Nucleic acid amplification system based on double-temperature-zone control

Abstract

The invention relates to a nucleic acid amplification system based on double temperature zone control, the system is composed of a motor control system, a reaction tank and a double temperature control module, the reaction tank is precisely driven by a stepping motor to move in an independent temperature zone, and efficient conversion and uniform temperature conduction of nucleic acid denaturation, annealing and extension processes at different set temperatures are realized. Compared with the prior art, the method has the advantages that the amplification efficiency and precision are improved, and the influence of temperature gradient on the amplification process is reduced.

Description

Technical Field

[0001] The present invention relates to the technical fields of molecular biology and biomedical detection, and particularly to a nucleic acid amplification system based on dual-temperature zone control. Background Art

[0002] Nucleic acid amplification technology is an important tool in molecular biology and clinical diagnosis, and is widely used in fields such as pathogen detection, genetic disease screening, and gene mutation analysis. Currently, PCR (Polymerase Chain Reaction) is the most commonly used nucleic acid amplification method, and usually completes the denaturation, annealing, and extension steps by raising and lowering the temperature of a metal heating block. However, traditional PCR systems have problems such as slow temperature conversion rate, high energy consumption, and unstable temperature control, resulting in a longer amplification time and affecting the detection efficiency.

[0003] In recent years, fixed-temperature zone amplification technology has been proposed, that is, the reaction reagent is moved between different temperature zones mechanically to improve the amplification efficiency. However, existing fixed-temperature zone systems still have problems such as low mechanical control accuracy, poor temperature control stability, and insufficient automation, and it is difficult to meet the requirements of efficient and accurate nucleic acid detection. For this reason, the present invention provides a nucleic acid amplification system based on dual-temperature zone control, which uses a stepper motor to drive the reaction tank to move between two temperature zones, and precisely adjusts the temperature through a thermostat, thereby improving the speed and stability of nucleic acid amplification, simplifying the operation process, and enhancing the automation degree of the system. Summary of the Invention

[0004] The purpose of the present invention is to overcome the disadvantages of existing PCR nucleic acid amplification technologies, such as slow temperature conversion rate, long amplification time, complex system, and low automation degree, and provide a nucleic acid amplification system based on dual-temperature zone control.

[0005] The purpose of the present invention can be achieved by the following technical solutions:

[0006] A nucleic acid amplification system based on dual-temperature zone control, comprising a motor control system, a reaction tank, and a temperature control module. Among them,

[0007] The motor control system accurately drives the reaction tank to move between two temperature zones through a stepper motor, and the positioning accuracy is ≤10 μm, ensuring that each stage of the nucleic acid amplification process can be completed at an accurate temperature. The motor control system is connected to the stepper motor through a control circuit, and can precisely adjust the rotation angle of the stepper motor to achieve precise movement of the reaction tank between different temperature zones.

[0008] The reaction tank has a high - heat - conduction geometric structure, which can enhance temperature uniformity, reduce temperature difference, and improve amplification efficiency. The reaction tank is provided with a square groove and multiple pores. The pore wall thickness is 0.2 - 0.5 mm, which is used to place samples and reagents, ensuring good contact between the sample and the temperature zone during the reaction process, thereby enhancing the efficiency of nucleic acid amplification.

[0009] The temperature control module includes two independent temperature - controlled areas, which are respectively used for the denaturation, annealing, and extension processes of nucleic acid amplification. Each temperature - controlled area is controlled by an independent thermostat. The temperature is adjusted in real - time through the thermostat to ensure that the temperature within the temperature zone is always maintained within the set range, so as to improve amplification accuracy.

[0010] According to the above - mentioned technical solution, the motor control system of the system can make the reaction tank move smoothly between the two temperature zones through the precise drive of the stepper motor. Within each temperature zone, the temperature control module will automatically adjust the temperature of the temperature zone to ensure temperature stability during the nucleic acid amplification process.

