Analysis device and detection method based on gene detection

Abstract

The invention discloses an analysis device based on gene detection and a detection method, and relates to the field of biological detection. The invention relates to a fluorescent quantitative PCR analysis device, and aims to solve the problems that the existing fluorescent quantitative PCR device is low in heating efficiency, a reaction system is difficult to quickly reach a target temperature, the detection period is prolonged, the consistency of PCR reaction is influenced and the like, the analysis device comprises a reaction chamber module, a reaction chamber rotary table module, a single chip microcomputer module and the like, and regular room-temperature air is conveyed through a top cover fan; a heating wire in a heating middle cylinder is heated into hot air, the hot air is uniformly dispersed by a bottom fan, all reaction holes are uniformly heated, the hot air passes through the reaction holes and is discharged out of a reaction chamber from a hollow position of an elastic gland, a top cover is connected with the bottom of a machine body through a hydraulic rod, and fluorescence data of all the reaction holes are collected through a fluorescence detection module; the temperature of the reaction chamber is controlled through the single-chip microcomputer module, the rotating speed and the position of the servo motor are adjusted, the time sequence of a fluorescence collection optical system is matched, and the device is suitable for the fields of material protection under high-temperature or variable-temperature working conditions.

Description

Technical Field

[0001] The present invention relates to the field of biological detection technology, and in particular to an analysis device and a detection method based on gene detection. Background Art

[0002] As a core technology in molecular biology, polymerase chain reaction (PCR) plays a vital role in medical diagnosis, disease prevention, and biological research. Fluorescence quantitative PCR technology further enables real-time monitoring of the PCR process and quantitative analysis of nucleic acid templates, providing a powerful tool for precise detection.

[0003] However, the mainstream fluorescent quantitative PCR devices currently on the market have obvious defects. In terms of heating methods, although semiconductor heating has a certain degree of temperature control accuracy, its heating efficiency is low, making it difficult to quickly make the reaction system reach the target temperature, which prolongs the detection cycle. In addition, its heating is uneven, resulting in temperature differences in different reaction areas, affecting the consistency of the PCR reaction and reducing the accuracy of the test results. Although water bath heating can create a relatively uniform temperature environment, the equipment is bulky and takes up a lot of space, which is not conducive to the efficient use of laboratory space. It also has high energy consumption and high operating costs. At the same time, water baths are prone to breeding bacteria and require frequent cleaning and maintenance, which not only increases the operational burden, but may also introduce contamination, affecting the reliability of the test results.

[0004] The reaction well positioning accuracy of existing devices is insufficient, and deviations are prone to occur during movement, making it impossible to accurately locate each working area, affecting the reaction process. The overall operating stability of the device is poor, and long-term operation is prone to problems such as vibration and loose components, which reduces the service life and reliability of the equipment. The coordination between the various functional modules is poor, and efficient coordination cannot be achieved, resulting in detection efficiency that is difficult to meet the growing detection needs. Therefore, the development of an analysis device based on genetic testing to solve the above-mentioned technical problems has important practical significance and application value. Summary of the Invention

[0005] The present invention aims to solve the problems of low heating efficiency of existing fluorescent quantitative PCR devices, which makes it difficult to quickly bring the reaction system to the target temperature, prolonging the detection cycle, and uneven heating, resulting in temperature differences in different reaction areas, affecting the consistency of PCR reactions and reducing the accuracy of detection results. The equipment is bulky and occupies a lot of space, which is not conducive to the efficient use of laboratory space, and has high energy consumption and high operating costs.

[0006] To solve the above technical problems, the present invention is achieved through the following technical solutions: Solution 1: The present invention proposes an analysis device based on genetic testing, which includes a synchronous belt reaction chamber turntable module, a reaction chamber module, a servo motor module, a top cover module, an external support module, a power module, a top fan module, a fluorescence detection module, a heating middle cylinder, a single-chip microcomputer module, and a body base; The top fan module includes a fan bracket, a top fan body, a spoiler module and a wind speed adjustment module; The top cover module includes a top cover body, a sealing arc strip, a heating element and an elastic pressure cover; The computer sends instructions to the single-chip microcomputer module, which controls the temperature inside the reaction chamber module. The top fan body delivers regular room temperature air, which is heated to the required hot air through the heating wire in the middle heating cylinder of the reaction chamber module. The bottom fan evenly disperses the hot air and evenly heats all reaction holes. The hot air passes through the reaction holes and is discharged from the hollow position of the elastic pressure cover to the reaction chamber, and is discharged from the exhaust channel between the top cover body and the fuselage. The top cover body is connected to the bottom of the fuselage through a hydraulic rod. During the reaction, the servo motor module drives the synchronous belt reaction chamber turntable module. The fluorescence detection module is a four-channel multi-wavelength simultaneous detection optical module. The bottom four-channel multi-wavelength simultaneous detection optical module collects fluorescence data of each reaction hole. The single-chip microcomputer module receives and processes the collected fluorescence data of each reaction hole, and outputs and displays the processed fluorescence data through the host computer.

[0007] Furthermore, a preferred embodiment is provided, wherein the fan bracket is fixedly installed above the top cover module, and the wind speed regulating device is connected to the top fan body.

