A microfluidic chip nucleic acid constant temperature amplifier for in vitro detection

Abstract

The utility model provides a kind of microfluidic chip nucleic acid thermostatic amplification instrument for in vitro detection.The amplification instrument integrates shell assembly, temperature control component, chip bearing assembly, detection component, control component and user interaction component.The amplification instrument adopts side-illuminating laser detection design, and the chip tray is provided with annular periphery and notch.The laser light source emitter passes through the notch laterally to emit laser light to the detection hole, avoiding bottom pollution and stray light interference, and ensuring independent precision of multi-channel detection.The chip tray is matched with positioning convex groove, gland and stepping motor to realize stable positioning of chip, and the detection error is less than or equal to 2%.The temperature control component realizes accurate temperature control of ±0.5 ℃ by cooperating heating device, temperature control sensor and temperature control mainboard.The equipment integrates sample ID scanning, detection and result viewing one-stop function, and single detection only needs 20 minutes, supports detection of more than 30 kinds of pathogens, has small size and light weight, adapts to on-site rapid detection demand, and greatly improves detection efficiency and accuracy.

Description

Technical Field

[0001] This utility model belongs to the field of nucleic acid detection technology and point-of-care testing (POCT), specifically, it relates to a microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection. Background Technology

[0002] With the widespread application of molecular diagnostic technology, especially the demand for rapid on-site testing, the need for rapid, miniaturized, and high-precision nucleic acid amplification and detection equipment is increasing. Existing molecular diagnostic equipment has the following shortcomings: Firstly, in terms of size, nucleic acid amplification instruments (such as conventional PCR instruments) are relatively large (mostly 400mm×300mm×200mm or larger), occupying significant table space and unsuitable for on-site or point-of-care testing scenarios. Secondly, the temperature control accuracy is insufficient; some small point-of-care testing (POCT) devices use simple heating modules, lacking precise temperature control structures, resulting in amplification temperature fluctuations of ±1℃ or more, affecting detection accuracy. Thirdly, chip positioning is unstable; the microfluidic chip is often placed manually without a precise positioning structure, making it prone to optical detection errors due to chip misalignment. Furthermore, the integration of diagnostic functions is low, requiring additional scanning equipment to input sample information and a separate display screen to view results, making the operation cumbersome and unsuitable for efficient on-site testing.

[0003] Therefore, there is an urgent need for a small nucleic acid amplification device that is compact, has precise temperature control, stable chip positioning, and integrated functions to solve the above problems. Utility Model Content

[0004] This invention provides a microfluidic chip-based nucleic acid isothermal amplification instrument for in vitro detection. The amplification instrument integrates a shell assembly, a temperature control assembly, a chip carrier assembly, a detection assembly, a control assembly, and a user interaction assembly. The amplification instrument employs a side-illuminated laser detection design. The chip tray has an annular periphery and a notch. The laser emitter emits a laser beam laterally through the notch, aiming at the detection aperture, avoiding bottom contamination and stray light interference, ensuring independent and accurate multi-channel detection. The chip tray, combined with positioning grooves, a pressure cap, and a stepper motor, achieves stable chip positioning with a detection error ≤2%. The temperature control assembly, through a heating device, a temperature sensor, and a temperature control motherboard, achieves precise temperature control of ±0.5℃. The device integrates sample ID scanning, detection, and result viewing in a one-stop process, requiring only 20 minutes per test. It supports the detection of over 30 pathogens, is small in size and lightweight, and is suitable for rapid on-site testing needs, significantly improving detection efficiency and accuracy.

[0005] This invention provides a microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection. The amplification instrument includes a chip carrier component and a detection component. The chip carrier component includes a chip tray, and the detection component includes a laser light source emitter. The chip tray is provided with a detection hole and an annular periphery. The annular periphery is provided with a notch, and each detection hole corresponds to a notch. The laser light source emitter passes through the notch and emits a laser beam into the detection hole.

[0006] The chip tested by this device is a disk-shaped multi-channel chip. After the chip is placed on the chip tray, each reaction cavity on the chip is aligned longitudinally with each detection hole on the chip tray. The detection holes are aligned laterally with the notches on the annular periphery of the chip tray. As the chip tray rotates, the notches on the chip tray are aligned sequentially with the laser source emitter. The laser is precisely focused on the center of each reaction cavity of the chip through the notches. At the same time, the annular periphery next to the notches can block the laser from affecting adjacent reaction cavities, ensuring that the detection of each reaction cavity is independent and does not affect each other.

[0007] In existing technologies, the laser emitter is often located below the detection port (reaction chamber) of the chip tray (bottom illumination type). This prevents the integration of other functional components into the chip tray, such as a heating device, which greatly limits the integration capabilities of the amplification instrument. Furthermore, this placement method makes detection susceptible to bottom interference. The laser needs to penetrate the bottom of the chip to enter the detection port, but the bottom of the chip directly contacts the chip tray, making it prone to dust and other contaminants, leading to laser attenuation or refraction deviation. Simultaneously, the contact interface between the chip bottom and the chip tray may generate laser reflection / scattering light. This stray light propagates upwards, superimposing on the fluorescence signal and interfering with detector signal recognition (especially for low-concentration samples, potentially leading to false positives or signal distortion). In addition, vertically oriented laser emitters result in a large space occupation, hindering miniaturization.

