Handheld variable-temperature high-flux nucleic acid amplification detector and application thereof

Abstract

The invention relates to a handheld variable-temperature high-flux nucleic acid amplification detector and application thereof. The detection module of the handheld variable-temperature high-flux nucleic acid amplification detector comprises a light path module structure, the light path module structure is detachably installed on a light path support through an exciting light assembly and an emitting light assembly, the light path support is a polygon frustum and comprises a top face, a bottom face and side faces, the top face and the bottom face are parallel to a chip, and the side faces are parallel to the chip. More than one side face is arranged to be an inclined face, one exciting light assembly is arranged in the center of each inclined face, and the inclination of the inclined faces and the installation angles of the exciting light assemblies are matched to ensure that the angle of light of each exciting light assembly entering the chip ranges from 30 degrees to 41 degrees. The multiple modules are independent and do not influence one another, detection can be carried out at any time, waiting is not needed, the system is very suitable for being used for pathogen site instant detection, the multiple modules are controlled through one mobile phone Bluetooth, operation is easy, and result analysis can be obtained within a short time.

Description

Technical Field

[0001] The present invention relates to the field of biology and medical detection technology, and in particular to a handheld variable temperature high-throughput nucleic acid amplification detector and applications thereof. Background Art

[0002] Real-time fluorescence-based in vitro nucleic acid amplification techniques are divided into temperature-dependent polymerase chain reaction (PCR) and isothermal nucleic acid amplification. These techniques utilize in vitro provision of the necessary raw materials for specific nucleic acid amplification, such as primers, probes / dyes, enzymes, and dNTPs, while maintaining a suitable temperature. Amplification results are then analyzed using fluorescence signal detection. Traditional PCR testing typically requires samples to be taken back to the laboratory for processing and analysis, which prolongs the time it takes to obtain results. Existing equipment is mostly desktop laboratory-based nucleic acid amplification analyzers, utilizing embedded systems or computer control. This makes them difficult to implement for on-site testing and unsuitable for on-demand testing. Handheld variable-temperature PCR detectors, on the other hand, enable immediate PCR reactions and result analysis in the field or clinical setting, enabling immediate testing and diagnosis. These devices have broad application prospects in clinical diagnosis, food safety monitoring, environmental monitoring, drug development, and other fields, meeting the needs of diverse PCR testing applications. They typically offer rapid detection speeds, completing PCR reactions and result analysis within tens of minutes. Therefore, there is a need to develop more practical handheld variable-temperature PCR detectors to meet market demand.

[0003] Currently, most nucleic acid amplification detection devices use real-time fluorescence detection. Chinese Patent 202311391103.0 discloses a handheld nucleic acid detection device, and Chinese Patent 202321540008.8 discloses a portable real-time fluorescence isothermal nucleic acid detection system. These are all based on traditional PCR to detect nucleic acid reagent tubes. The shortcomings of existing handheld PCR detectors are: (1) they are not small enough; (2) the throughput is not high enough, and they cannot be tested on demand; (3) they are not flexible and scalable, and the number of detection modules cannot be increased or decreased according to demand. With the development of microfluidic chip detection technology, if a handheld PCR detector using a microfluidic chip as a carrier is developed, the problems of the existing technology can be solved. Summary of the Invention

[0004] In order to solve the above problems, the present invention proposes a handheld variable temperature high-throughput nucleic acid amplification detector and its application.