[0008] Furthermore, a preferred embodiment is provided, wherein a hydraulic rod buffer module is also provided between the top cover body and the fuselage base, and the hydraulic rod buffer module includes a hydraulic rod, a fixing seat and a connecting module; the fixing seat is fixedly installed on the top cover body and the fuselage respectively, and the two ends of the hydraulic rod are respectively hinged to the bottom fixing seat and the connecting module.

[0009] Furthermore, a preferred embodiment is provided, wherein a top cover locking module is further provided between the top cover body and the fuselage base, the top cover locking module comprising a locking shaft, an infrared radiation sensor, a linkage rod and a lock assembly; the lock assembly comprising a lock tongue and a lock seat; The locking shaft is fixed to the inner side of the top cover body and is connected to the top cover body through the end hexagonal nut. One end of the linkage rod is connected to the lock seat, and the other end is inserted into the middle of the infrared radiation sensor to block infrared transmission when the instrument is closed; the lock seat is fixed to the fuselage, and the lock tongue is extended and retracted by the locking shaft and engages or separates with the lock seat; by operating the servo to rotate the lock seat, the lock seat is driven to engage with the lock tongue, thereby achieving tight locking of the top cover body and the fuselage.

[0010] Furthermore, a preferred embodiment is provided, in which the synchronous belt reaction chamber turntable module includes a driving wheel, a synchronous transmission belt, a rotating table, and a tensioning wheel. The driving wheel is connected to the output shaft of the servo motor module through a high-precision coupling, and the tensioning wheel is installed on the guide rail of the external support module. The engagement between the driving wheel and the synchronous transmission belt drives the synchronous transmission belt to perform a circular motion, passing through the detection holes of the fluorescence optical detection module at the bottom of the reaction chamber in sequence.

[0011] Furthermore, a preferred embodiment is provided, wherein the servo motor module includes a servo motor, a driver and an encoder; The encoder is installed on the output shaft of the servo motor to detect the rotation position and speed of the servo motor in real time; The signal detected by the encoder is fed back to the driver in the form of pulses, providing the driver with the actual operating status information of the servo motor.

[0012] Furthermore, a preferred embodiment is provided, in which the sealing arc strip is installed on the edge of the top cover body. When the top cover body is matched with the reaction chamber module, the elastic pressure cover is used to fill the gap between the top cover body and the reaction chamber module to form a sealing structure. The heating element is realized by a surrounding resistance wire and is evenly laid inside the heating cylinder.

[0013] Furthermore, a preferred embodiment is provided, wherein the external support module includes a middle support plate, support columns and a base plate, and the base plate, the middle support plate and support columns are assembled into a frame structure by bolt connection and corner piece fixation, and the support columns and the base plate are provided with at least one mounting hole and a guide rail, the mounting hole is used to fix the reaction chamber module and synchronously drive the reaction chamber turntable module, the servo motor module and the top cover module; the guide rail is used to provide guidance for the tensioning wheel.

[0014] Furthermore, a preferred embodiment is provided, wherein the power supply module includes a transformer, a rectifier circuit, a filter circuit and a voltage stabilizing circuit, and is used to provide electrical energy to the single-chip microcomputer module, the servo motor module and the heating element.

[0015] Solution 2: A detection method implemented by the gene detection-based analysis device described in any one of Solution 1, the detection method comprising the following steps: Step 1: First, pre-treat the nucleic acid sample to be tested, use a pipette to transfer a certain amount of reaction solution to the microfluidic chip, and fix the PCR reaction chamber through the reaction chamber module and the synchronous belt reaction chamber turntable module so that the central axis of the reaction chamber module is perpendicular to the running direction of the synchronous belt; Step 2: Start the single-chip microcomputer module, first calibrate the temperature sensor, and correct the temperature sensor error using a standard temperature source; then reset the servo motor module, and determine the initial positions of the reaction chamber module and the synchronous belt reaction chamber turntable module using the encoder feedback signal; the user enters the PCR reaction program parameters through the host computer software, including the denaturation temperature, annealing temperature, extension temperature, hold time for each stage, and number of cycles, and also sets the wind speed of the top fan module during the heating stage and the auxiliary wind speed during the cooling stage; Step 3. The single-chip computer module sends a positioning instruction to the servo motor module. The servo motor module drives the synchronous belt through the active wheel to align the microfluidic chip reaction pool loaded with PCR reaction solution with the fluorescence detection hole. After positioning is completed, the lock seat of the top cover locking module is operated to rotate, driving the lock tongue of the lock assembly to engage with the lock seat on the body. The linkage rod rotates with the lock seat to block the infrared radiating tube. At the same time, the hydraulic rod buffer module is compressed and contracted to ensure that the elastic pressure cover top cover is tightly fitted with the top surface of the reaction chamber module, providing conditions for uniform heating. Step 4: The single-chip microcomputer module controls the top cover heating element and the fan module to start. The heating power is dynamically adjusted according to the volume of the reaction chamber module and the heating rate requirement. At the same time, the top fan module runs at the set wind speed to drive the air in the reaction chamber module to form a convection circulation; Step 5: Real-time detection of fluorescence signals. After the extension phase of each PCR cycle is completed, the fluorescence detection module is activated. The excitation light emitted by the excitation light source is transmitted to the common end through the combined end of the bifurcated optical fiber. The common end is incident on the detection area, stimulating the fluorescent probe in the reaction solution to produce fluorescence. The emitted fluorescence signal is collected by the same common end and transmitted to the fluorescence detector through the splitting end of the bifurcated optical fiber. The fluorescence detector converts the optical signal into an electrical signal, which is then processed in real time by the single-chip microcomputer module after A / D conversion. Step 6. Repeat steps 3 to 5 until the preset number of cycles is reached. Before each cycle starts, the microcontroller module automatically calibrates the servo motor position to ensure that the reaction well position is accurately reset; at the same time, the heating element power and fan speed are adaptively adjusted according to the temperature response data of the previous cycle; when the number of cycles reaches the set value, the system automatically enters the termination program; the result analysis and data output microcontroller module processes the fluorescence data of all cycles; the test results are transmitted to the host computer software through the USB, RS-485, and WiFi communication interfaces to generate a test report containing the Ct value, concentration, amplification curve, and melting curve.

