Integrated microwave temperature control automatic nucleic acid amplification system and its working method

Abstract

The application relates to the field of nucleic acid amplification technology and provides a full-automatic nucleic acid amplification system integrated with microwave temperature control and a working method thereof.The system comprises a microprocessor, a temperature heating module, a nucleic acid amplification container, a microwave generator connected with the microprocessor and a thermocouple, the microwave generator is connected with the temperature heating module, the temperature heating module adopts a planar microwave resonator or a waveguide, the microwave generator is used for outputting microwave signals with specific frequency and power, the temperature heating module is used for directing the microwave signals to nucleic acid amplification reagents in the nucleic acid amplification container to heat the temperature of the nucleic acid amplification reagents, the thermocouple is used for monitoring the temperature of the nucleic acid amplification reagents and uploading the temperature to the microprocessor, and the microprocessor is used for controlling the frequency and power of the microwave signals according to the temperature of the nucleic acid amplification reagents.The temperature can be rapidly raised, the sample volume is reduced, and the nucleic acid amplification time is shortened.

Description

Technical Field

[0001] This invention relates to the field of nucleic acid amplification technology, and in particular to an integrated microwave temperature-controlled fully automated nucleic acid amplification system and its operating method. Background Technology

[0002] The statements in this section are merely background information related to the present invention and do not necessarily constitute prior art.

[0003] Nucleic acid amplification technology is widely used in life sciences, biomedicine, food safety, and other fields, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification. Both technologies achieve exponential amplification of nucleic acid templates through temperature control. Each cycle of polymerase chain reaction (PCR) includes three stages: denaturation, annealing, and extension. Different stages correspond to specific temperatures, including high-temperature denaturation, low-temperature annealing, and medium-temperature extension. For effective nucleic acid amplification, the temperature of the thermal cycling stages must be precisely controlled. Current nucleic acid amplification equipment generally uses Peltier elements for temperature control, achieving temperature control through the Peltier effect generated by excellent thermoelectric materials. Peltier elements mainly use N-type and P-type semiconductors to form a thermocouple. When current flows through the thermocouple circuit, energy transfer occurs. Depending on the direction of the current, endothermic and exothermic phenomena are exhibited at the junctions of different conductors. Therefore, precise control of nucleic acid temperature is achieved by controlling the endothermic and exothermic processes.

[0004] Currently, Peltier element-based nucleic acid amplification equipment requires the connection of a metal temperature control component. When current flows through the Peltier element, one side heats up and the other side cools down. Temperature control of the metal temperature control component is achieved by adjusting the direction and intensity of the current. The metal temperature control component transfers heat to the nucleic acid amplification reaction vessel via thermal conduction, thus achieving precise temperature control of the nucleic acid amplification reaction system. However, this method has some drawbacks:

[0005] (1) High power consumption: Peltier elements achieve temperature control by controlling the input current, which requires an external power supply. Controlling the temperature requires a lot of electrical energy, resulting in high power consumption.

[0006] (2) Low heat transfer efficiency: During the electrical energy conversion process, a portion of the electrical energy is converted into uncontrollable waste heat, requiring heat dissipation design to ensure normal equipment operation. Simultaneously, energy transfer to the nucleic acid amplification solvent relies on metal temperature components, resulting in low thermal conductivity and high thermal resistance between the heating block and the reaction tube. This inevitably leads to energy diffusion to the outside, resulting in low heat transfer efficiency. In other words, the temperature control of the nucleic acid solution depends on heat conduction, leading to slow thermal circulation, low thermal conductivity, and high thermal resistance between the heating block and the reaction tube, severely limiting the temperature control rate.

[0007] (3) Uneven temperature gradient: There is a temperature gradient on the surface of the Peltier element, the metal temperature component and the PCR container. In particular, the large size of commercial multi-PCR tubes can easily cause uneven heating and inhibit the amplification reaction. At the same time, too much energy is required to maintain temperature uniformity.