[0008] This invention places the laser source emitter on the side of the chip tray (side-illuminated type). On the one hand, the side-illuminated type allows for the integration of heating devices at the bottom of the chip tray, significantly improving the automation of the detection process. On the other hand, the laser in the side-illuminated type is incident horizontally from the side, bypassing the contact interface between the chip and the chip tray, thus avoiding interference from bottom contamination, reflected / scattered light, etc. At the same time, the propagation directions of the laser and the fluorescence signal are at a certain angle, resulting in significantly less stray light interference compared to the bottom-illuminated type, making it particularly suitable for the detection of high-sensitivity, low-concentration nucleic acid samples.

[0009] Furthermore, the chip tray has positioning grooves adapted to the matching microfluidic chip, and the chip tray drives the microfluidic chip to rotate for detection.

[0010] In existing technologies, microfluidic chips are mostly placed manually without a precise positioning structure, making them prone to displacement and causing optical detection errors. The positioning grooves in this device enable rapid and precise chip placement, avoiding horizontal displacement.

[0011] Furthermore, the detection assembly also includes a light source bracket, a photoelectric conversion plate, and a photoelectric conversion plate mounting base.

[0012] The light source bracket finely adjusts the laser emission angle, so that the laser beam from the laser source emitter is precisely focused on the center of each reaction cavity of the chip.

[0013] Furthermore, the chip carrier assembly also includes a pressure cap that covers the chip tray.

[0014] Furthermore, the pressure cap is fitted onto the chip tray via a central thread or snap-fit, which can prevent the chip from shaking or tilting during the testing process.

[0015] Furthermore, the chip carrier assembly also includes a stepper motor; the output shaft of the stepper motor is connected to the bottom of the chip tray and drives the chip tray to rotate.

[0016] The stepper motor drives the chip tray to rotate, so the chips pass through the detection area sequentially for inspection. This device, through the cooperation of the positioning protrusion, the pressure cap, and the stepper motor, ensures precise chip positioning, reducing the detection error to ≤2%, and is easy to operate, eliminating the need for repeated manual chip adjustments and improving detection efficiency.

[0017] Furthermore, the amplification instrument also includes a temperature control component; the temperature control component adopts a PID temperature control system, including a heating device, a temperature control motherboard, and a temperature control sensor; the heating device is installed at the bottom of the chip tray, and the temperature control sensor is located between the heating device and the chip tray; the temperature control motherboard is connected to the heating device.

[0018] Nucleic acid amplification reactions are extremely sensitive to temperature. Excessive temperature fluctuations can lead to low amplification efficiency, poor specificity, and even false positive or false negative results. Placing a temperature sensor between the heating device and the chip tray allows for more direct and rapid capture of tray temperature changes. The temperature control component employs a PID temperature control system, which can adjust the output of the heating device in real time to achieve precise and stable temperature control, with a temperature adjustment accuracy within ±0.5℃.

[0019] Some existing POCT devices use simple heating modules, resulting in poor temperature control accuracy and temperature fluctuations exceeding ±1℃. This invention, through the combined structure of a heating device, a temperature sensor, and a temperature control motherboard, achieves temperature regulation accuracy within ±0.5℃, significantly improving detection accuracy.

[0020] Furthermore, the amplification instrument also includes a housing assembly, which includes an instrument base plate; the heating device includes a heating element and a heating element fixing plate; the light source bracket is fixed to the instrument base plate, and the laser light source emitter is mounted on the top of the light source bracket.

[0021] The outer casing provides protection and mounting support for all internal components, ensuring the overall structural stability of the equipment.

[0022] Furthermore, the stepper motor is mounted below the stepper motor mounting plate, which is mounted on the instrument base plate via a fixing post.

[0023] The stepper motor generates vibrations during operation. If the installation is not secure, these vibrations can cause the chip tray to rotate and shift, affecting chip positioning and detection accuracy. The multi-layered fixing structure of the fixing plate and fixing posts disperses the vibrations generated by the stepper motor during operation, preventing them from being transmitted to the chip tray. Simultaneously, it ensures the stepper motor's position is fixed, guaranteeing the stability of the connection between the output shaft and the chip tray.

[0024] Furthermore, the stepper motor is mounted below the stepper motor mounting plate, which is mounted on the instrument base plate via a fixing post.

[0025] Furthermore, the photoelectric conversion plate is fixed to the photoelectric conversion plate fixing seat, and the photoelectric conversion plate fixing seat is embedded in the heating element fixing plate. Spatially, the photoelectric conversion plate fixing seat is coaxially opposite to the laser source emitter.