[0011] Furthermore, the square groove designed in the reaction tank can make the temperature distribution between the reaction sample and the tank wall more uniform, avoiding the temperature difference problem that may occur in traditional circular reaction tanks. This design ensures that denaturation, annealing, and extension in all stages of nucleic acid amplification can be carried out under appropriate temperature conditions, improving the efficiency of the amplification reaction.

[0012] Furthermore, the temperature control module includes two independent temperature - controlled areas, which are respectively used for the denaturation, annealing, and extension stages during nucleic acid amplification. The temperature in each temperature zone is precisely adjusted by the thermostat to ensure that the temperature of each temperature zone is always within the set range, optimizing the temperature stability and accuracy in the nucleic acid amplification reaction.

[0013] Furthermore, the thermostat can monitor the temperature in real - time through the feedback control system and adjust the power of the heating device according to the set temperature requirements to maintain the required temperature. By precisely controlling the temperature of each temperature zone, strict control of the temperature in each stage during the amplification process is ensured, thereby improving the amplification efficiency and accuracy.

[0014] Furthermore, the motor control system and the temperature control module cooperate with each other, and can automatically switch temperature zones during nucleic acid amplification to adapt to the temperature requirements of different stages. The motor control system ensures that the reaction tank can switch to different temperature zones at the appropriate time, and the temperature control module ensures that the temperature of each stage is accurate.

[0015] Furthermore, the system can realize automatic control of the entire nucleic acid amplification process through a simplified operation interface and an automated control system. Users only need to set the reaction temperature and time, and the system will automatically perform temperature control and temperature - zone switching, greatly simplifying the operation process.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention provides a nucleic acid amplification system based on dual-temperature zone control. By driving the reaction tank to move between two temperature zones with a stepper motor and combining with a thermostat for precise temperature control, an efficient, precise, and automated nucleic acid amplification process is achieved. Compared with traditional PCR, the present invention has significantly improved amplification speed, temperature control accuracy, system automation, and stability, providing an efficient and reliable solution for rapid and precise nucleic acid detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is a schematic diagram of the overall structure of the nucleic acid amplification system based on dual-temperature zone control of the present invention;

[0019] Reference numerals:

[0020] 10 - Stepper motor, 20 - First temperature zone heating block (high-temperature denaturation zone), 30 - Reaction tank, 40 - Second temperature zone heating block (low-temperature annealing / extension zone). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] The present invention will be described in detail below with reference to the drawings and specific embodiments. This embodiment is implemented on the premise of the technical solution of the present invention, and the detailed implementation manner and specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments.

[0022] The present invention provides a nucleic acid amplification system based on dual-temperature zone control, and its schematic diagram is as Figure 1 shown.

[0023] Among them, the motor control system uses a stepper motor 10 connected to a drive circuit, and the rotation angle of the stepper motor is precisely adjusted through a control circuit to drive the reaction tank 30 to move smoothly between the first temperature zone heating block 20 and the second temperature zone heating block 40. The movement accuracy of the stepper motor can reach the micron level, ensuring the accurate positioning of the reaction tank, so as to ensure that each stage of nucleic acid amplification can be carried out at the set temperature.

[0024] Among them, the geometric structure of the reaction tank 30 is square, and this design can effectively improve the uniformity of temperature distribution. The reaction tank 30 is provided with a plurality of hole positions for placing samples and reagents. The material of the reaction tank is selected as a material with good thermal conductivity to ensure that the temperature can be evenly conducted during the reaction process and avoid the influence of uneven reaction solution temperature on the amplification effect.

[0025] Among them, the temperature control module consists of a first temperature zone heating block 20 and a second temperature zone heating block 40, corresponding to the three processes of denaturation, annealing, and extension in nucleic acid amplification respectively. Each temperature control zone is controlled by an independent thermostat, and the thermostat monitors and adjusts the temperature of each temperature zone in real time to ensure that the temperature is stable within the required range.