[0008] (4) Low temperature control rate: The heating rate of commercial nucleic acid amplification instruments is usually limited to 4-6℃ / s, which requires 1.5-2h to complete 30-40 cycles. When the primer annealing temperature is high and the target fragment is small, annealing and extension can be combined, simplifying the traditional three-step method into a two-step method, which shortens the nucleic acid amplification reaction time and improves the detection efficiency, but may affect enzyme efficiency.

[0009] Therefore, the development of efficient and rapid PCR detection equipment needs to meet two major requirements: small reaction volume and fast temperature control rate. Summary of the Invention

[0010] To address the shortcomings of existing technologies, this invention provides an integrated microwave temperature-controlled fully automated nucleic acid amplification system and its operating method. It utilizes a planar microwave resonator or waveguide to directionally transmit microwave signals to the nucleic acid amplification reagent, achieving rapid temperature rise. Furthermore, the use of a microwave resonator or waveguide allows for targeted heating of the liquid within a small space, reducing the volume of the heated sample. Simultaneously, a microprocessor monitors the temperature of the nucleic acid amplification solution in real time and precisely controls the sample temperature by adjusting the microwave transmitter power or frequency through feedback, thus shortening the time required for nucleic acid amplification.

[0011] To achieve the above objectives, the present invention adopts the following technical solution:

[0012] The first aspect of this invention provides an integrated microwave temperature-controlled fully automated nucleic acid amplification system.

[0013] An integrated microwave temperature-controlled fully automated nucleic acid amplification system includes a microprocessor, a temperature heating module, a nucleic acid amplification container, and a microwave generator and a thermocouple connected to the microprocessor. The microwave generator is connected to the temperature heating module, and the temperature heating module adopts a planar microwave resonator or a waveguide.

[0014] The microwave generator is used to output microwave signals of a specific frequency and power.

[0015] The temperature heating module is used to transmit microwave signals in a directional manner to the nucleic acid amplification reagent in the nucleic acid amplification container, thereby heating the nucleic acid amplification reagent to a certain temperature.

[0016] The thermocouple is used to monitor the temperature of the nucleic acid amplification reagent and upload the data to the microprocessor.

[0017] The microprocessor is used to control the frequency and power of the microwave signal based on the temperature of the nucleic acid amplification reagent.

[0018] Furthermore, it also includes a microwave amplifier disposed between the microwave generator and the temperature heating module;

[0019] The microwave amplifier is used to amplify the microwave signal.

[0020] Furthermore, a heat sink is provided above the microwave amplifier.

[0021] Furthermore, it also includes an excitation light source connected to the microprocessor;

[0022] The excitation light source is located on one side of the nucleic acid amplification container and is used to irradiate the solution in the nucleic acid amplification container so that the nucleic acid dye in the solution emits fluorescence of a specific wavelength after receiving the excitation light.

[0023] Furthermore, it also includes a camera connected to the microprocessor;

[0024] The camera is positioned directly above the nucleic acid amplification container and is used to capture fluorescence images and transmit them to the microprocessor.

[0025] Furthermore, the microprocessor is also used to separate the fluorescence image generated by nucleic acid amplification into images of three different channels, and calculate the gray value to obtain the fluorescence intensity.

[0026] Furthermore, the planar microwave resonator is disposed at the bottom of the nucleic acid amplification container.

[0027] Furthermore, the nucleic acid amplification container is disposed inside the waveguide container.

[0028] Furthermore, the microwave generator is externally powered;

[0029] The microprocessor is also used to reduce the voltage of the power supply or cut off the power supply when the temperature of the nucleic acid amplification reagent exceeds a set range.

[0030] The second aspect of this invention provides a method for operating an integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in the first aspect, comprising the following steps:

[0031] A microwave generator outputs microwave signals of a specific frequency and power.

[0032] The temperature heating module transmits microwave signals directionally to the nucleic acid amplification reagent in the nucleic acid amplification container, heating the nucleic acid amplification reagent to the required temperature.

[0033] Thermocouples monitor the temperature of nucleic acid amplification reagents and upload the data to the microprocessor;

[0034] The microprocessor controls the frequency and power of the microwave signal based on the temperature of the nucleic acid amplification reagent.