[0026] Nucleic acid amplification detection relies on fluorescence signal intensity to determine the amplification status and obtain the detection result. A laser source emitter provides a stable detection light source, and a support bracket ensures its fixed position. A photoelectric conversion board converts the optical signal into an electrical signal that is easy for the control components to process, and a mounting bracket for the photoelectric conversion board ensures its stable position and accurate signal acquisition.

[0027] Due to limited internal space, components need to be rationally arranged to achieve a miniaturized design. Embedding the photoelectric conversion board mounting base into the heating element mounting plate makes full use of space and reduces the volume occupied by components. The laser source emitter is coaxially opposite to the photoelectric conversion board, which ensures that the laser accurately illuminates the chip detection area, and the fluorescence signal can be directly and without deviation transmitted to the photoelectric conversion board, avoiding signal loss or offset.

[0028] Furthermore, the amplification instrument also includes a control component, which includes a scanning component; the housing component also includes an instrument housing; the scanning component includes a scanning gun and a scanning support, the scanning gun is mounted on the scanning support, the scanning support is mounted on the inner side wall of the instrument housing, and the scanning end of the scanning gun faces the outside of the amplification instrument.

[0029] The scanning component can automatically scan the sample ID on the microfluidic chip and transmit it to the control component, enabling rapid entry of sample information.

[0030] In existing technologies, nucleic acid testing equipment has low functional integration and requires additional scanning equipment to input sample information, making the operation process cumbersome. This invention integrates the scanning component inside the instrument, which can directly complete sample ID scanning after the chip is placed, realizing the integration of scanning function and testing equipment. No additional equipment is needed, simplifying the operation process. Sample ID scanning, testing, and result viewing are completed in one stop, reducing the operation process by 50%.

[0031] Furthermore, the amplification instrument also includes a user interaction component, which includes a display screen, an indicator light panel, and a standby button; the control component also includes a main control board and a power module.

[0032] The user interaction component enables device operation (standby button start / stop), status viewing (indicator panel), and detection information display (display screen); the main control board is the core control unit of the device, coordinating the work of various components; the power module provides stable power support for all components of the device.

[0033] Furthermore, the main control board is electrically connected to the temperature control motherboard, the photoelectric conversion board, the user interaction component, the power module, and the scanning component, respectively.

[0034] The main control board's centralized control and data transmission of each component ensures that all components work together to complete the entire automated process from sample entry, temperature control, testing to result display.

[0035] Furthermore, the housing assembly also includes a flip cover structure; the flip cover structure includes a flip cover and a flip cover pivot; the flip cover is rotatable about the flip cover pivot.

[0036] The device requires frequent chip placement and removal, and the flip-top structure facilitates operation. At the same time, it is necessary to maintain a stable internal environment during the testing process to avoid external factors such as dust and light affecting the test results. Closing the flip-top can create a relatively sealed space, ensuring a stable testing environment.

[0037] Flip-top covers need to be opened and closed frequently. The hinge structure can reduce friction between the flip cover and the outer shell, extending its service life. The rotatable design allows users to adjust the flip angle according to their operational needs, improving ease of operation. At the same time, the hinge can limit the opening and closing range of the flip cover, preventing damage to components from excessive opening and closing.

[0038] Furthermore, the amplification instrument is small in size and lightweight.

[0039] In some configurations, the amplification instrument has overall dimensions of 245mm × 200mm × 145mm (L × W × H) and a net weight of 4kg, making it suitable for workbench spaces with limited space and allowing for single-person transport.

[0040] In existing technologies, conventional PCR instruments are large and heavy, making them unsuitable for on-site testing. The device provided by this invention can be miniaturized and adapted to rapid on-site testing scenarios.

[0041] This utility model has the following beneficial effects: 1. Precise chip positioning: The stepper motor drives the chip tray to move, and together with the positioning protrusion and the pressure cover, it avoids chip misalignment and reduces the detection error to ≤2%; 2. Independent multi-channel detection of the chip: During the rotation of the chip tray, the notch of the chip tray is aligned with the laser source emitter in sequence. The laser is precisely focused on the center of each reaction cavity of the chip through the notch. At the same time, the annular outer periphery next to the notch can block the laser from affecting the adjacent reaction cavities, ensuring that the detection of each reaction cavity is independent and does not affect each other. 3. The laser source emitter is located on the side of the chip tray, and a heating device can be integrated at the bottom of the tray, which greatly improves the automation of the detection process and avoids interference from bottom contamination, reflected / scattered light and stray light, making it suitable for the detection of high-sensitivity, low-concentration nucleic acid samples. 4. High temperature control accuracy: Through the combination of heating device + temperature sensor + temperature control motherboard, the temperature control accuracy is ±0.5℃ and the temperature uniformity is ±0.5℃, which meets the requirements of nucleic acid isothermal amplification; 5. High degree of functional integration: It integrates scanning, temperature control, detection, and display functions, requiring no additional equipment. Sample ID scanning, detection, and result viewing are all completed in one stop, simplifying the operation process by 50%. 6. High detection efficiency: Single test time is 20 minutes, supports the detection of more than 30 pathogens, and is suitable for the needs of rapid on-site diagnosis; 7. Compact design: The equipment is small in overall size and lightweight, making it suitable for workbench spaces with limited space and allowing for single-person transport. Attached Figure Description

