away from the first heat-conducting ring 113, so that the heat dissipation temperature zone is arranged in parallel with the second heat chamber 121 and the first heat chamber 111, and the installation tube 21 can switch between the three areas by linear motion, which is relatively simple and easy to control. In an optional embodiment, the temperature in the first heat chamber 111 and the second heat chamber 121 can be adjusted by controlling the current, resistance or voltage of the first heating plate 112 and the second heating plate 122, and the temperature of the first heat chamber 111 and the second heat chamber 121 is stably maintained at the first temperature and the second temperature, respectively. In an optional embodiment, the first temperature is 85-95°C, including but not limited to 85°C, 86°C, 87°C, 88°C, 89°C, 90°C, 91°C, 92°C, 93°C, 94°C, and 95°C; the second temperature is approximately 50-60°C, including but not limited to 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, and 60°C.
[0063] In an optional embodiment, the temperature control temperatures of the first heating element 11 and the second heating element 12 are always maintained in a constant temperature mode, the temperature error between the first heating plate 112 and the second heating plate 122 is small, the temperature is more uniform, and PID control (proportional-integral-derivative control) can be adopted to reduce the impulse of the temperature rise and fall process and maintain the temperature stability in the heat chamber.
[0064] Furthermore, the temperature control assembly 10 also includes a first temperature sensing element 13, a second temperature sensing element 14, a first protection switch 15 and a second protection switch 16. The first temperature sensing element 13 is arranged on the first heating plate 112 or the first heat-conducting ring 113, and the first protection switch 15 is arranged on the first heating plate 112. The first temperature sensing element 13 is used to detect the real-time temperature of the first heat chamber 111. When the temperature of the first heat chamber 111 exceeds the set value, the first protection switch 15 is used to disconnect the first heating plate 112, thereby achieving the function of safety protection. The second temperature sensing element 14 is arranged on the second heating plate 122 or the second heat-conducting ring 123, and the second protection switch 16 is arranged on the second heating plate 122. The second temperature sensing element 14 is used to detect the real-time temperature of the second heat chamber 121. When the temperature of the second heat chamber 121 exceeds the set value, the second protection switch 16 is used to disconnect the second heating plate 122. The disconnection methods of the first heating plate 112 and the second heating plate 122 include but are not limited to disconnecting the current, generating a circuit open, etc.
[0065] Further, the temperature control assembly 10 also includes a mounting seat 17 and a fixing member 18, and the first heating member 11 and the second heating member 12 are sandwiched between the mounting seat 17 and the fixing member 18 to fix the first heating member 11 and the second heating member 12. A first isolating member 171 is convexly provided on the side of the mounting seat 17 facing the fixing member 18, and a second isolating member 181 corresponding to the first isolating member 171 is convexly provided on the fixing member 18, and the second isolating member 181 is detachably connected to the first isolating member 171. Part of the first heating member 11 and part of the second heating member 12 are respectively arranged on opposite sides of the first isolating member 171. Another part of the first heating member 11 and another part of the second heating member 12 are respectively arranged on opposite sides of the second isolating member 181. In an optional embodiment, the mounting seat 17 and the fixing member 18 are both heat insulating members to prevent the heat of the first heating member 11 from being transferred to the second heating member 12, thereby affecting the temperature of the second heat chamber 121.
[0066] Furthermore, along the moving direction of the mounting tube 21, the mounting seat 17 is provided with a first stopper 172 and a second stopper 173 on opposite sides. The first heating element 11 is positioned between the first stopper 172 and the first isolating element 171, and the second heating element 12 is positioned between the second stopper 173 and the first isolating element 171.
[0067] In an optional embodiment, the cooling member 19 includes a fan, and a support frame 191 is provided on the side of the fixing member 18 away from the mounting seat 17. The support frame 191 partially protrudes from the side of the fixing member 18, and the fan is fixedly mounted on the support frame 191. The first heating member 11, the second heating member 12, and the fan can be fixed together by the mounting seat 17, the fixing member 18, and the support frame 191, and the entire temperature control assembly 10 can be disassembled and assembled as a whole, which can improve the assembly efficiency and facilitate the later maintenance and replacement of the device.
[0068] Please continue reading Figure 5 In one embodiment, the PCR amplification device 100 further includes a liquid injection component 30 and a moving component 40. The liquid injection component 30 is disposed on a side of the temperature control component 10 away from the PCR tube installation component 20, and is used to inject liquid into the PCR tube 22 in the installation cylinder 21. The moving component 40 is used to drive the PCR tube installation component 20 to penetrate the temperature control component 10 and move to the liquid injection component 30, so that the liquid injection component 30 can inject liquid into the PCR tube 22.