[0035] Compared with the prior art, the beneficial effects of the present invention are:

[0036] 1. The fully automated nucleic acid amplification system with integrated microwave temperature control described in this invention has a high energy conversion rate: different materials have different absorption effects on microwaves, metals do not absorb them, and polar molecules have a strong absorption rate, which leads to the selective heating of the solution by microwaves. Microwave energy is directly converted into heat energy, resulting in a high energy conversion rate and low loss.

[0037] 2. The integrated microwave temperature-controlled fully automated nucleic acid amplification system described in this invention is miniaturized and portable: the use of a microwave resonator or waveguide allows for targeted heating of liquids in a small space, reducing the volume of heated samples; at the same time, the use of a miniature camera and excitation light source reduces the system size, which is conducive to portable applications.

[0038] 3. The fully automated nucleic acid amplification system with integrated microwave temperature control described in this invention has low cost: the planar resonator and waveguide metal cavity are simple to manufacture, avoiding the use of large instruments and shortening the operation steps; at the same time, the small-volume, low-frequency microwave generator not only effectively reduces costs, but also improves the power output and frequency accuracy of the device.

[0039] 4. The fully automated nucleic acid amplification system with integrated microwave temperature control described in this invention has high temperature control accuracy and short detection time: the temperature of multiple system modules is controlled by microprocessor, and real-time monitoring and feedback adjustment are performed, which improves the temperature control accuracy, increases the temperature regulation rate, and shortens the time required for nucleic acid amplification.

[0040] 5. The fully automated nucleic acid amplification system with integrated microwave temperature control described in this invention uses microwave excitation to generate heat through friction between water molecules, which does not produce a temperature gradient and solves the problem of uneven temperature gradient caused by heat conduction in traditional heating methods. Attached Figure Description

[0041] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments and descriptions of the invention are used to explain the invention and do not constitute an improper limitation of the invention.

[0042] Figure 1 This is a schematic diagram of the principle of the integrated microwave temperature-controlled fully automated nucleic acid amplification system according to Embodiment 1 of the present invention;

[0043] Figure 2 This is a schematic diagram of a planar microwave resonator according to Embodiment 1 of the present invention;

[0044] Figure 3This is a schematic diagram of the fully automated nucleic acid amplification system based on a planar resonator according to Embodiment 1 of the present invention;

[0045] Figure 4 This is a schematic diagram of a waveguide-based fully automated nucleic acid amplification system according to Embodiment 1 of the present invention. Detailed Implementation

[0046] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0047] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0048] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0049] In this invention, terms such as "upper," "lower," "left," "right," "front," "back," "vertical," "horizontal," "side," and "bottom" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. These terms are used only to facilitate the description of the structural relationships of the various components or elements of this invention and do not specifically refer to any component or element in this invention. They should not be construed as limiting the invention.

[0050] In this invention, terms such as "fixed connection," "connected," and "linked" should be interpreted broadly, indicating a fixed connection, an integral connection, or a detachable connection; a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can determine the specific meaning of these terms in this invention based on the specific circumstances, and they should not be construed as limitations on the invention.

[0051] Where there is no conflict, the embodiments and features in the embodiments of the present invention can be combined with each other.

[0052] Example 1

[0053] Embodiment 1 of the present invention provides an integrated microwave temperature-controlled fully automated nucleic acid amplification system.

[0054] Microwaves are electromagnetic waves with frequencies between 0.3 and 300 GHz. Under the influence of a microwave electromagnetic field, the distribution of electron density (or its localized state) changes rapidly, inducing changes in molecular polarization and leading to orientation polarization. Subsequently, water molecules vibrate and rotate rapidly under the influence of the electromagnetic field, generating heat through intermolecular friction. Microwave heating utilizes microwave signals to directly heat nucleic acid solutions, achieving high transmission efficiency (up to 95%) and ensuring the uniformity of the heating curve. Simultaneously, microwaves have penetrating power, enabling simultaneous external and internal heating, rapidly converting microwave energy into internal energy. To reduce system heating time, microwave energy is transferred to the liquid, achieving rapid heating. Microwave heating offers significant advantages, including highly selective absorption, efficient energy conversion, non-contact transfer, and uniform heating.