[0042] Figure 1 This is a front view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 2 This is a top view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 3 This is a left view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 4 This is a right view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 5 This is a rear view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 6This is a bottom view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 7 This is an exploded view of the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 1; Figure 8 This is a schematic diagram of the outer casing assembly in Example 1; Figure 9 This is a schematic diagram of the flip-top structure in Example 1; Figure 10 This is a schematic diagram of the structure of the chip carrier component, temperature control component, and detection component in Example 1; Figure 11 This is an exploded view of the chip carrier component, temperature control component, and detection component in Example 1; Figure 12 This is a schematic diagram of the chip tray structure in Example 1; Figure 13 This is a top view of the chip tray in Example 1; Figure 14 This is a side view of the cap of Example 1; Figure 15 This is a top view of the control component of Embodiment 1; Figure 16 This is a schematic diagram of the control component in Example 1; Figure 17 This is a schematic diagram of the scanning component in Example 1; Figure 18 This is a rear view of the instrument casing in Example 1; Figure 19 This is a test report for detecting human respiratory diseases using the RPD15 chip in an in vitro microfluidic chip nucleic acid isothermal amplification instrument, as described in Example 2. Figure 20 This is the test report of the human HPV detection chip used in the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection in Example 3.

[0043] Figure Descriptions: 1-Outer shell, 2-Instrument base plate, 3-Flip cover structure, 4-Flip cover, 5-Flip cover hinge, 6-Flip cover upper jaw, 7-Flip cover lower jaw, 8-Hinge wire guide sleeve, 9-Base pad, 10-Chip tray, 11-Clip cover, 12-Stepper motor, 13-Groove, 14-Stepper motor mounting plate, 15-Fixing post, 16-Mechanical base plate, 17-Heating element, 18-Heating element mounting plate, 19-Light source bracket, 20-Photoelectric conversion board mounting base, 21-Main control board, 22-Reserved network port, 23-USB port, 24-Power socket, 25-Scanner, 26-Scanner bracket, 27-Display screen, 28-Indicator light panel, 29-Standby button, 30-Display screen bracket, 31-Detection hole, 32-Annular periphery, 33-Notch. Detailed Implementation

[0044] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be noted that the embodiments described below are intended to facilitate the understanding of the present invention and do not constitute any limitation thereof.

[0045] Example 1: A microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection like Figures 1-6 As shown, this embodiment provides a microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection. The amplification instrument includes a shell assembly, a temperature control assembly, a chip carrier assembly, a detection assembly, a control assembly, and a user interaction assembly.

[0046] like Figures 7-9 As shown, the outer casing assembly provides protection and sealing for the internal devices of the amplification instrument, including an instrument housing 1, an instrument base plate 2, and a flip-top structure 3. The flip-top structure 3 consists of a flip cover 4 and a flip-top pivot 5. The flip cover 4 includes an upper flip-top jaw 6 and a lower flip-top jaw 7. The two ends of the flip-top pivot 5 are respectively connected to the upper flip-top jaw 6 and the lower flip-top jaw 7. The flip cover 4 can rotate 0-90° around the flip-top pivot 5. A pivot cable sleeve 8 is fitted on the outside of the flip-top pivot 5 to protect the internal wiring. The four corners of the bottom of the instrument base plate 2 are fixed with base feet 9 to prevent the equipment from sliding.

[0047] like Figure 7 , 10 As shown in Figure 14, the chip carrier assembly enables precise positioning of the microfluidic chip, including a chip tray 10, a pressure cap 11, and a stepper motor 12 (e.g., model 28BYJ-48). The chip tray 10 has a positioning groove 13 (size matching the chip's outer diameter, depth 2-3mm), a detection hole 31, and an annular periphery 32 adapted to the matching microfluidic chip. The annular periphery 32 has a notch 33, with each detection hole 31 corresponding to one notch 33. The groove on the microfluidic chip that adapts to the positioning groove 13 has a notch, which matches a protrusion on the groove 13 (this structure is not labeled in the figure provided in this embodiment), used to align the reaction chamber on the microfluidic chip with the detection hole 31. The pressure cap 11 covers the chip tray 10, and the pressure cap 11 is engaged with the chip tray 10 via a central thread or a snap-fit. In this embodiment, the pressure cap 11 is engaged with the chip tray 10 via a central thread. Figure 14 The output shaft of stepper motor 12 is connected to the bottom of chip tray 10, driving chip tray 10 to rotate, thereby initiating the detection reaction and ensuring precise alignment between the microfluidic chip and the detection component. Stepper motor 12 is mounted below stepper motor mounting plate 14, which is fixed to the mechanism base plate 16 by fixing posts 15 (e.g., 10mm high). The mechanism base plate 16 is fixed to the instrument base plate 2.