[0069] In an optional embodiment, the prepared reaction solution can be automatically injected into the PCR tube 22 by the injection assembly 30. In another optional embodiment, the reaction solution can also be automatically injected and packaged into the PCR tube 22 by an external injection device before the PCR tube 22 is loaded into the installation cylinder 21.
[0070] Furthermore, when the reaction solution is stored in the PCR tube 22, the mounting cylinder 21 can be moved to the injection assembly 30, and the injection assembly 30 automatically pushes the purified product into the PCR tube 22 to mix it with the reaction solution in the PCR tube 22, and then the PCR amplification process can be carried out.
[0071] Furthermore, the liquid injection component 30 and the temperature control component 10 are both arranged on the bottom plate 41 of the moving component 40, thereby improving the integration and structural compactness of the PCR amplification device 100 and reducing the overall size of the device.
[0072] Please continue reading Figure 6 In a specific embodiment, the liquid injection assembly 30 includes a fixed seat 31, a clamping block 32, a first needle tube 33 and a second needle tube 34. The fixed seat 31 is used to fix the installation position of the liquid injection assembly 30, and can be connected to the bottom plate 41 of the moving assembly 40 by fasteners such as screws. The clamping block 32 is installed on one side of the fixed seat 31, and the first needle tube 33 and the second needle tube 34 are clamped between the fixed seat 31 and the clamping block 32, and the first needle tube 33 and the second needle tube 34 are arranged corresponding to the first heat chamber 111. When the mounting cylinder 21 moves to the liquid injection assembly 30, the first needle tube 33 is used to inject liquid into the PCR tube 22, and the second needle tube 34 is used to discharge the gas in the PCR tube 22 during the liquid injection process. Further, along the direction toward the temperature control assembly 10, the end of the first needle tube 33 exceeds the second needle tube 34, so that the first needle tube 33 can start to inject liquid from the bottom of the PCR tube 22, and the gas in the liquid squeeze tube is discharged from the second needle tube 34, reducing the problem of liquid leakage.
[0073] Please continue reading Figure 7 and Figure 8In an optional embodiment, the installation cylinder 21 includes a cylinder body 211 and a locking member 212, and the PCR tube 22 is inserted into the inner cavity of the cylinder body 211. The locking member 212 is covered at the end of the PCR tube 22 and is threadedly connected to the cylinder body 211 to fix the PCR tube 22 into the cylinder body 211. A sealing member 24 is also provided in the installation cylinder 21, and the sealing member 24 includes but is not limited to a sealing rubber plug and has a certain elasticity. The sealing member 24 blocks the port of the PCR tube 22 and is clamped between the locking member 212 and the PCR tube 22. A hole 213 is provided on the locking member 212 of the installation cylinder 21, and the hole 213 is coaxial with the cylinder body 211 and corresponds to the first needle tube 33 and the second needle tube 34 of the injection assembly 30. The top of a part of the sealing member 24 can be exposed from the hole 213, so that the first needle tube 33 and the second needle tube 34 can pierce the sealing member 24 from the hole 213 and communicate with the PCR tube 22. The seal 24 is made of elastic material. After the first needle tube 33 and the second needle tube 34 are removed from the PCR tube 22, the seal 24 can automatically block the pinhole 213 produced by the puncture of the first needle tube 33 and the second needle tube 34 to prevent leakage of the liquid in the PCR tube 22. In one embodiment, the barrel 211 and the locking member 212 of the mounting barrel 21 are made of heat-conducting material so that the mounting barrel 21 can transfer the heat in the first heat chamber 111 or the second heat chamber 121 to the PCR tube 22. The heat-conducting material includes but is not limited to aluminum, copper, heat-conducting silicone, etc. A temperature detector can also be set on the mounting barrel 21 to monitor the temperature of the PCR tube 22 in real time.
[0074] Please continue reading Fig. 9 and Fig.10 In an optional embodiment, the moving assembly 40 includes a bottom plate 41, a guide rail 42, a mounting plate 43, and a power member 44. The guide rail 42 is fixed to the bottom plate 41, the mounting plate 43 is movably connected to the guide rail 42, the PCR tube mounting assembly 20 is connected to the mounting plate 43, and the liquid injection assembly 30, the temperature control assembly 10 and the power member 44 are all arranged on the bottom plate 41. The power member 44 is transmission-connected to the mounting plate 43, and the power member 44 is used to drive the mounting plate 43 to move along the guide rail 42, thereby driving the PCR tube mounting assembly 20 to move to the liquid injection assembly 30 or the temperature control assembly 10.