[0005] The present invention discloses a handheld variable temperature high-throughput nucleic acid amplification detector, which consists of a power supply module, multiple detection modules and a control module. The detection module includes an optical path module structure, a heating module structure, a chip clamping module structure and a rapid heat dissipation structure. A chip is arranged on one side of the optical path module structure, and the chip is pressed against the heating module structure by the chip clamping module structure. The rapid heat dissipation structure is arranged on the other side of the heating module structure; the optical path module structure is detachably mounted on an optical path bracket by an excitation light component and an emission light component, and the optical path bracket is a multi-faceted pyramid, including a top surface, a bottom surface and side surfaces. The top surface and the bottom surface are parallel to the chip, and more than one side surface is set as an inclined surface. An excitation light component is set in the center of each inclined surface, and the inclination of the inclined surface and the installation angle of the excitation light component are matched to ensure that the angle of light from each excitation light component entering the chip is 30-41 degrees. ; The excitation light component is composed of an excitation LED lamp and an excitation light filter, and the excitation light filter is detachably mounted on the excitation light filter positioning groove set on the side of the optical path bracket, and the excitation LED lamp is detachably mounted on the excitation light filter positioning structure, and the excitation light filter positioning structure is detachably mounted on the excitation light filter positioning groove, and the bottom edge of the excitation light filter positioning structure presses the periphery of the excitation light filter; the emission light component is composed of a photodetector and an emission light filter, and the emission light filter is detachably mounted on the emission light filter positioning groove set on the top surface of the optical path bracket, and the photodetector is detachably mounted on the emission light filter positioning structure, and the emission light filter positioning structure is detachably mounted on the emission light filter positioning groove, and the bottom edge of the emission light filter positioning structure presses the periphery of the emission light filter.

[0006] Furthermore, the angle at which the light of each excitation light component enters the chip is 41 degrees.

[0007] Furthermore, protrusions are provided on both sides of the excitation light filter positioning structure, LED lamp positioning grooves are provided on both sides of the excitation light filter positioning groove, and the protrusions are snap-fitted into the LED lamp positioning grooves.

[0008] Furthermore, an LED positioning screw hole is provided on the inclined surface of the optical path bracket at a position corresponding to the excitation LED lamp chassis, and the excitation LED lamp chassis is pressed against the excitation light filter positioning structure by screw locking, and the excitation light filter is further pressed against the excitation light filter positioning groove.

[0009] Furthermore, the heating module structure includes a silicon-based heating plate, a digital temperature sensor, and a heater circuit substrate. The silicon-based heating plate is welded to the heater circuit substrate through a welding port, and the digital temperature sensor is fixed on a limit slot 1 of the heater circuit substrate. The digital temperature sensor is partially fitted with the silicon-based heating plate.

[0010] Furthermore, a second limiting slot is provided on the heater circuit substrate.

[0011] Furthermore, the chip clamping module structure includes a circular spring, a pressing device, and a fixing part. The circular spring is connected to the pressing device through a connecting component. The pressing device is arranged on the fixing part. The pressing device is used to push the circular spring to press and clamp the chip.

[0012] Furthermore, a limit block is provided on the bottom surface of the fixing member, and the limit block is used to prevent the pressing device from pushing the circular spring too far and causing damage to the chip.

[0013] Furthermore, the rapid heat dissipation structure is a heat dissipation fan.

[0014] Furthermore, the power supply module is provided with a plurality of independent power supply ports.

[0015] The handheld variable temperature high-throughput nucleic acid amplification detector provided by the present invention can be used on a mobile phone. The control module is an application software installed on the mobile phone, which is connected to the detection module via Bluetooth. It can control each detection module individually and add new detection modules as needed.

[0016] The beneficial effects of the present invention are as follows:

[0017] This invention proposes a scalable, modular, handheld, variable-temperature, high-throughput PCR instrument capable of simultaneously testing 1-N samples. Multiple modules are independent and mutually exclusive, enabling on-demand testing without waiting, making it ideal for on-site pathogen detection. Multiple modules are controlled via a single mobile phone Bluetooth connection, making operation simple and enabling rapid analysis of results.

[0018] 2. The handheld variable temperature high-throughput nucleic acid amplification detector (handheld variable temperature high-throughput PCR detector) provided by the present invention adopts a new type of optical path module structure, by detachably mounting the excitation light component and the emission light component on the optical path bracket, the optical path bracket is a polygonal pyramid, including a top surface, a bottom surface and a side surface, the top surface and the bottom surface are parallel to the chip, the side surface is an inclined surface, and an excitation light component is arranged in the center of each side surface. The inclination of the side surface and the installation angle of the excitation light component ensure that the angle of each excitation light component entering the chip is 30-41 degrees, preferably 41 degrees. According to the technical solution of the present invention, the volume of the instrument can be reduced while ensuring the quality of real-time fluorescence detection, making the structure of the handheld variable temperature high-throughput PCR detector more compact. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 , schematic diagram of the structure of a handheld variable temperature high-throughput PCR detector;