[0048] The positioning groove 13 cooperates with the pressure cap 11 for positioning, which can prevent the microfluidic chip from shifting when the stepper motor 12 drives the chip tray 10 to move, reducing the detection error to ≤2%. The stepper motor 12 will generate a certain amount of vibration when running. If the installation is not secure, the vibration will cause the chip tray 10 to rotate and shift, affecting the chip positioning and detection accuracy. Through the multi-layer fixing structure of the stepper motor fixing plate 13 and fixing post 15, the vibration of the stepper motor during operation can be distributed, preventing the vibration from being transmitted to the chip tray, while ensuring that the stepper motor position is fixed and ensuring the stability of the connection between the output shaft and the chip tray.

[0049] The amplification instrument detects a disk-shaped multi-channel chip (in this embodiment, the chip tray 10 has 31 detection holes 31, and the corresponding chip has a maximum of 31 reaction chambers). After the chip is placed on the chip tray 10, each reaction chamber on the chip is aligned longitudinally with each detection hole 31 on the chip tray 10, and the detection hole 31 is aligned laterally with the notch 33 of the annular periphery 32 of the chip tray 10. During the rotation of the chip driven by the chip tray 10, the notch 33 of the chip tray 10 is aligned sequentially with the laser source emitter. The laser is precisely focused on the center of each reaction chamber of the chip through the notch 33. At the same time, the annular periphery 32 next to the notch 33 can block the laser from affecting the adjacent reaction chambers, ensuring that the detection of each reaction chamber is independent and does not affect each other.

[0050] like Figure 7 , 10 As shown in Figure 11, the temperature control component ensures precise and stable amplification temperature, including a heating device, a temperature control mainboard, and a temperature sensor (e.g., NTC type, accuracy 0.1℃). The heating device includes a heating element 17 (e.g., 50mm × 30mm, 10W power) and a heating element fixing plate 18. The heating element 17 is attached to the upper surface of the heating element fixing plate 18; the heating element fixing plate 18 is installed at the lower part of the chip tray 10; the temperature sensor is embedded in the gap between the heating element 17 and the chip tray 10, and in this embodiment, the temperature sensor is embedded in a groove on the upper surface of the heating element 17; the temperature control mainboard is connected to the heating element 17 via a ribbon cable, and achieves temperature regulation accuracy within ±0.5℃ through PID temperature control logic, with a temperature control range of 37℃-60℃. Nucleic acid amplification reactions are extremely sensitive to temperature; excessive temperature fluctuations can lead to low amplification efficiency, poor specificity, and even false positive or false negative results. This device places the temperature sensor between the heating element 17 and the chip tray 10, which can capture the temperature changes of the chip tray 10 more directly and quickly; at the same time, it adopts a PID temperature control system, which can adjust the output of the heating device in real time to achieve precise and stable temperature control.

[0051] like Figure 7 , 10As shown in Figure 11, the detection assembly realizes the emission of laser light source and the acquisition and conversion of fluorescence signals. It includes a light source bracket 19, a laser light source emitter, a photoelectric conversion plate (such as model S11259-01), and a photoelectric conversion plate mounting base 20. The light source bracket 19 is fixed to the instrument base plate 2 by a support column (such as 40mm in height, the support column of the light source bracket 19 is not shown in the figure). The laser light source emitter is installed on the top of the light source bracket 19. By finely adjusting the angle of the light source bracket 19, the laser emitted by the laser light source emitter can pass through the notch 33 and be aligned with the center of the reaction cavity of the chip placed on the detection hole 31. The photoelectric conversion plate is fixed to the photoelectric conversion plate mounting base 20. The photoelectric conversion plate mounting base 20 is embedded in the heating plate mounting plate 18. Spatially, the photoelectric conversion plate mounting base 20 is coaxial with the laser light source emitter and is used to receive fluorescence signals and convert them into electrical signals.

[0052] The amplification instrument places the laser source emitter on the side of the chip tray 10 (side-illuminated). On the one hand, side-illuminated design allows for the integration of heating devices at the bottom of the chip tray 10, significantly improving the automation of the detection process. On the other hand, the side-illuminated laser is incident horizontally from the side, bypassing the contact interface between the chip and the chip tray 10, thus avoiding interference from bottom contamination, reflected / scattered light, etc. At the same time, the propagation directions of the laser and fluorescence signal are at a certain angle, resulting in significantly less stray light interference compared to bottom-illuminated design, making it particularly suitable for the detection of high-sensitivity, low-concentration nucleic acid samples.