[0016] The present invention is beneficial in that: The genetic testing-based analysis device and detection method described in the present invention utilizes hot air heating, combined with the heating element of the top cover module, to achieve uniform temperature distribution within the reaction chamber, reduce temperature gradients, and improve the consistency and accuracy of PCR reactions. Precise control of the reaction chamber's synchronous belt rotation module and servo motor module enables simultaneous detection of multiple reaction cells, enabling rapid and continuous testing of multiple samples, thereby improving detection efficiency. Automated control of the entire device is achieved through a single-chip microcomputer module; users only need to set the PCR reaction program and related parameters to complete the detection process, resulting in simple operation and ease of use.

[0017] The present invention is also applicable to practical engineering application fields such as material protection under high temperature or variable temperature conditions, cavitation effect regulation, and rapid detection and analysis of multiple genes. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a cross-sectional view of the overall structure of an analysis device based on genetic testing as described in embodiment 1.

[0019] Figure 2 This is a side view of a transmission module of a gene detection-based analysis device according to embodiment 1.

[0020] Figure 3 A front view of a locking module of a gene detection-based analysis device according to embodiment 1 Figure 4 A cross-sectional view of a locking module of a gene detection-based analysis device according to embodiment 1 Figure 5 This is a flow chart of a detection method implemented by a gene detection-based analysis device as described in Embodiment 3.

[0021] Among them, there are fan bracket 1, top fan body 2, heating middle cylinder 3, top cover body 4, elastic pressure cover 5, reaction chamber module 6, middle support plate 7, support column 8, foot 9, bottom plate 10, single chip computer module 11, servo motor module 12, heat dissipation module 13, power module 14, hydraulic rod 15, connection module 16, top support plate 17, driving wheel 18, synchronous transmission belt 19, rotating table 20, tensioning wheel 21, locking shaft 22, infrared radiation sensor 23, linkage rod 24, and locking assembly 25. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solutions and advantages of the implementation methods of this application clearer, the technical solutions in the implementation methods of this application will be clearly and completely described below in combination with the drawings in the implementation methods of this application. Obviously, the described implementation methods are only part of the implementation methods of this application, not all of the implementation methods.

[0023] Embodiment 1. This embodiment proposes an analysis device based on genetic testing, which includes a fan bracket 1, a top fan body 2, a heating middle cylinder 3, a top cover body 4, an elastic pressure cover 5, a reaction chamber module 6, a middle support plate 7, a support column 8, a base 9, a base plate 10, a single-chip computer module 11, a servo motor module 12, a heat dissipation module 13, a power module 14, a hydraulic rod 15, a connection module 16, a top support plate 17, a driving wheel 18, a synchronous transmission belt 19, a rotating table 20, a tensioning wheel 21, a locking shaft 22, an infrared radiation sensor 23, a linkage rod 24, and a locking assembly 25.

[0024] The external computer sends instructions to the single-chip computer module 11, and the single-chip computer module 11 runs the internal program according to the instructions, controls the temperature inside the reaction chamber module 6, controls the servo motor module 12 to drive the rotating table 20, realizes the rotation of the microfluidic chip in the reaction chamber module 6, and enables the optical system to detect the reaction pools at different positions. The collected signals are received and processed by the single-chip computer, and communicate with the host computer to output and realize the signals. The reaction chamber module 6, the rotating table 20, the single-chip computer module 11, the servo motor module 12, the top cover body 4, the middle support plate 7, the top support plate 17, and the power supply module 14 are all inside the top cover body 4 and the external support module.