[0026] The reaction tank 30 is fixed to the output shaft of the stepping motor 10. The distance between the first temperature zone heating block 20 and the second temperature zone heating block 40 is 8 cm. The thermostats are set to 98 °C (high temperature zone) and 70 °C (low temperature zone). According to the experimental requirements, the nucleic acid sample to be amplified is injected into the holes of the reaction tank 30, and the temperature and time of each stage are set through the control interface. For example, the denaturation temperature is set to 98 °C and stays for 30 seconds, the annealing and extension temperature is 70 °C and stays for 45 seconds, and it cycles 40 times. The system automatically starts according to the set parameters, and the motor control system starts to drive the reaction tank 30 to move between the first temperature zone heating block 20 and the second temperature zone heating block 40. The temperature of each temperature zone is precisely adjusted by the thermostat to ensure the stability of the temperature at each stage. The system automatically completes each stage of nucleic acid amplification, including denaturation, annealing, and extension. After each stage is completed, the reaction tank 30 automatically moves to the next temperature zone to ensure the continuity and accuracy of the reaction. When all amplification stages are completed, the system automatically stops, obtains the amplification result, and conducts subsequent analysis.

[0027] The preferred specific embodiments of the present invention have been described in detail above. It should be understood that those of ordinary skill in the art can make many modifications and variations according to the concept of the present invention without creative labor. Therefore, all technical solutions that can be obtained by those skilled in the art in the technical field of the present invention based on the concept of the present invention through logical analysis, reasoning, or limited experiments on the basis of the prior art should be within the protection scope determined by the claims.

Claims

1. A nucleic acid amplification system based on dual-temperature zone control, characterized in that, The system mainly includes: a motor control system for precisely driving a reaction tank to move horizontally between two independent temperature control zones through a stepper motor; a reaction tank for accommodating nucleic acid amplification reaction reagents, fixed on the stepper motor, and the reaction tank adopts a high heat conduction structure to improve temperature uniformity and reduce the influence of temperature gradient on the amplification process; a temperature control module including two independent temperature control zones respectively for nucleic acid denaturation, annealing and extension processes, and the temperature control zones achieve precise temperature regulation through independent heating modules.

2. The nucleic acid amplification system according to claim 1, wherein The motor control system includes a stepper motor, a stepper motor driver and a control circuit. The stepper motor driver is used to receive control signals and drive the stepper motor, with a positioning accuracy ≤ 10 μm, so as to achieve precise movement of the reaction tank between different temperature control zones and ensure that the reaction process is completed according to the set parameters.

3. The nucleic acid amplification system according to claim 1, wherein The reaction tank is made of materials resistant to high temperature and chemical corrosion, including but not limited to polycarbonate (PC), polytetrafluoroethylene (PTFE) or silicone rubber, and is internally provided with a square groove and multiple pores, with the pore wall thickness of 0.2 - 0.5 mm, so as to adapt to the high temperature denaturation process and have good heat conductivity.

4. The nucleic acid amplification system according to claim 1, wherein The temperature control module includes: a first temperature zone for providing a high temperature environment (95 - 98 °C) required for the denaturation reaction; a second temperature zone for providing a lower temperature environment (65 - 72 °C) required for the annealing and extension reactions; the two temperature zones are respectively controlled by independent heating modules, and the temperature of each is adjusted through a thermostat to ensure the stability and accuracy of temperature during the amplification process.

5. The nucleic acid amplification system according to claim 1, characterized in that, The reaction tank is fixed to the stepper motor through a detachable connection method, so as to replace reaction tanks of different specifications to adapt to different experimental requirements, and move between the first temperature zone and the second temperature zone along a set path through the stepper motor to complete the denaturation, annealing and extension reactions in the nucleic acid amplification process.

6. The nucleic acid amplification system according to claim 1, wherein The system is connected to a computer terminal control software, and the user can set an amplification program on the software interface, including but not limited to parameters such as residence time in the temperature zone, moving speed, number of cycles, etc. The system automatically executes the amplification process according to the set parameters, improving the automation level and simplifying the operation.

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