[0053] like Figure 7 , 15 As shown in Figure 18, the control component enables the coordinated operation of all components, including a main control board 21, a power module, a scanning component, a reserved network port 22, and a USB port 23. The main control board 21 is fixed to the instrument base plate 2 with screws and is electrically connected to the temperature control motherboard, stepper motor 12, photoelectric conversion board, user interaction component, power module, and scanning component. The power module includes a power socket 24 and a DC power conversion unit. The power socket 24 is located on the back of the instrument housing 1. The power socket 24 receives AC100-240V 50 / 60Hz input, and the conversion unit outputs DC24V 4A to the main control board 21. The scanning component includes a scanner 25 and a scanner holder 26. The scanner 25 is mounted on the scanner holder 26, which is mounted on the inner wall of the instrument housing 1. The scanning end of the scanner 25 faces the outside of the amplification instrument and is used to scan the sample ID and transmit it to the main control board 21. Alternatively, the sample ID can be manually entered without scanning it using the scanner 25. In this embodiment, the scanner bracket 26 is installed on the inner left side of the instrument housing 1, and the reserved network port 22 and USB port 23 are located on the back of the instrument housing 1.

[0054] like Figure 7 , 15As shown in Figure 18, the user interaction components enable operation and result viewing, including a display screen 27, an indicator panel 28, and a standby button 29. The display screen 27 is fixed to the front inclined panel of the instrument housing 1 via a display screen bracket 30, and is used to display detection parameters, progress, and results. In this embodiment, the display screen 27 is a 7-inch 65K color LED display with a resolution of 1024×600. The indicator panel 28 includes three LEDs, fixed to the front of the instrument housing 1, and indicates the device status (standby - green, detection in progress - yellow, abnormal - red) using red / green / yellow lights. The standby button 29 is located on the back of the instrument housing 1 and is electrically connected to the main control board 21 for starting and stopping the device.

[0055] The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection provided in this embodiment has an overall size of 245mm×200mm×145mm (L×W×H) and a net weight of 4kg. It is suitable for limited work surface space and can be carried by a single person.

[0056] The operating procedure for the microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection is as follows: 1. Open the flip cover 4, open the pressure cover 11, place the matching microfluidic chip into the positioning groove 13 of the chip tray 10, and close the pressure cover 11; press the standby button 29, the device is powered on, and the display screen 27 displays the preheating interface (e.g., the target temperature for target nucleic acid amplification is 46.5℃). 2. The temperature control main board controls the heating element 17 to heat up to the target temperature, and the temperature sensor provides real-time temperature feedback to maintain a constant temperature; 3. The barcode scanner 25 scans the sample ID, and the sample ID is transmitted to the main control board 21, or the sample ID can be manually entered on the display screen 27; 4. Start the detection experiment. The main control board 21 controls the stepper motor 12 to drive the chip tray 10 to rotate. At the same time, it can also align the chip detection area with the laser source emitter and photoelectric conversion board to detect the fluorescence intensity. 5. The main control board 21 processes the signal and judges the test result (positive / negative). The display screen 27 displays the result and the test curve. After the test is completed, the indicator light board 28 lights up green, prompting the microfluidic chip to be removed.

[0057] Example 2: Detection of human respiratory diseases using an in vitro microfluidic chip nucleic acid isothermal amplification instrument running an RPD15 chip. 1. Adapting chips and detection targets The microfluidic chip adapted in this embodiment is the RPD15 human respiratory pathogen detection microfluidic chip (invention patent number CN222152080U) independently developed by Suzhou Xianda Gene Technology Co., Ltd. This chip is designed with multi-channel reaction chambers for 15 common respiratory pathogens (including influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, mycoplasma pneumoniae, etc.). Each reaction chamber is preloaded with specific primers, probes and isothermal amplification reagents for the corresponding pathogen. The overall size of the chip is 75mm in diameter and 3mm in thickness. There is a 15mm diameter hole in the center and a groove that can be precisely matched with the positioning protrusion of the chip tray of the amplification instrument.

[0058] 2. Equipment parameter debugging Temperature control parameter settings: Based on the isothermal amplification requirements of the RPD15 chip (the optimal temperature for target nucleic acid amplification is 42℃), the target temperature of the temperature control component is set to 42℃ through the main control board, the temperature control accuracy is maintained within ±0.5℃, and the temperature uniformity is controlled within ±0.5℃ to ensure that the temperature of the 31 reaction chambers is consistent and to avoid amplification efficiency deviations caused by local temperature differences.

[0059] Detection component calibration: The laser source emitter uses a 490nm wavelength FAM channel light source (compatible with the fluorescent markers of the RPD15 chip probes). The laser emission angle is finely adjusted by the light source bracket to ensure that the beam is precisely focused on the center of each reaction chamber of the chip. The signal acquisition threshold of the photoelectric conversion board is set to 500mV to ensure accurate capture of fluorescent signals of low concentrations of pathogens, while filtering background noise and controlling the detection error to ≤2%.

[0060] Stepper motor stroke setting: Since the RPD15 chip uses 31 reaction chambers arranged in a ring, the main control board presets the stepper motor's rotation stroke to "36° per second". The drive chip tray sequentially sends each reaction chamber to the laser detection area to achieve one-by-one scanning of 15 pathogens. The single scan cycle is 10 seconds to ensure that the detection time of each reaction chamber is consistent.