[0025] The fan module includes a fan bracket 1 and a top fan body 2. The top fan body 2 adopts a centrifugal fan, which has the characteristics of large air volume and low noise, and can quickly generate airflow. The fan bracket 1 is fixedly installed above the top cover body 4, and is used to support the top fan body 2 to ensure the stability of the fan operation. The wind speed adjustment device is connected to the top fan body 2, and can adjust the wind speed of the fan through the control instructions of the single-chip module 11 according to the different stages of the PCR reaction. In the heating stage of the PCR reaction, the single-chip module 11 controls the start-up of the top fan module, and the top fan body 2 generates airflow, accelerates the air flow in the reaction chamber, and enables heat to be transferred to various parts of the reaction chamber more quickly and evenly, thereby improving the heating efficiency. In the cooling stage, the fan can assist the cooling system by adjusting the wind speed, accelerate the cooling rate of the reaction chamber, and shorten the cooling time. By precisely controlling the wind speed and start and stop of the fan, the temperature change rate of the reaction chamber can be effectively regulated, and the PCR reaction process can be further optimized. A hydraulic rod buffer module is also included between the top cover body 4 and the base plate 10, and the hydraulic rod buffer module includes a hydraulic rod 15 and a connecting module 16; they are fixedly installed on the top cover body 4 and the base plate 10 respectively by means of a fixing seat, and the two ends of the hydraulic rod 15 are hinged to the base plate 10 and the connecting module 16 respectively. During the operation of the device, the hydraulic rod buffer module can buffer the impact force generated when the top cover is opened and closed, reduce the impact of vibration on the internal components of the device, and at the same time ensure that it is tightly fitted with the reaction chamber module 6 when the top cover is closed.

[0026] like Figure 2As shown, the synchronous belt reaction chamber turntable module consists of a driving wheel 18, a rotating table 20, a tensioning wheel 21 and a synchronous transmission belt 19. The synchronous belt is made of high-strength polyurethane material. This material gives the synchronous belt excellent wear resistance, which can withstand long-term high-frequency use without being easily damaged; its transmission efficiency is high, and it can efficiently transmit the power of the servo motor; the noise during operation is low, ensuring a quiet working environment for the device. It can work closely with the PCR reaction chamber to ensure that the reaction chamber remains stable during the operation of the device. The driving wheel 18, the rotating table 20 and the tensioning wheel 21 are all made of aluminum alloy, and the surface is anodized. Anodizing not only improves the wear resistance of the wheel, but also enhances its corrosion resistance and extends its service life. The driving wheel 18 is connected to the output shaft of the servo motor module 12 through a high-precision coupling to ensure the accuracy and stability of power transmission. The tensioning wheel 21 is mounted on a guide rail pre-set by the external support module. By adjusting the position of the tensioning wheel 21 on the guide rail, the tension of the synchronous transmission belt 19 can be precisely adjusted, so that the synchronous belt always maintains the optimal transmission state and avoids slipping or loosening, thereby ensuring the stable operation of the synchronous transmission belt 19 driven by the reaction chamber module 6. After the servo motor module 12 is started, it drives the driving wheel 18 to start rotating. The driving wheel 18 drives the synchronous transmission belt 19 to perform a circular motion by virtue of the engagement between the driving wheel 18 and the synchronous transmission belt 19. The rotating table 20 mounted on the synchronous transmission belt 19 rotates with the synchronous transmission belt 19, allowing the microfluidic chip in the PCR reaction chamber to pass through the detection hole of the fluorescence detection module at the bottom of the reaction chamber in sequence. During this process, the single-chip computer module 11 adjusts the speed and rotation angle of the servo motor module 12 by precisely controlling the servo motor module 12. According to the requirements of the PCR reaction program, the residence position and residence time of the reaction chamber are accurately controlled.

[0027] like Figure 3 As shown, a top cover locking module is also provided between the top cover body 4 and the fuselage, and the top cover locking module includes a locking shaft 22, an infrared radiation sensor 23, a linkage rod 24 and a lock assembly 25; the locking shaft 22 is fixed to the inner side of the top cover and is connected to the top cover body 4 through a hexagonal nut at the end, one end of the linkage rod 24 is connected to the lock seat, and the other end is inserted into the middle of the infrared radiation sensor 23 to block infrared transmission when the instrument is closed; the lock assembly 25 includes a lock tongue and a lock seat, the lock seat is fixed to the fuselage, and the lock tongue can be engaged or separated with the lock seat under the drive of the locking shaft 22; as shown Figure 4 As shown, by operating the servo to rotate the lock seat, the lock seat is driven to engage with the lock tongue, so that the top cover and the fuselage are tightly locked, ensuring that the top cover body 4 of the device is stable during the PCR reaction and preventing heat loss and external contamination. When unlocking, the lock seat can be operated in reverse to make the lock tongue detach from the lock seat.

[0028] The fluorescence detection module is connected to the four light-through holes at the bottom of the shell of the reaction chamber module 6 with screws. The fluorescence detection module is a four-channel multi-wavelength simultaneous detection optical system, which mainly uses a special structure of combining light at one end and splitting light at the other end of the bifurcated optical fiber. Its common end undertakes the core function of exciting the sample and receiving the fluorescence signal during the entire detection process. Specifically, the combining end of the bifurcated optical fiber is connected to a high-stability excitation light source through a high-precision optical interface to ensure efficient transmission and conversion of the optical signal; and the splitting end is based on the reflection grating inside the device to enable multi-channel fluorescence detection to achieve simultaneous splitting. When working, the excitation light source emits high-intensity excitation light of a specific wavelength, which is transmitted along the combining end of the bifurcated optical fiber and reaches the common end with almost no loss due to the good light guiding performance of the optical fiber. Subsequently, the common end will shoot the excitation light at a precise angle and intensity to the sample in the PCR reaction chamber. The fluorescent groups in the sample are activated by the excitation light, thereby generating fluorescence. The generated fluorescence is also collected through the common end. After preliminary aggregation and organization of the fluorescence signal, the common end leverages the transmission capacity of the bifurcated fiber to stably transmit the fluorescence signal to the reflective grating connected to the splitting end for splitting, ultimately converging at different locations on the infrared radiation sensor 23. During this process, the unique structure of the bifurcated fiber ensures efficient transmission of the excitation light and fluorescence signal, greatly improving the accuracy and sensitivity of fluorescence detection. This enables high-precision, high-efficiency detection and stable transmission of the fluorescence signal during the PCR reaction, providing reliable data support for real-time monitoring and quantitative analysis of the PCR reaction.