[0075] In an optional embodiment, the liquid injection component 30 and the temperature control component 10 are spaced and fixedly arranged on the same side of the bottom plate 41, and the guide rail 42 is fixed on the side of the bottom plate 41 away from the liquid injection component 30 and the temperature control component 10. A through slot 432 is provided on the mounting plate 43, and the bottom plate 41 can pass through the through slot 432, so that the mounting plate 43 is sleeved on one end of the bottom plate 41, and the bottom plate 41 and the inner side wall of the through slot 432 are spaced apart, so that the mounting plate 43 can move relative to the bottom plate 41. A slider 421 is slidably arranged on the guide rail 42, and a connecting block 431 corresponding to the slider 421 is provided on the mounting plate 43. The connecting block 431 is fixedly connected to the slider 421. When the power member 44 drives the mounting plate 43 to move toward the liquid injection component 30 or the temperature control component 10, the slider 421 moves along the guide rail 42 driven by the mounting plate 43, thereby guiding the moving direction of the mounting plate 43 and improving the movement stability.
[0076] In an optional embodiment, the connecting member 23 includes a cylinder 231 and a push rod 232. The cylinder 231 is fixed on the side of the mounting plate 43 facing the temperature control component 10. The push rod 232 connects the mounting tube 21 and the cylinder 231. When the power member 44 drives the mounting plate 43 to move the connecting member 23 and the mounting tube 21 to the position of the temperature control component 10, the cylinder 231 drives the push rod 232 to extend and retract to drive the mounting tube 21 to move in a small range between the first heat chamber 111, the second heat chamber 121 and the cooling component 19 of the temperature control component 10, thereby reducing the vibration generated by the entire moving component 40 during the movement.
[0077] Further, the power member 44 is fixed on the base plate 41 by a mounting bracket 47, and the power member 44 can be but is not limited to a drive motor. The moving assembly 40 also includes a drive controller 45, which is arranged on the base plate 41 and electrically connected to the power member 44. The drive controller 45 is used to control the operation of the power member 44. The drive controller 45 can also be connected to an external control terminal in communication to receive a control signal from the external control terminal, so as to adjust the operation mode of the power member 44. A transmission member 46 is connected between the power member 44 and the mounting plate 43, and the power member 44 drives the mounting plate 43 to move along the guide rail 42 through the transmission member 46. In one embodiment, the transmission member 46 is a screw rod, and the screw rod is threadedly connected to the mounting plate 43. When the power member 44 drives the screw rod to rotate, it drives the mounting plate 43 to move along the screw rod. The moving distance of the mounting plate 43 can be accurately controlled by the number of turns of the screw rod, and the motion state is stable and not prone to vibration. When the connecting member 23 is used as a supporting structure, the mounting cylinder 21 can move synchronously with the mounting plate 43, and the mounting cylinder 21 can be driven to reciprocate between the first heat chamber 111, the second heat chamber 121 and the cooling member 19 by the power member 44 and the transmission member 46. In other embodiments, the transmission member 46 can also be a belt transmission mechanism, so that the power member 44 and the mounting plate 43 can be connected by transmission through structures such as synchronous wheels and synchronous belts. When the power member 44 drives the synchronous belt to rotate through the synchronous wheel, the mounting plate 43 is pulled, so that the mounting plate 43 moves along the guide rail 42.
[0078] Further, please refer again to Figure 1 and Figure 5 , the moving assembly 40 also includes a protective cover 48 and a displacement detection assembly 49. The protective cover 48 is arranged at the bottom of the base plate 41, and the guide rail 42 and part of the mounting plate 43 are accommodated in the protective cover 48. The displacement detection assembly 49 includes a detection piece 491 and a photoelectric switch 492. The detection piece 491 is fixedly arranged on one side of the mounting plate 43, and the photoelectric switch 492 is arranged on one side of the protective cover 48 and corresponds to the detection piece 491. The photoelectric switch 492 is provided with a detection slot 493. When the mounting plate 43 is in the zero position, the detection piece 491 is partially accommodated in the detection slot 493, triggering the photoelectric switch 492 to send a zero position signal of the mounting plate 43. The drive controller 45 can be connected to the photoelectric switch 492 for communication, so as to judge the position of the mounting plate 43 according to the signal of the photoelectric switch 492, and control the operation of the power part 44.