[0061] 3. Actual testing process Sample preparation and loading: Take a nasopharyngeal swab sample from the patient, and after completing the nucleic acid extraction according to the instructions of the sample processing kit for the Suzhou Xianda Gene RPD15 chip, add 660μL of the extraction solution to the chip sample loading well.

[0062] Instrument preparation and preheating: Press the standby button 29 to power on the device. The display screen 27 shows the preheating interface. The temperature control component starts heating and heats up to the target temperature (42℃) within 30 seconds and maintains a constant temperature. The indicator light 28 lights up yellow, indicating that the device has entered the detection preparation state. 4. Equipment Operation Procedures Open the amplification instrument cover 4, place the RPD15 chip loaded with the sample into the chip tray 10, and ensure that the groove of the chip engages with the positioning protrusion 13 in the chip tray 10. Tighten the cover 11 to ensure that the chip is in full contact with the bottom of the chip tray 10.

[0063] Click the "New Detection" button on display screen 27 to start the program. The main control board controls the stepper motor to drive the chip tray to rotate. First, the reagents are mixed at a low speed, and the sample enters the reaction chamber for amplification. After 10 minutes, the rotation speed is increased again, and the reagents enter the detection chamber for fluorescence detection. The laser light source emitter emits a beam, and the photoelectric conversion board synchronously collects the fluorescence signals from each reaction chamber. The signals are converted and transmitted to the main control board for data analysis.

[0064] Results Output and Interpretation: Ten minutes after the detection starts, the main control board 21 completes the fluorescence signal analysis of 15 pathogens and generates a detection report on the display screen 27, including the "positive / negative" result of each pathogen and the corresponding fluorescence intensity curve (the horizontal axis is the detection time and the vertical axis is the fluorescence intensity). If the fluorescence intensity of a pathogen's reaction chamber exceeds the threshold, it is judged as positive, and a red "+" is marked at the corresponding position on the display screen 27. A negative result is marked with a green "-". After the detection is completed, the indicator light panel 28 lights up green, prompting the removal of the chip. The detection data can be displayed on the instrument's display screen 27 or on the connected computer software.

[0065] Test report such as Figure 19 As shown, 5. Application Scenarios and Advantages This embodiment is applicable to on-site testing scenarios such as hospital emergency rooms, fever clinics, and community health service centers. Especially during peak seasons for respiratory diseases (such as the winter and spring flu season), it can complete the entire process of "sample collection-testing-reporting" within 20 minutes. Compared with traditional PCR instruments (testing time of 1-2 hours, requiring a laboratory environment), it significantly shortens the testing cycle, helping to facilitate rapid clinical diagnosis and medication guidance. At the same time, the device is only 245mm×200mm×145mm (L×W×H) in size and weighs 4kg. It can be placed on the examination table in the outpatient clinic without occupying additional laboratory space. It can be moved by a single person, meeting the on-site testing needs of primary healthcare institutions.

[0066] Example 3: Running a human HPV detection chip in a microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection. 1. Adapting chips and detection targets The microfluidic chip adapted in this embodiment is the HPV20 human papillomavirus detection microfluidic chip from Suzhou Xianda Gene Technology Co., Ltd. This chip covers 20 HPV subtypes (including high-risk subtypes 16 and 18 and low-risk subtypes 6 and 11), and adopts a design of "2-3 reaction chambers for important subtypes and 1 reaction chamber for general subtypes". Each reaction chamber is preloaded with subtype-specific primers, TaqMan probes and isothermal amplification buffer. The chip size is 75mm in diameter and 3mm in thickness. The chip is perfectly matched with the positioning groove of the amplification instrument chip tray. The chip has a chip cap that is threaded to the chip tray, pressing the chip tightly onto the chip tray to fix the chip and ensure full contact to improve the efficiency of heating and insulation.

[0067] 2. Equipment parameter debugging Temperature control parameter optimization: The optimal temperature for HPV nucleic acid amplification is 42℃. The target temperature of the temperature control component is set to 42℃ through the main control board. Relying on the heating element, NTC temperature sensor (accuracy 0.1℃) and PID temperature control logic of the temperature control motherboard, a temperature control accuracy of ±0.5℃ is achieved to ensure that the temperature deviation of all reaction chambers does not exceed 0.5℃, avoiding false negative or false positive results caused by temperature fluctuations.

[0068] Precise positioning by stepper motors: The 31 reaction chambers of the HPV20 chip are arranged in a ring (the center of the circle coincides with the center of the chip tray). The main control board presets the rotation angle of the stepper motor to one revolution every 10 seconds, driving the chip tray to rotate at a constant speed, so that each reaction chamber passes through the laser detection area in sequence, and completes the signal acquisition of 20 subtypes in a single detection cycle.