[0020] Figure 2 , schematic diagram of the optical path module structure;

[0021] Figure 3 、 4 A schematic cross-sectional view of the optical path module structure with an inclined plane;

[0022] Figure 4 , schematic diagram of the heating module structure;

[0023] Figure 5 , schematic diagram of the assembled heating module structure;

[0024] Figure 6 , schematic diagram of the assembled heating module structure and the rapid heat dissipation structure;

[0025] Figure 7 , schematic diagram of the chip clamping module structure;

[0026] Figure 8 , schematic diagram of the power supply module.

[0027] Explanation of the accompanying drawings: 1. Optical path module structure; 2. Heating module structure; 3. Chip clamping module structure; 4. Rapid heat dissipation structure; 5. Excitation LED lamp; 6. Excitation light filter; 7. Excitation light filter positioning structure; 8. Photodetector; 9. Emission light filter positioning structure; 10. Emission light filter; 11. Emission light filter positioning groove; 12. LED positioning screw hole; 13. LED lamp positioning groove; 14. Chip; 15. Excitation light filter positioning groove; 16. Silicon-based heating plate; 17. Digital temperature sensor; 18. Heater circuit substrate; 19. Welding port; 20. Limiting slot 2; 21. Limiting slot 1; 22. Circular spring; 23. Limiting block; 24. Pressing device; 25. Fixing part; 30. Optical path bracket. DETAILED DESCRIPTION

[0028] The present invention will be further described below with reference to the examples.

[0029] Example 1: A handheld variable temperature high-throughput PCR detector:

[0030] like Figure 1 As shown, a handheld variable temperature high-throughput PCR detector consists of a power supply module, multiple detection modules and a control module. The detection module includes an optical path module structure 1, a heating module structure 2, a chip clamping module structure 3, and a rapid heat dissipation structure 4. A chip 14 is set on one side of the optical path module structure 1, and the chip clamping module structure 3 presses the chip 14 against the heating module structure 2. The rapid heat dissipation structure 4 is set on the other side of the heating module structure 2;

[0031] like Figure 2-3 As shown, the optical path module structure 1 is detachably mounted on an optical path bracket 30 by an excitation light component and an emission light component. The optical path bracket 30 is a polygonal pyramid, including a top surface, a bottom surface and a side surface. The top surface and the bottom surface are parallel to the chip 14. More than one side surface is set as a slope, and an excitation light component is set in the center of each slope. The inclination of the slope and the installation angle of the excitation light component ensure that the angle of each excitation light component entering the chip 14 is 41 degrees; the number of slopes can be increased as needed, and the number of excitation light components can also be increased as needed.

[0032] Specifically, such as Figure 2 As shown, the optical path bracket 30 of this embodiment is a quadrangular pyramid containing three inclined surfaces, and an excitation light component is arranged at the center of each inclined surface. The inclination of the inclined surface and the installation angle of the excitation light component ensure that the angle of each excitation light component entering the chip 14 is 41 degrees.

[0033] Figure 3The light path bracket 30 is shown as a quadrangular pyramid, which contains a cross-sectional structural diagram of four inclined planes on the upper, lower, left and right sides. It can be seen that in the light path bracket 30, the inclination of the inclined plane and the installation angle of the excitation light component can be coordinated to ensure that the angle of light from each excitation light component entering the chip 14 is 41 degrees.

[0034] The excitation light component is composed of an excitation LED lamp 5 and an excitation light filter 6, and the excitation light filter 6 is detachably mounted on the excitation light filter positioning groove 15 set on the side of the optical path bracket, and the excitation LED lamp 5 is detachably mounted on the excitation light filter positioning structure 7, and the excitation light filter positioning structure 7 is detachably mounted on the excitation light filter positioning groove 15, and the bottom edge of the excitation light filter positioning structure 7 presses the periphery of the excitation light filter 6; the emission light component is composed of a photodetector 8 and an emission light filter 10, and the emission light filter 10 is detachably mounted on the emission light filter positioning groove 11 set on the top surface of the optical path bracket, and the photodetector 8 is detachably mounted on the emission light filter positioning structure 9, and the emission light filter positioning structure 9 is detachably mounted on the emission light filter positioning groove 11, and the bottom edge of the emission light filter positioning structure 9 presses the periphery of the emission light filter 10.