[0079] In an alternative embodiment, please refer again to Figure 1 and Figure 2The PCR amplification device 100 further includes a base 50, which is disposed at the bottom of the moving component 40 and is detachably connected to the protective cover 48, and is used to connect the PCR amplification device 100 to other devices, so as to facilitate the overall disassembly and assembly of the PCR amplification device 100. The base 50 includes a positioning block 52 and a support platform 51, and one or more positioning blocks 52 are detachably disposed on the surface of the support platform 51, and the protective cover 48 of the moving component 40 is fixedly connected to the positioning block 52.
[0080] Furthermore, after the PCR amplification device 100 completes the PCR amplification reaction, the amplification product can be automatically pushed into a downstream PCR purification device for processing by a syringe pump or other external pneumatic automation device.
[0081] An optional embodiment further provides a PCR amplification method, which is applied to the PCR amplification device 100 of the above embodiment, and the PCR amplification method includes:
[0082] Inject the purified product from the incoming sample into the PCR tube 22 in the installation cylinder 21, and mix the purified product from the incoming sample with the reaction solution in the PCR tube 22;
[0083] The first heating element 11 heats the first heat chamber 111 to a first temperature, and the second heating element 12 heats the second heat chamber 121 to a second temperature;
[0084] The installation tube 21 moves to the first heat chamber 111 for reaction. After staying in the first heat chamber 111 for a first preset time, the installation tube 21 moves to the position of the cooling element 19. The cooling element 19 cools the installation tube 21 to a second temperature. The installation tube 21 moves to the second heat chamber 121 for reaction. After staying in the second heat chamber 121 for a second preset time, the installation tube 21 moves to the first heat chamber 111 again, and this step is repeated.
[0085] The installation cylinder 21 stays in the second heat chamber 121 for a third preset time.
[0086] Furthermore, the third preset time is greater than the second preset time.
[0087] In an optional embodiment, the step of “injecting the purified product from the future sample into the PCR tube 22 in the mounting barrel 21 ” includes: injecting the purified product from the future sample into the PCR tube 22 via the liquid injection assembly 30 .
[0088] Specifically, before the purified product is injected into the PCR tube 22, when the reaction solution (such as a mix reagent) is already encapsulated in the PCR tube 22, the installation tube 21 of the PCR tube installation assembly 20 carries the PCR tube 22 to the injection assembly 30, and the first needle tube 33 and the second needle tube 34 of the injection assembly 30 pass through the locking member 212 and the sealing member 24 and enter the inner cavity of the PCR tube 22. The first needle tube 33 exceeds the second needle tube 34, so that the first needle tube 33 can be inserted deeper into the PCR tube 22. The purified product can be automatically injected into the PCR tube 22 through the first needle tube 33 by an external pneumatic device, and mixed with the reaction solution already loaded in the PCR tube 22. At the same time, the second needle tube 34 exhausts and releases the pressure of the PCR tube 22 to avoid liquid leakage caused by excessive pressure in the tube. The method of pre-storing the reaction solution in the PCR tube is as described in the previous embodiment, and will not be repeated here.
[0089] In another embodiment, the step of “injecting the purified product from the incoming sample into the PCR tube 22 in the mounting barrel 21 ” includes: injecting the reaction solution and the purified product from the incoming sample into the PCR tube 22 through the injection assembly 30 , respectively.
[0090] Specifically, the installation barrel 21 carries the empty PCR tube 22 to the injection assembly 30, and the reaction solution is injected into the PCR tube 22 through the first needle tube 33, and the injection method is the same as above. Then, the installation barrel 21 carries the PCR tube 22 filled with the reaction solution and is separated from the injection assembly 30. When the injection assembly 30 is ready to inject the incoming sample purification product, the installation barrel 21 carries the PCR tube 22 to the injection assembly 30 again, and injects the incoming sample purification product into the PCR tube 22 through the first needle tube 33, so that the incoming sample purification product is mixed with the reaction solution.
[0091] Further, after the reaction liquid in the PCR tube 22 is mixed with the sample purification product, the moving assembly 40 will drive the PCR tube installation assembly 20 to move backward, so that the PCR tube 22 leaves the area of the temperature control assembly 10. At this time, the temperature control assembly 10 starts to work, the first heating plate 112 and the second heating plate 122 are powered on and heat the first heat-conducting ring 113 and the second heat-conducting ring 123 respectively, so that the first heat cavity 111 in the first heat-conducting ring 113 is maintained at the first temperature, and the second heat cavity 121 in the second heat-conducting ring 123 is maintained at the second temperature. In an optional embodiment, the first temperature is 90°C and the second temperature is 55°C.