[0055] Therefore, this embodiment designs an integrated microwave-controlled nucleic acid amplification system by combining microwave temperature control technology and a microwave resonator. The device generates a specific frequency microwave signal through a series connection of a radio frequency generator and a signal amplifier. This signal is then directionally transmitted to the nucleic acid amplification reagent using a planar microwave resonator and waveguide, achieving a rapid temperature rise. Simultaneously, a microprocessor (such as a microcontroller, Raspberry Pi, mobile phone, or computer) monitors the temperature of the nucleic acid amplification solution in real time and precisely controls the sample temperature by adjusting the microwave transmitter power or frequency based on feedback, thus shortening the time required for nucleic acid amplification. Finally, a microprocessor controls a miniature camera to perform fluorescence visualization and quantitative detection of the nucleic acid amplification reaction, involving image capture, processing, and analysis. The detection results are acquired by a microprocessor integrating WiFi and Bluetooth modules and directly transmitted to the cloud for data storage.

[0056] like Figure 1 As shown in the figure, this embodiment provides an integrated microwave temperature-controlled fully automated nucleic acid amplification system, including: a microwave generation module, a signal amplification module, a temperature heating-nucleic acid amplification module (including a temperature heating module and a nucleic acid amplification module), a temperature control module, and an image processing and analysis module (hereinafter referred to as the image processing module).

[0057] The microwave generator module uses the Max 2870 signal generator chip (i.e., microwave generator) 8 to output microwave signals of specific frequency and power. The output frequency of the microwave generator module is 23.5MHz-6000MHz, and the output power is -4dBm-5dBm. It is controlled in real time through serial communication.

[0058] The signal amplification module uses microwave amplifier 2 to amplify the microwave signal. The amplified microwave signal is then transmitted to the planar microwave resonator 10 or the waveguide cavity of the waveguide container 4 via the SMA connection line.

[0059] The nucleic acid amplification module uses a nucleic acid amplification container 6.

[0060] The temperature heating module uses the waveguide component of the waveguide container 4 or the planar microwave resonator 10 to heat the solution inside the nucleic acid amplification container 6.

[0061] The temperature control module uses thermocouples to monitor the temperature of the solution in the nucleic acid amplification container 6 in real time and feeds it back to the microprocessor 9. The microprocessor 9 also uses feedback control to control the microwave generator 8, controlling the frequency and power of the microwave. The microprocessor 9 also uses feedback control to control the signal amplification module, thereby controlling the power of the signal amplifier by controlling the voltage input of the signal amplification module.

[0062] The image processing and analysis module mainly includes an excitation light source 7 and a miniature camera 5. The excitation light source 7 illuminates the nucleic acid amplification container 6, causing the nucleic acid dye in the solution to emit fluorescence of a specific wavelength after receiving the excitation light. The miniature camera 5 captures the fluorescence image and transmits it to the microprocessor 9, which processes the fluorescence image generated by nucleic acid amplification within the nucleic acid amplification container 6 into a digital signal.

[0063] The microprocessor 9 is connected to the microwave generator 8, the microwave amplifier 2, the excitation light source 7, the miniature camera 5, and the thermocouple.

[0064] like Figure 3 and Figure 4 As shown, microwave amplifier 2 is connected to planar microwave resonator 10 or waveguide container 4 via SMA connecting line 3. A heat sink 1 is provided on top of microwave amplifier 2.

[0065] The nucleic acid amplification container 6 is positioned above the planar microwave resonator 10 or inside the waveguide container 4. The miniature camera 5 is positioned directly above the nucleic acid amplification container 6. The excitation light source 7 is positioned on one side of the nucleic acid amplification container 6. The thermocouple is embedded in the inner wall of the nucleic acid amplification container 6.