[0029] The single-chip microcomputer module 11 utilizes a high-performance microcontroller as its core and integrates multiple key functional modules. The data processing unit possesses powerful computing capabilities, enabling rapid and accurate processing and analysis of signals from various sensors, such as temperature and position sensors. It analyzes sensor data based on pre-set PCR reaction procedures and algorithms, generating corresponding control instructions. The communication interface supports multiple protocols, such as USB and RS-485, facilitating data transmission and communication with external devices. This interface enables remote control of the device, allowing users to operate and configure the device from a remote terminal. It also facilitates data sharing, allowing test results to be transmitted to other devices for further analysis. The A / D conversion module converts analog signals, such as temperature and pressure, into digital signals, enabling them to be recognized and processed by the single-chip microcomputer. The PWM output module specifically controls the speed and rotation angle of the servo motor module 12. By outputting pulse signals of varying frequencies and duty cycles, it achieves precise control of the servo motor, thereby controlling the operation of the reaction chamber's synchronous belt rotation module. During device operation, the single-chip microcomputer module collects key information, such as the reaction chamber's temperature and position, in real time through various connected sensors. This collected information is transmitted to the microcontroller as analog signals, converted to digital signals by the A / D conversion module, and then fed into the data processing unit. The data processing unit performs in-depth analysis and calculations on these digital signals based on the pre-set PCR reaction program and algorithm. Based on the calculation results, it determines the servo motor's speed and rotation angle, as well as the operating parameters of the heating element and cooling system. The corresponding control instructions are then sent to the servo motor module 12 and other related modules via the PWM output module. When the temperature sensor detects that the reaction chamber temperature has not reached the set denaturation temperature, the microcontroller module 11 controls the heating element to increase power. When the reaction chamber module 6 reaches the designated position, the servo motor module 12 is controlled to pause rotation, ensuring that the reaction chamber remains accurately within the fluorescence detection area. Simultaneously, the microcontroller module 11 maintains data exchange with external devices via a communication interface, promptly transmitting real-time data, test results, and device operating status information during the detection process to the user terminal or host computer system for convenient monitoring and management. The servo motor module 12 consists of a servo motor, a driver, and an encoder. The servo motor is a high-precision AC servo motor, characterized by its fast response speed, enabling quick response to control commands; high positioning accuracy, enabling precise control of rotation angle and position; and high output torque, sufficient to drive the reaction chamber synchronous belt rotation module for stable operation. The driver, acting as a bridge between the servo motor and the microcontroller module, receives control commands from the microcontroller module 11 and converts them into drive signals suitable for the servo motor. It precisely adjusts the servo motor's speed, rotation angle, and torque, ensuring that the servo motor operates according to predetermined requirements. The encoder, mounted on the servo motor's output shaft, detects the motor's rotational position and speed in real time. This detected signal is fed back to the driver in the form of pulses, providing information on the servo motor's actual operating status, thus forming a closed-loop control system. After receiving the PWM control signal from the microcontroller module 11, the driver adjusts the servo motor's input voltage and current based on the signal's frequency and duty cycle. By varying the magnitude and direction of the input voltage and current, the servo motor's speed and rotation angle are controlled. As the servo motor rotates, the encoder continuously detects the motor's rotational position and speed and feeds the detection signal back to the driver. The driver compares and analyzes the feedback signal with the control instructions sent by the microcontroller module. If any deviation is detected between the actual operating status and the control instructions, the driver promptly adjusts the servo motor's input signal to quickly align the servo motor's actual operating status with the control instructions. This achieves high-precision and stable control of the reaction chamber's synchronous belt rotation module, ensuring that the PCR reaction chamber can accurately and stably move and stay between the various fluorescence detection wells.

[0030] The top cover body 4 primarily comprises a heating cylinder 3 and an elastic gland 5. The top cover body 4 is made of a high-strength aluminum alloy, a material with high strength that can withstand certain pressures and external forces, ensuring the structural stability of the top cover. A sealing arc strip is installed on the edge of the top cover body. Specially designed and processed, it fits perfectly with the grooves on the body, providing excellent sealing. When the top cover body 4 and the reaction chamber module 6 are mated, the elastic gland 5 tightly fills the gap between them, forming a reliable seal. This prevents heat loss within the reaction chamber and blocks external contaminants from entering the reaction chamber, ensuring a clean PCR reaction environment. The heating cylinder 3 utilizes a surrounding resistance wire, evenly laid inside the top cover heating cylinder, enabling rapid and uniform heating of the reaction chamber top. During the PCR reaction, the top cover module fits tightly with the reaction chamber module 6 via the elastic gland 5, creating a closed space. The single-chip microcomputer module 11 controls the operating state of the heating element according to the requirements of the PCR reaction program. When the reaction reaches the stage where the reaction chamber needs to be heated, the heating element begins operating according to the control instructions and drives the fan to heat the reaction chamber. The external support module serves as the framework for the entire device, playing a key role in supporting and securing the other modules. It primarily consists of a central support plate 7, support columns 8, a base plate 10, and a top support plate 17. The base plate 10 is made of high-strength metal and undergoes precision machining and surface treatment, resulting in exceptional flatness and stability. It can withstand the weight of the entire device, as well as the vibration and impact forces generated during operation, ensuring the device remains stable during operation. The central support plate 7, support columns 8, and top support plate 17 are constructed from aluminum alloy profiles, which offer high strength and light weight. They are assembled into a framework structure using bolt connections and corner fixings. This assembly method not only facilitates installation but also ensures the strength and stability of the framework. The central support plate 7 and top support plate 17 feature multiple precisely machined mounting holes and guide rails for mounting components such as the top cover 4, reaction chamber module 6, and servo motor module 12, ensuring accurate installation. The guide rails provide guidance for adjusting the tensioner 21, ensuring that the synchronous belt drive module does not disengage from the synchronous belt 19.