[0092] Next, the mounting plate 43 of the moving assembly 40 drives the PCR tube mounting assembly 20 back to the set zero position. When the photoelectric switch 492 recognizes that the detection piece 491 on the mounting plate 43 is in place, the guide rail 42, the drive controller 45, the power member 44, the transmission member 46 and the mounting plate 43 and other components work together to move the mounting cylinder 21 of the PCR tube mounting assembly 20 to the 90°C temperature zone for reaction. After staying in the temperature zone for the first preset time, the connector 23 of the PCR tube mounting assembly 20 will move the mounting cylinder 21 to the position of the cooling member 19. The cooling member 19 starts to work to cool the PCR tube 22. When the temperature of the PCR tube 22 drops to 55°C, the cooling member 19 stops working, and the connector 23 moves the mounting cylinder 21 to the 55°C temperature zone for reaction, and stays in the temperature zone for the second preset time. After the reaction at 55°C is completed, the connector 23 continues to move the mounting cylinder 21 to the 90°C temperature zone, and repeats the above steps. In an optional embodiment, the first preset time is about 10s, and the second preset time is about 90s.
[0093] In an exemplary embodiment, the mixed liquid in the PCR tube 22 undergoes thirty cycles of unwinding, binding and extension at 90°C for 10s and 55°C for 90s to complete the amplification of the DNA sample. When the cycle is completed, if the PCR tube 22 is in the 55°C temperature zone, the PCR tube 22 will continue to stay in the 55°C temperature zone for the third preset time for the final extension, so that the DNA single strand is completely restored to a double strand, completing the PCR reaction process. If the PCR tube 22 is in the 90°C temperature zone when the cycle is completed, the connector 23 will first drive the PCR tube 22 to move to the cooling member 19, and after the PCR tube 22 is cooled to 55°C, it will move to the 55°C temperature zone and stay for the third preset time for the final extension. In an optional embodiment, the third preset time is approximately 600s.
[0094] In an exemplary experimental process, the injection process of the sample purification product was completed in 50 seconds, and the PCR amplification process took about 130 minutes. The test results showed that the initial input amount was 0.1 ng / ul, and the amplification result could reach 54 ng / ul. The whole process was automated, without human intervention, and the PCR amplification efficiency was improved. The entire PCR amplification device 100 has a simple structure and is easy to operate and control, the process is simplified to the greatest extent, and the accuracy is guaranteed.
[0095] The above implementation modes are only used to illustrate the technical solutions of the present application and are not intended to limit the present application. Although the present application has been described in detail with reference to the above preferred implementation modes, a person skilled in the art should understand that the technical solutions of the present application may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present application.
Claims
1. A PCR amplification device, characterized in that: include: The temperature control component comprises a first heating element, a second heating element and a cooling element, wherein the first heating element has a first heat cavity, and the first heat cavity has a first temperature; The second heating element has a second heat chamber, the second heat chamber has a second temperature, and the first temperature is higher than the second temperature; A PCR tube installation assembly is movably arranged on one side of the temperature control assembly, and includes a mounting cylinder, a PCR tube and a connector; The PCR tube is arranged in the installation cylinder; the connecting member is connected to the installation cylinder, and is used to drive the installation cylinder to move back and forth between the first heat chamber, the second heat chamber and the cooling member.
2. The PCR amplification device according to claim 1, characterized in that: The first heating element and the second heating element are arranged in parallel, both ends of the first heat chamber are provided with openings, both ends of the second heat chamber are provided with openings, and the first heat chamber and the second heat chamber are coaxial.
3. The PCR amplification device according to claim 1, characterized in that: The temperature-reducing component is disposed on a side of the second heating component facing away from the first heating component; or, the temperature-reducing component is disposed on a side of the first heating component facing away from the second heating component.
4. The PCR amplification device according to claim 1, characterized in that: The first heating element comprises a first heating plate and a first heat-conducting ring, the first heat cavity is arranged in the first heat-conducting ring, and the first heating plate is arranged on the surface of the first heat-conducting ring for heating the first heat-conducting ring; and / or The second heating element includes a second heating plate and a second heat-conducting ring. The second heat cavity is disposed in the second heat-conducting ring. The second heating plate is disposed on the surface of the second heat-conducting ring for heating the second heat-conducting ring.