[0066] Among them, waveguide container 4 is a metal cavity. The metal will totally reflect the microwave signal, and the microwave will enter the nucleic acid amplification region through the metal cavity.

[0067] like Figure 2 As shown, the planar microwave resonator 10 includes a substrate 11 and a metal inner ring 12 and a metal outer ring 13 disposed on the substrate 11. Both the metal inner ring 12 and the metal outer ring 13 are open-ended circular rings, and the metal inner ring 12 is disposed inside the metal outer ring 13. The opening positions of the metal inner ring 12 and the metal outer ring 13 are not the same. The opening position of the metal inner ring 12 is the microwave heating area 15, and the two ends of the opening of the metal outer ring 13 are SMA connection ports 14.

[0068] The substrate 11 can be made of glass, epoxy resin board, etc., with no material restrictions. Any solid material can be used as the substrate.

[0069] The inner metal ring 12 and the outer metal ring 13 are both made of metal materials such as copper foil and copper mold, and have a certain thickness. Microwave signals are transmitted through the metal materials.

[0070] The implementation method of an integrated microwave temperature-controlled fully automated nucleic acid amplification system provided in this embodiment is as follows:

[0071] (1) The microwave generator module uses the Max 2870 microwave generator chip to output microwave signals of specific frequency and power. The microwaves are transmitted to the microwave amplifier through the SMA connection line.

[0072] The frequency of the microwave signal is adjusted according to the structure of the planar microwave resonator or waveguide. Different microwave resonators and waveguide structures have different resonant frequencies, which in turn change the frequency at which water molecules vibrate. The effective frequency used in this embodiment is 1.2-1.8 GHz.

[0073] The microwave amplifier requires an external power supply of 12-24V, which is controlled in real time by a microprocessor. If the temperature is too high, the microprocessor reduces the power supply voltage or cuts off the power; if the temperature is too low, it increases the voltage. Simultaneously, the microprocessor can also control the temperature by adjusting the output frequency and power of the Max2870 signal generator.

[0074] (2) The amplified microwave signal is transmitted to a planar microwave resonator or waveguide cavity via an SMA connector, both of which contain the actual container for nucleic acid amplification. The microwaves cause the water molecules in the nucleic acid reagent to vibrate violently, resulting in intense friction between the molecules and a rapid rise in temperature.

[0075] (3) The thermocouple monitors the temperature change in real time and transmits the temperature data to the microprocessor. The microprocessor then emits feedback control signals to control the frequency and power of the microwave, or to control the reduction or cut-off voltage of the external power supply of the microwave amplifier, thereby regulating the temperature.

[0076] (4) The reagents in nucleic acid amplification container 6 are the reaction reagents required for nucleic acid amplification, including dNTPs, DNA polymerase, trehalose, Tris HCl, nucleic acid dye, and Mg2+. 2+ Reaction buffers, etc.

[0077] (5) The image processing module mainly includes an excitation light source and a miniature camera. The microprocessor 9 controls the excitation light source to irradiate the nucleic acid amplification container. After receiving the excitation light, the nucleic acid dye in the solution emits fluorescence at a specific wavelength. The microprocessor 9 controls the camera to capture fluorescence images or videos in real time and transmits the information to the microprocessor 9. The microprocessor 9 separates the fluorescence image (color) into images of three different channels (red channel, blue channel, and green channel) and calculates the gray value to achieve quantitative detection of fluorescence intensity. The fluorescence intensity and the gray value of the specific channel have a linear relationship.

[0078] (6) The microprocessor uploads data to the cloud storage via its own output ports (USB interface, Bluetooth module, WiFi module) or displays the fluorescence intensity of real-time nucleic acid reagent amplification on the screen.

[0079] In this embodiment, the substrate for fabricating the planar microwave resonator includes, but is not limited to, glass, PCB board, and epoxy resin board.

[0080] In this embodiment, the excitation light source includes, but is not limited to, a single-wavelength laser and a monochrome LED.