[0031] The power module 14 consists of a transformer, a rectifier circuit, a filter circuit, and a voltage-stabilizing circuit. The transformer converts the high voltage of the mains input into the appropriate voltage required by each module of the device, meeting the power requirements of each module. The rectifier circuit uses high-efficiency rectifier components to convert the input AC power into DC power, providing a basic power source for subsequent circuits. The filter circuit, composed of components such as capacitors and inductors, primarily removes noise and ripple from the rectified DC power, making the DC power smoother and reducing the impact of power supply fluctuations on the various modules of the device. The voltage-stabilizing circuit stabilizes the DC power according to the actual power requirements of each module of the device, ensuring a stable output voltage and providing reliable power support for the heating cylinder 3, the single-chip microcomputer module 11, the servo motor module 12, and other components.

[0032] Implementation method 2: Figure 5 As shown, a detection method implemented by the analysis device for gene detection according to any one of the first embodiments comprises the following steps: Step 1: Sample Preparation and Reaction Chamber Loading: First, pretreat the nucleic acid sample to be tested. Mix the nucleic acid template, PCR reaction buffer, primers, dNTPs, DNA polymerase, and other reagents in a sterile environment according to the preset recipe to prepare the PCR reaction system. Use a pipette to transfer a fixed amount of the reaction solution to the microfluidic chip. The PCR reaction chamber is then secured using the reaction chamber turntable of the reaction chamber's synchronous belt rotation module, aligning the reaction chamber's central axis perpendicular to the direction of the synchronous belt's rotation to ensure stability during subsequent heating and detection.

[0033] Step 2: Start the microcontroller module, and the system enters a self-test routine. First, the temperature sensor is calibrated, and sensor errors are corrected using a standard temperature source. Next, the servo motor module is reset to its origin, and the encoder feedback signal determines the initial position of the reaction chamber's synchronous belt rotation module. The user enters the PCR reaction program parameters through the host computer software, including the denaturation temperature, annealing temperature, extension temperature, hold time for each stage, and number of cycles. The user also sets the top fan module's wind speed during the heating phase and the auxiliary wind speed during the cooling phase. Step 3: The MCU module sends a positioning command to the servo motor module. The servo motor drives the synchronous belt via the driving wheel 18, aligning the microfluidic chip reaction pool loaded with PCR reaction solution with the fluorescence detection well. After positioning, the lock seat of the top cover locking module is rotated, driving the lock tongue of the lock assembly to engage with the lock seat on the body. The linkage rod rotates with the lock seat to block the infrared emitting tube. At the same time, the hydraulic rod buffer module is compressed and contracted, ensuring that the elastic pressure cover top cover is tightly fitted to the top surface of the reaction chamber, providing conditions for uniform heating. Step 4: High-Temperature Densification and Hot Air Circulation: The microcontroller module controls the activation of the top cover heating element and fan module. The heating power is dynamically adjusted based on the reaction chamber volume and heating rate requirements. Simultaneously, the top fan module operates at a set speed, driving a convection circulation of air within the reaction chamber. A temperature sensor monitors the reaction solution temperature in real time. Once the temperature reaches the denaturation temperature and stabilizes, it is maintained for 10-30 seconds to allow the double-stranded DNA to completely unwind into single strands. The heating element is deactivated and the fan module is activated. The reaction solution temperature drops to the annealing temperature. The microcontroller module dynamically adjusts the cooling power using a PID algorithm to maintain a stable temperature within ±0.3°C of the target value. This temperature is maintained for 20-60 seconds to allow the designed specific primers to bind to the single-stranded DNA template at the complementary sequence region. The heating element is activated again to raise the reaction solution temperature to the extension temperature. The top fan module speed is adjusted to the midpoint of the heating phase to maintain stable airflow within the reaction chamber. This process is maintained for 30-90 seconds. The single-chip microcomputer module dynamically compensates for the power loss of the heating element through real-time temperature data acquisition, ensuring that the temperature fluctuation during the extension phase is less than ±0.2°C, thereby protecting the activity and synthesis efficiency of the DNA polymerase.