5. The PCR amplification device according to claim 4, characterized in that: The temperature control assembly includes a first temperature sensing component and a first protection switch, wherein the first temperature sensing component is disposed on the first heating plate or the first heat-conducting ring, and the first protection switch is disposed on the first heating plate; The first temperature sensing element is used to monitor the real-time temperature of the first heat chamber. When the temperature of the first heat chamber exceeds a set value, the first protection switch is used to disconnect the first heating plate. and / or The temperature control component includes a second temperature sensing component and a second protection switch. The second temperature sensing component is arranged on the second heating plate or the second heat-conducting ring, and the second protection switch is arranged on the second heating plate. The second temperature sensing component is used to monitor the real-time temperature of the second heat chamber. When the temperature of the second heat chamber exceeds a set value, the second protection switch is used to disconnect the second heating plate.
6. The PCR amplification device according to claim 1, characterized in that: The temperature control component also includes a mounting seat and a fixing member, the first heating member and the second heating member are clamped between the mounting seat and the fixing member, the mounting seat is provided with a first isolating member protruding from one side facing the fixing member, the fixing member is provided with a second isolating member corresponding to the first isolating member, part of the first heating member and part of the second heating member are respectively arranged on opposite sides of the first isolating member, and another part of the first heating member and another part of the second heating member are respectively arranged on opposite sides of the second isolating member.
7. The PCR amplification device according to claim 6, characterized in that: A first stopper and a second stopper are disposed on opposite sides of the mounting base. The first heating element is positioned between the first stopper and the first isolating element. The second heating element is positioned between the second stopper and the first isolating element.
8. The PCR amplification device according to claim 6, characterized in that: The cooling component comprises a fan. A support frame is provided on the side of the fixing component facing away from the mounting seat. The support frame partially protrudes from the side of the fixing component. The fan is fixedly mounted on the support frame.
9. The PCR amplification device according to claim 1, characterized in that: The PCR amplification device also includes a liquid injection component, which is arranged on a side of the temperature control component away from the PCR tube installation component and is used to inject liquid into the PCR tube in the installation cylinder.
10. The PCR amplification device according to claim 9, characterized in that: The injection assembly includes a fixed seat, a clamping block, a first needle tube and a second needle tube. The clamping block is installed on one side of the fixed seat. The first needle tube and the second needle tube are clamped between the fixed seat and the clamping block. Along the direction toward the temperature control assembly, the end of the first needle tube exceeds the end of the second needle tube.
11. The PCR amplification device according to claim 1, characterized in that: The installation cylinder includes a cylinder body and a locking piece, and the PCR tube is inserted into the inner cavity of the cylinder body; the locking piece is covered at the end of the PCR tube and connected to the cylinder body, and a sealing piece is also provided in the installation cylinder, which blocks the port of the PCR tube and is clamped between the locking piece and the PCR tube; a hole is opened on the locking piece, and part of the sealing piece is exposed from the hole.
12. The PCR amplification device according to claim 1, characterized in that: The PCR amplification device also includes a moving component, which includes a base plate, a guide rail, a mounting plate, and a power component. The guide rail is fixed to the base plate, the mounting plate is movably connected to the guide rail, the connecting component connects the mounting plate and the mounting cylinder, the temperature control component and the power component are both arranged on the base plate, the power component is transmission-connected to the mounting plate, and the power component is used to drive the mounting plate to move along the guide rail, thereby driving the PCR tube mounting component to move to the temperature control component.
13. The PCR amplification device according to claim 12, characterized in that: The guide rail is fixed to a side of the base plate away from the temperature control component, a through slot is provided on the mounting plate, the base plate passes through the through slot, a slider is slidably arranged on the guide rail, a connecting block is provided on the mounting plate, and the connecting block is fixedly connected to the slider.
14. A PCR amplification method, applied to the PCR amplification device according to any one of claims 1 to 13, characterized in that: The PCR amplification method comprises: Inject the purified product from the incoming sample into the PCR tube in the installation cylinder, and mix the purified product from the incoming sample with the reaction solution in the PCR tube; The first heating element heats the first heat chamber to a first temperature, and the second heating element heats the second heat chamber to a second temperature; The installation tube moves to the first heat chamber for reaction, and after staying in the first heat chamber for a first preset time, moves to the position of the cooling element, and the cooling element cools the installation tube to a second temperature, and then moves to the second heat chamber for reaction; after staying in the second heat chamber for a second preset time, the installation tube moves to the first heat chamber again, and repeats this step; The installation cylinder stays in the second thermal chamber for a third preset time.