[0081] In this embodiment, the microprocessor block includes, but is not limited to, a USB interface, a Bluetooth module, and a WiFi module.

[0082] This embodiment provides an integrated microwave temperature-controlled fully automated nucleic acid amplification system that combines microwave temperature control technology with nucleic acid amplification technology. It generates a specific frequency microwave signal through a series connection of a radio frequency generator and a signal amplifier, and then uses a planar microwave resonator, waveguide, or other means to directionally transmit the microwave signal to the nucleic acid amplification reagent to achieve a rapid temperature rise.

[0083] The fully automated nucleic acid amplification system with integrated microwave temperature control provided in this embodiment has the following advantages:

[0084] (1) High energy conversion rate: Different materials have different absorption effects on microwaves. Metals do not absorb microwaves, polar molecules have a strong absorption rate, nucleic acid solutions are aqueous solutions, and water is a polar molecule that almost completely absorbs microwave signals, resulting in microwaves selectively heating the solution. Microwave energy is directly converted into heat energy, with a high energy conversion rate and low loss.

[0085] (2) Miniaturization and portability: The use of microwave resonators or waveguides allows for targeted heating of liquids in a small space, reducing the volume of heated samples; at the same time, the use of miniature cameras and excitation light sources reduces the system size, which is helpful for portable applications.

[0086] (3) Low cost: Planar resonators and waveguide metal cavities are easy to manufacture, avoid the use of large instruments and shorten the operation steps; at the same time, small volume, low frequency microwave generators not only effectively reduce costs, but also improve the power output and frequency accuracy of the device.

[0087] (4) High temperature control accuracy and short detection time: The temperature of multiple system modules is controlled by microprocessor, and real-time monitoring and feedback adjustment are achieved, which improves the temperature control accuracy, increases the temperature regulation rate, and shortens the time required for nucleic acid amplification.

[0088] (5) The use of microwave to excite water molecules to generate heat through friction does not produce a temperature gradient, thus solving the problem of uneven temperature gradient caused by heat conduction in traditional heating methods.

[0089] Example 2

[0090] Embodiment 2 of the present invention provides a working method for an integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in Embodiment 1, comprising the following steps:

[0091] (1) The microwave generator module uses the Max 2870 microwave generator chip to output microwave signals of specific frequency and power. The microwaves are transmitted to the microwave amplifier through the SMA connection line.

[0092] The frequency of the microwave signal is adjusted according to the structure of the planar microwave resonator or waveguide.

[0093] The microwave generator requires an external 12-24V power supply, which is controlled in real-time by a microprocessor. If the temperature is too high, the microprocessor reduces or cuts off the voltage; if the temperature is too low, it increases the voltage. Simultaneously, the microprocessor can also control the temperature by adjusting the output frequency and power of the Max2870 signal generator.

[0094] (2) The amplified microwave signal is transmitted to a planar microwave resonator or waveguide cavity via an SMA connector, both of which contain the actual container for nucleic acid amplification. The microwaves cause the water molecules in the nucleic acid reagent to vibrate violently, resulting in intense friction between the molecules and a rapid rise in temperature.

[0095] (3) The thermocouple monitors the temperature change in real time and transmits the temperature data to the microprocessor. The microprocessor then emits feedback control signals to control the frequency and power of the microwave, or to control the reduction or cut-off voltage of the external power supply of the microwave amplifier, thereby regulating the temperature.

[0096] (4) The reagents in nucleic acid amplification container 6 are the reaction reagents required for nucleic acid amplification, including dNTPs, DNA polymerase, trehalose, Tris HCl, nucleic acid dye, and Mg2+. 2+ Reaction buffers, etc.

[0097] (5) The image processing module mainly includes an excitation light source and a miniature camera. The microprocessor 9 controls the excitation light source to irradiate the nucleic acid amplification container. After receiving the excitation light, the nucleic acid dye in the solution emits fluorescence at a specific wavelength. The microprocessor 9 controls the camera to capture fluorescence images or videos in real time and transmits the information to the microprocessor 9. The microprocessor 9 separates the fluorescence image (color) into images of three different channels (red channel, blue channel, and green channel) and calculates the grayscale value to achieve quantitative detection of fluorescence intensity.