[0069] 3. Actual testing process Sample preparation and loading: Cervical exfoliated cell samples were collected from women and processed using the sample preservation solution for HPV testing provided by Suzhou Xianda Gene. After nucleic acid extraction, 660 μL of the extract was added to the sample loading well of the HPV20 chip. The sample was evenly distributed into the channels corresponding to the 31 reaction chambers through the microchannels inside the chip.

[0070] 4. Equipment Operation Procedures Open the flip cover 4 of the amplification instrument, place the HPV20 chip into the positioning groove 13 of the chip tray 10, and tighten the pressure cap 11 to ensure that the bottom of the chip is in close contact with the heating element (to avoid uneven temperature conduction).

[0071] Press the standby button 29 to power on the device. The temperature control component starts heating up. Once it enters a constant temperature state, the yellow light on the indicator panel 28 will illuminate.

[0072] Click "New Detection", select "HPV20" as the detection item, and enter / scan the information of the sample to be tested. The main control board controls the stepper motor 12 to drive the chip tray 10 to rotate, and the detection program begins.

[0073] Results Output and Interpretation: After 20 minutes of testing, the main control board completes the fluorescence signal analysis of 20 HPV subtypes and generates an "HPV Subtype Detection Report" on the display screen 27. At the same time, a fluorescence intensity curve is generated to help clinicians judge the amplification efficiency. After the test is completed, the green light on the indicator panel 28 will light up. The results can be viewed directly on the instrument display screen 27 or on the computer software, and a report file in your own format can be generated (computer software only). The test data supports long-term storage.

[0074] Test report such as Figure 20 As shown.

[0075] 5. Application Scenarios and Advantages This embodiment is applicable to scenarios such as hospital gynecology departments, maternal and child health care centers, and physical examination centers. Addressing the needs of HPV infection screening, it achieves "rapid results in 20 minutes," significantly improving testing efficiency compared to traditional HPV testing methods (such as real-time fluorescence PCR, which takes over 1.5 hours). Simultaneously, the device integrates scanning, temperature control, detection, and result display functions, eliminating the need for additional scanning instruments or computers; a single person can operate it, lowering the operational threshold for primary healthcare institutions. Furthermore, the compatibility between Suzhou Xianda Gene's HPV20 chip and this amplification instrument has been rigorously verified, achieving a detection accuracy rate of over 98%, meeting the clinical demand for precise HPV genotyping detection and aiding in early cervical cancer screening and intervention.

[0076] While the present invention has been disclosed above, it is not limited thereto. Any person skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention; therefore, the scope of protection of the present invention should be determined by the scope defined in the claims.

Claims

1. A microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection, characterized in that, The amplification instrument includes a chip carrier component and a detection component; the chip carrier component includes a chip tray, and the detection component includes a laser source emitter; the chip tray is provided with a detection hole and an annular periphery, and the annular periphery is provided with a notch, with each detection hole corresponding to a notch; the laser source emitter passes through the notch and aims at the detection hole to emit a laser.

2. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 1, characterized in that, The chip tray has positioning grooves that are compatible with the matching microfluidic chip. The chip tray drives the microfluidic chip to rotate for detection.

3. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 2, characterized in that, The chip carrier assembly also includes a pressure cap that covers the chip tray.

4. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 3, characterized in that, The amplification instrument also includes a temperature control component; the temperature control component adopts a PID temperature control system, including a heating device, a temperature control motherboard and a temperature control sensor; the heating device is installed at the bottom of the chip tray, and the temperature control sensor is located between the heating device and the chip tray; the temperature control motherboard is connected to the heating device.

5. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 4, characterized in that, The amplification instrument also includes a housing assembly, which includes an instrument base plate; the heating device includes a heating element and a heating element fixing plate; the light source bracket is fixed to the instrument base plate, and the laser light source emitter is mounted on the top of the light source bracket.

6. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 5, characterized in that, The photoelectric conversion plate is fixed to the photoelectric conversion plate fixing seat, which is embedded in the heating element fixing plate. Spatially, the photoelectric conversion plate fixing seat is coaxial with the laser source emitter.

7. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 6, characterized in that, The amplification instrument also includes a control component, which includes a scanning component; the housing component also includes an instrument housing; the scanning component includes a scanning gun and a scanning support, the scanning gun is mounted on the scanning support, the scanning support is mounted on the inner side wall of the instrument housing, and the scanning end of the scanning gun faces the outside of the amplification instrument.

8. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 7, characterized in that, The amplification instrument also includes a user interaction component, which includes a display screen, an indicator light panel, and a standby button; the control component also includes a main control board and a power module.

9. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 8, characterized in that, The main control board is electrically connected to the temperature control motherboard, photoelectric conversion board, user interaction component, power module and scanning component respectively.

10. The microfluidic chip nucleic acid isothermal amplification instrument for in vitro detection as described in claim 9, characterized in that, The outer casing assembly also includes a flip cover structure; the flip cover structure includes a flip cover and a flip cover pivot; the flip cover is rotatable about the flip cover pivot.

Images