[0035] Bumps are provided on both sides of the excitation light filter positioning structure 7 , and LED lamp positioning grooves 13 are provided on both sides of the excitation light filter positioning groove 15 . The bumps are snap-fitted into the LED lamp positioning grooves 13 .

[0036] An LED positioning screw hole 12 is set on the inclined surface of the optical path bracket at a position corresponding to the chassis of the excitation LED lamp 5. By tightening the screws, the chassis of the excitation LED lamp 5 is pressed against the excitation light filter positioning structure 7, and the excitation light filter 6 is further pressed against the excitation light filter positioning groove 15.

[0037] The excitation LED lamp 5 has many advantages as a light source, such as high brightness, low energy consumption and long life. The excitation LED lamp 5 is fixed to the LED lamp positioning structure 7 by screwing through the LED positioning screw hole 12, and the lamp is fixed by directly pressing on the excitation light filter 6 to prevent it from falling off during transportation. Light is irradiated onto the chip 14 through the excitation light filter 6. The excitation light filter 6 can selectively filter out light of a specific wavelength, thereby achieving excitation of the sample. By properly selecting the wavelength and characteristics of the filter, it can be ensured that only light that matches the required fluorescence characteristics is transmitted to the sample. Once the sample is excited, the fluorescent substance therein will emit a fluorescent signal. These fluorescent signals need to pass through the filter to ensure that only the fluorescent signal from the sample is transmitted to the next detector. The function of the filter is to remove non-fluorescent signals, such as excitation light or ambient light, thereby improving the signal-to-noise ratio of the detector and enhancing the detectability of the signal. Ultimately, the fluorescence signal passes through emission filter 10 and then reaches photodetector 8, a device that converts light into electrical signals. This device measures and records the fluorescence intensity in the sample. These electrical signals can be amplified and processed by appropriate circuitry to provide information about the concentration and characteristics of the fluorescent substance in the sample.

[0038] During the entire process, the excitation LED lamp 5, the excitation light filter 6, the emission light filter 10, and the photodetector 8 and other components work closely together to form a high-efficiency fluorescence detection system.

[0039] Each excitation LED 5 has a trapezoidal excitation filter positioning slot 15 for placing the excitation filter 6, which is pressed tightly into the excitation filter positioning slot 15 by the excitation filter positioning structure 7. The interior of the slot is a stepped structure. This stepped design can help firmly fix the optical element and ensure its correct position in the light path, providing stable support to prevent the filter from falling off during device vibration and causing detection failure. It also ensures accurate alignment between optical elements and reduces light offset and scattering.

[0040] There is also an emission light filter positioning slot 11 in front of the photodetector 8. This slot also serves to securely hold the optical element and ensure its correct position in the optical path. This slot provides stable support and prevents the filter from falling off during device vibration, leading to detection failure. It also ensures accurate alignment between optical elements, reduces light offset and scattering, and ensures that the detected fluorescence can be received by the photodetector 8. Alternatively, the optical detector can be integrated with the filter, with the filter embedded in the front end of the detector. In this case, the emission light filter positioning slot 11 is not required.

[0041] The optical module structure 1 of the present invention can utilize various filter types in conjunction with light sources and light detectors. This includes single-bandpass filters, multiple light source and light detector combinations, and the ability to filter the light sources through multi-bandpass filters. Multi-bandpass filters can also be used in front of the light detectors to selectively transmit light signals in specific wavelength bands. Furthermore, single-bandpass lenses can be used in front of area or linear array CCDs or CMOS arrays to transmit different colors of light at different locations, achieving spatial resolution of different fluorescence signals. The photodetector 8 utilizes a silicon photodiode to detect light intensity.