[0098] (6) The microprocessor uploads data to the cloud storage via its own output ports (USB interface, Bluetooth module, WiFi module) or displays the fluorescence intensity of real-time nucleic acid reagent amplification on the screen.

[0099] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. An integrated microwave temperature-controlled fully automated nucleic acid amplification system, characterized in that: It includes a microprocessor, a temperature heating module, a nucleic acid amplification container, and a microwave generator and a thermocouple connected to the microprocessor. The microwave generator is connected to the temperature heating module, and the temperature heating module adopts a planar microwave resonator or a waveguide container. The microwave generator is used to output microwave signals of a specific frequency and power; the output frequency of the microwave generator is 23.5MHz-6000MHz, and the output power is -4dBm-5dBm. The temperature heating module is used to transmit microwave signals in a directional manner to the nucleic acid amplification reagent in the nucleic acid amplification container, thereby heating the nucleic acid amplification reagent to a certain temperature. The thermocouple is used to monitor the temperature of the nucleic acid amplification reagent and upload the data to the microprocessor. The microprocessor is used to control the frequency and power of the microwave signal according to the temperature of the nucleic acid amplification reagent; The planar microwave resonator is disposed at the bottom of the nucleic acid amplification container; A planar microwave resonator directs microwave signals into nucleic acid amplification reagents to achieve a rapid temperature rise. The planar microwave resonator includes a substrate and an inner metal ring and an outer metal ring disposed on the substrate. Both the inner and outer metal rings are open circular rings, and the inner metal ring is disposed inside the outer metal ring. The opening positions of the inner and outer metal rings are not the same. The opening position of the inner metal ring is the microwave heating area, and the two ends of the opening of the outer metal ring are SMA connection ports.

2. The fully automated nucleic acid amplification system with integrated microwave temperature control as described in claim 1, characterized in that: It also includes a microwave amplifier disposed between the microwave generator and the temperature heating module; The microwave amplifier is used to amplify the microwave signal.

3. The integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in claim 2, characterized in that: A heat sink is provided on top of the microwave amplifier.

4. The integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in claim 1, characterized in that: It also includes an excitation light source connected to the microprocessor; The excitation light source is located on one side of the nucleic acid amplification container and is used to irradiate the solution in the nucleic acid amplification container so that the nucleic acid dye in the solution emits fluorescence of a specific wavelength after receiving the excitation light.

5. The integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in claim 1, characterized in that: It also includes a camera connected to the microprocessor; The camera is positioned directly above the nucleic acid amplification container and is used to capture fluorescence images and transmit them to the microprocessor.

6. The fully automated nucleic acid amplification system with integrated microwave temperature control as described in claim 1, characterized in that: The microprocessor is also used to separate the fluorescence image generated by nucleic acid amplification into three different channels and calculate the gray value to obtain the fluorescence intensity.

7. The fully automated nucleic acid amplification system with integrated microwave temperature control as described in claim 1, characterized in that: The nucleic acid amplification container is disposed inside the waveguide container.

8. The fully automated nucleic acid amplification system with integrated microwave temperature control as described in claim 1, characterized in that: The microwave generator is connected to an external power supply; The microprocessor is also used to reduce the voltage of the power supply or cut off the power supply when the temperature of the nucleic acid amplification reagent exceeds a set range.

9. The operating method of an integrated microwave temperature-controlled fully automated nucleic acid amplification system as described in any one of claims 1-8, characterized in that: Includes the following steps: A microwave generator outputs microwave signals of a specific frequency and power. The temperature heating module transmits microwave signals directionally to the nucleic acid amplification reagent in the nucleic acid amplification container, heating the nucleic acid amplification reagent to the required temperature. Thermocouples monitor the temperature of nucleic acid amplification reagents and upload the data to the microprocessor; The microprocessor controls the frequency and power of the microwave signal based on the temperature of the nucleic acid amplification reagent.

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