[0034] After the extension phase of each PCR cycle, or at a specific stage according to the settings, the fluorescence detection module is activated. Excitation light from the excitation light source is transmitted through the combined end of the bifurcated optical fiber to the common end, from which it enters the detection area, stimulating the fluorescent probe in the reaction solution to produce fluorescence. The emitted fluorescence signal is collected at the common end and transmitted through the bifurcated optical fiber's splitter end to the fluorescence detector. The detector converts the optical signal into an electrical signal, which is then processed in real time by the microcontroller module after A / D conversion. Step 6. Cyclic reaction and dynamic calibration Repeat steps 3 to 5 until the preset number of cycles is reached. Before each cycle starts, the single-chip microcomputer module automatically calibrates the servo motor position to ensure that the reaction hole position is accurately reset; at the same time, according to the temperature response data of the previous cycle, the heating element power and fan speed are adaptively adjusted to compensate for system errors caused by factors such as reaction chamber loss and ambient temperature changes. When the number of cycles reaches the set value, the system automatically enters the termination program. Result analysis and data output The single-chip microcomputer module processes the fluorescence data of all cycles. The test results can be transmitted to the host computer software through communication interfaces such as USB, RS-485, and WiFi to generate a test report containing Ct value, concentration, amplification curve, and melting curve.

[0035] Step 7: After the device is reset and maintenance tests are complete, the microcontroller detects that the temperature has dropped to a safe range. The top cover locking module is unlocked, and the hydraulic rod buffer module assists in smoothly raising the top cover. The system automatically enters standby mode. Regular maintenance procedures include calibrating the temperature sensor, cleaning the filter of the top fan module, and checking the tension of the timing belt by adjusting the tensioner guide to ensure long-term operational stability and detection accuracy.

[0036] Those skilled in the art will understand that the above are only preferred embodiments of the present invention, and the features described in the various embodiments and technical solutions of the present disclosure can be combined or combined in various ways, even if such combinations or combinations are not explicitly described in the present disclosure. They are not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[0037] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they are aware of the basic inventive concepts. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention. Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, the present invention is intended to include such changes and modifications as fall within the scope of the present invention and its equivalents.

Claims

1. An analysis device based on gene detection, characterized in that: The analysis device comprises a synchronous belt reaction chamber turntable module, a reaction chamber module (6), a servo motor module (12), a top cover module, an external support module, a power supply module (14), a top fan module, a fluorescence detection module, a heating middle cylinder (3), a single chip microcomputer module (11) and a body base; The top fan module comprises a fan bracket (1), a top fan body (2), a spoiler module and a wind speed adjustment module; The top cover module comprises a top cover body (4), a sealing arc strip, a heating element and an elastic pressure cover (5); The computer sends instructions to the single-chip microcomputer module (11), and the single-chip microcomputer module (11) controls the temperature in the reaction chamber module (6). The top fan body (2) delivers regular room temperature air, which is heated to the required hot air by the heating wire in the middle heating cylinder (3) of the reaction chamber module (6). The bottom fan evenly disperses the hot air and evenly heats all the reaction holes. The hot air passes through the reaction holes and is discharged from the hollow position of the elastic pressure cover (5) to the reaction chamber, and is discharged from the exhaust channel between the top cover body (4) and the fuselage. The top cover body (4) is connected to the bottom of the fuselage through a hydraulic rod (15). During the reaction, the servo motor module (12) drives the synchronous belt reaction chamber turntable module. The fluorescence detection module is a four-channel multi-wavelength simultaneous detection optical module. The bottom four-channel multi-wavelength simultaneous detection optical module collects fluorescence data of each reaction hole. The single-chip microcomputer module (11) receives and processes the collected fluorescence data of each reaction hole, and outputs and displays the processed fluorescence data through the host computer.

2. The gene detection-based analysis device according to claim 1, characterized in that: The fan bracket (1) is fixedly mounted above the top cover module, and the wind speed regulating device is connected to the top fan body (2).

3. The analysis device based on gene detection according to claim 1, characterized in that A hydraulic rod buffer module is further provided between the top cover body (4) and the fuselage base, and the hydraulic rod buffer module comprises a hydraulic rod (15), a fixing seat and a connecting module (16); the fixing seats are fixedly mounted on the top cover body (4) and the fuselage, respectively, and the two ends of the hydraulic rod (15) are hinged to the bottom fixing seat and the connecting module (16), respectively.

4. The analysis device based on gene detection according to claim 3, characterized in that A top cover locking module is also provided between the top cover body (4) and the fuselage base, and the top cover locking module includes a locking shaft (22), an infrared radiation sensor (23), a linkage rod (24) and a lock assembly (25); the lock assembly (25) includes a lock tongue and a lock seat; The locking shaft (22) is fixed to the inner side of the top cover body (4) and is connected to the top cover body (4) through a hexagonal nut at the end. One end of the linkage rod (24) is connected to the lock seat, and the other end is inserted into the middle of the infrared radiation sensor (23) to block infrared transmission when the instrument is closed. The lock seat is fixed to the fuselage, and the lock tongue is extended and retracted under the drive of the locking shaft and is engaged with or separated from the lock seat. By operating the servo to rotate the lock seat, the lock seat is driven to engage with the lock tongue, thereby achieving tight locking of the top cover body (4) and the fuselage.