[0042] The present invention can be used in conjunction with a lower computer with a certain storage function. The collected real-time fluorescence intensity values ​​can be stored in its own memory and transmitted to the mobile phone after the reaction is completed. In this way, even if the Bluetooth connection between the device and the mobile phone is interrupted during the detection process, the detection can still proceed normally without affecting the reading of the results. The results are analyzed on the mobile phone, and the user can see the positive or negative test results.

[0043] like Figure 4-5 As shown, the heating module structure 2 includes a silicon-based heating plate 16, a digital temperature sensor 17, and a heater circuit substrate 18. The silicon-based heating plate 16 is welded to the heater circuit substrate 18 through a welding port 19. The digital temperature sensor 17 is fixed on a limiting slot 1 21 of the heater circuit substrate 18. The digital temperature sensor 17 is partially fitted with the silicon-based heating plate 16. A limiting slot 20 is provided on the heater circuit substrate 18.

[0044] The silicon-based heating plate 16 can generate heat after being powered on. By tightly fitting the digital temperature sensor 17 on the silicon-based heating plate 16, the current temperature can be accurately obtained to ensure precise control. The heater circuit substrate 18 is fixed in the instrument through the limit slot 20 to prevent loosening during transportation.

[0045] The temperature of the present invention can be set to any temperature between 37-95 degrees Celsius and is compatible with a variety of detection reagents. In addition to the silicon-based heating plate, it can also be an electric heating film or use other heating parts such as Peltier to achieve heating.

[0046] Specifically, the heating module structure 2 uses a silicon-based heating plate 16, and a digital temperature sensor 17 is provided on the silicon-based heating plate. The resistance value of the sensor changes with temperature. The channel control module reads the resistance value and converts it into temperature for real-time temperature detection, thereby achieving accurate temperature control. Compared with traditional heating methods, the present invention uses a heating plate in close contact with the reaction chip to greatly improve the heating rate and shorten the PCR detection time.

[0047] like Figure 7As shown, the chip clamping module structure 3 includes a circular spring 22, a pressing device 24, and a fixing part 25. The circular spring 22 is connected to the pressing device 24 through a connecting component. The pressing device 24 is arranged on the fixing part 25. The pressing device 24 is used to push the circular spring 22 to press and clamp the chip 14.

[0048] A limiting block 23 is provided on the bottom surface of the fixing member 25 , and the limiting block 23 is used to prevent the pressing device 24 from pushing the circular spring 22 too far and causing damage to the chip.

[0049] The circular spring 22 has a certain elasticity and can return to its original shape when compressed or stretched by external force. When installing the chip, it can be clamped in the middle of the target position and leave a circular space to expose the detection area. The locking function of the circular spring 22 is unlocked by the pressing device 24. After pressing, the circular spring 22 can be pushed forward to the position of the limit block 23. The circular spring 22 can contact the chip 14 and continuously provide pressure to press the chip onto the heating plate. After releasing the pressing device 24, the circular spring 22 leaves the chip 14 and slides back to its original position, and the chip 14 can be removed.

[0050] like Figure 6 As shown, the rapid heat dissipation structure 4 is a heat dissipation fan. The present invention sets a heat dissipation fan behind the silicon-based heating plate to quickly cool the heating plate to achieve PCR detection. Compared with similar products, the cooling rate is greatly improved, shortening the PCR detection time.

[0051] like Figure 8 As shown, the power supply module is provided with multiple independent power supply ports.

[0052] This embodiment uses a four-in-one module for power supply: a power adapter with a 220V AC input and an 18V DC output is used. Two buck-type DC-to-DC circuits (Buck circuits) convert the 220V input voltage into four 18V DC channels to power four modules. Each DC-to-DC circuit, comprised of the selected components, can operate at a peak current of up to 5A. Depending on the design of the power supply circuit and the choice of power adapter, up to 20 modules can be used.

[0053] The handheld variable-temperature, high-throughput nucleic acid amplification detector provided in this embodiment comprises a control module, which is a mobile phone application software that connects to the detection modules via Bluetooth. This control module can independently control each detection module and add new detection modules as needed. The PCR detection program can be input through the mobile phone, and all detection modules can be controlled through the mobile phone control app. The mobile phone control app transmits data to each detection module via Bluetooth communication, including the temperature and time for detection, fluorescence detection settings, and fluorescence intensity data transmitted from the device to the mobile phone. The mobile phone app analyzes the test results and displays them to the user.