5. The analysis device based on gene detection according to claim 1, characterized in that The synchronous belt reaction chamber turntable module comprises a driving wheel (18), a synchronous transmission belt (19), a rotating table (20), and a tensioning wheel (21). The driving wheel (18) is connected to the output shaft of the servo motor module (12) through a high-precision coupling. The tensioning wheel (21) is installed on the guide rail of the external support module. The engagement between the driving wheel (18) and the synchronous transmission belt (19) drives the synchronous transmission belt (19) to perform a circular motion, and sequentially passes through the detection hole of the fluorescence optical detection module at the bottom of the reaction chamber module (6).

6. The analysis device based on gene detection according to claim 1, characterized in that The servo motor module (12) includes a servo motor, a driver and an encoder; The encoder is installed on the output shaft of the servo motor to detect the rotation position and speed of the servo motor in real time; The signal detected by the encoder is fed back to the driver in the form of pulses, providing the driver with the actual operating status information of the servo motor.

7. The analysis device based on gene detection according to claim 1, characterized in that The sealing arc strip is installed on the edge of the top cover body (4). When the top cover body (4) is matched with the reaction chamber module (6), the elastic pressure cover (5) is used to fill the gap between the top cover body (4) and the reaction chamber module (6) to form a sealing structure. The heating element is realized by a surrounding resistance wire and is evenly laid inside the heating cylinder (3).

8. The gene detection-based analysis device according to claim 1, characterized in that: The external support module comprises a middle support plate (7), a support column (8) and a base plate (10); the base plate (10), the middle support plate (7) and the support column (8) are assembled into a frame structure by bolt connection and corner piece fixation; the support column (8) and the base plate (10) are provided with at least one mounting hole and a guide rail; the mounting hole is used to fix the reaction chamber module (6), the synchronous belt reaction chamber turntable module, the servo motor module (12) and the top cover module; the guide rail is used to provide guidance for the tensioning wheel (21).

9. The gene detection-based analysis device according to claim 1, characterized in that: The power supply module (14) comprises a transformer, a rectifier circuit, a filter circuit and a voltage stabilization circuit, and is used to provide electric energy to the single chip computer module (11), the servo motor module (12) and the heating element.

10. A detection method implemented by the gene detection-based analysis device according to any one of claims 1 to 9, characterized in that: The detection method comprises the following steps: Step 1: First, pre-treat the nucleic acid sample to be tested, use a pipette to transfer a quantitative reaction solution to the microfluidic chip, and fix the PCR reaction chamber through the reaction chamber module (6) and the synchronous belt reaction chamber turntable module so that the central axis of the reaction chamber module (6) is perpendicular to the running direction of the synchronous belt; Step 2: Start the single-chip computer module (11), first calibrate the temperature sensor, and correct the temperature sensor error through the standard temperature source; secondly, reset the servo motor module (12) to its original position, and determine the initial position of the reaction chamber module (6) and the synchronous belt reaction chamber turntable module through the encoder feedback signal; the user inputs the PCR reaction program parameters through the host computer software, including the denaturation temperature, annealing temperature, extension temperature, holding time of each stage and the number of cycles, and at the same time sets the wind speed of the top fan module in the heating stage and the auxiliary wind speed in the cooling stage; Step 3, the single chip computer module sends a positioning instruction to the servo motor module (12), and the servo motor module (12) drives the synchronous belt to operate through the active wheel, and aligns the microfluidic chip reaction pool loaded with PCR reaction solution with the fluorescence detection hole position; after the positioning is completed, the lock seat of the top cover locking module is operated to rotate, and the lock tongue of the lock assembly is driven to engage with the lock seat on the body, and the linkage rod rotates with the lock seat to block the infrared radiating tube, and at the same time, the hydraulic rod buffer module is compressed and contracted to ensure that the elastic pressure cover top cover is tightly fitted with the top surface of the reaction chamber module (6), providing conditions for uniform heating; Step 4, the single chip computer module (11) controls the top cover heating element and the fan module to start, and the heating power is dynamically adjusted according to the volume and heating rate requirements of the reaction chamber module (6). At the same time, the top fan module operates at a set wind speed to drive the air in the reaction chamber module (6) to form a convection circulation; Step 5: Real-time detection of fluorescence signals. After the extension phase of each PCR cycle is completed, the fluorescence detection module is activated. The excitation light emitted by the excitation light source is transmitted to the common end through the combined end of the bifurcated optical fiber. The common end is incident on the detection area, stimulating the fluorescent probe in the reaction solution to produce fluorescence. The emitted fluorescence signal is collected by the same common end and transmitted to the fluorescence detector through the splitting end of the bifurcated optical fiber. The fluorescence detector converts the optical signal into an electrical signal, which is then processed in real time by the single-chip microcomputer module after A / D conversion. Step 6. Repeat steps 3 to 5 until the preset number of cycles is reached. Before each cycle starts, the microcontroller module automatically calibrates the servo motor position to ensure that the reaction well position is accurately reset; at the same time, the heating element power and fan speed are adaptively adjusted according to the temperature response data of the previous cycle; when the number of cycles reaches the set value, the system automatically enters the termination program; the result analysis and data output microcontroller module processes the fluorescence data of all cycles; the test results are transmitted to the host computer software through the USB, RS-485, and WiFi communication interfaces to generate a test report containing the Ct value, concentration, amplification curve, and melting curve.

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