[0054] When using:

[0055] (1) Turn on the handheld variable temperature high-throughput nucleic acid amplification detector, log in to the APP on your mobile phone, connect the detection device via Bluetooth (pay attention to select the number of the detection module to be used for connection, and do not connect the wrong module), scan the QR code on the test kit, identify the test item, and transmit the detection temperature, time and fluorescence acquisition parameters to the device;

[0056] (2) Click the "Start Detection" button on your phone;

[0057] (3) Collect samples such as throat swabs, nasopharyngeal swabs, and nasal swabs and add them to the extraction-free sample preservation solution;

[0058] (4) Add the sample preservation solution into the reagent tube;

[0059] (5) Shake the reagent tube and pipette the reagent into the detection chip, insert the chip into the device, and close the device cover;

[0060] (6) Click on the mobile phone to start the test. The test starts and the device counts according to the time recognized by the QR code. At the same time, the light source is turned on at a fixed time according to the acquisition parameters recognized by the QR code. At the same time, the photoelectric detector collects the corresponding fluorescence, obtains the fluorescence intensity and records it;

[0061] (7) After the reaction is completed, log in to the mobile phone app again and connect the device via Bluetooth. The test results will be transmitted from the device to the mobile phone via Bluetooth, and the mobile phone will analyze and display the test results (negative / positive);

[0062] (8) After clicking Preheat in step (2), or after clicking Start Detection in step (6), you can return to the Bluetooth connection interface, select another detection module to connect, and start a new detection. This will not affect the detection that has already been carried out.

[0063] How different modules run independently:

[0064] The mobile phone APP can only connect to one module at a time, but the module does not need to maintain Bluetooth connection when running a program. Therefore, you can first send instructions to the detection module through Bluetooth connection, then disconnect and let the detection module run on its own. After the running is completed, reconnect through the Bluetooth module to obtain the results. The specific usage process is as follows:

[0065] (1) Open the mobile phone APP, and multiple detection modules that have been connected to the power supply will be displayed. Select the module number to be run and connect Bluetooth;

[0066] (2) Scan the QR code containing the detection temperature requirements and light signal acquisition requirements, light intensity, exposure time, etc., and transfer the detection program to the memory of the detection module;

[0067] (3) Prepare the reagents and use a pipette to draw up the reagents, load them onto the detection chip, and then place them into the detection module to start the detection process;

[0068] (4) Switch to another detection module, connect Bluetooth, and restart the settings of (2) and (3) to detect other samples;

[0069] (5) After the module's detection indicator lights up, connect the module via Bluetooth on the mobile app to obtain the detection results.

[0070] During the testing process, you can switch Bluetooth connections to different testing modules to obtain the current status of each testing module without affecting its operation or other testing modules. This allows samples to be tested as soon as they arrive, without having to wait for the previous round of testing to end.

[0071] Compared with the existing miniaturized PCR detector using a multi-luminescence system, the present invention has an average fluorescence signal intensity of 2000 AU and an average signal-to-noise ratio of 10:1, while the average fluorescence signal intensity of the existing miniaturized PCR detector is 1900 AU and the average signal-to-noise ratio is 8:1; the present invention has a heating rate of about 16°C / s and a cooling rate of about 3°C / s, while the existing miniaturized PCR detector has a heating rate of about 2°C / s and a cooling rate of about 0.5°C / s; the total PCR detection time of the present invention is 30 minutes, while the total PCR detection time of the existing miniaturized PCR detector is 60 minutes; the total volume of the present invention is 50 cm 3 , while the total volume of existing miniaturized PCR detectors is 150cm 3 It can be seen that the rapid temperature ramp system of the present invention is significantly superior to the existing technology in terms of temperature ramp rate, greatly shortening the PCR detection time. At the same time, while maintaining good fluorescence signal intensity and signal-to-noise ratio, it significantly reduces the volume of the equipment. Compared with existing miniaturized PCR detectors, the volume of the handheld variable temperature high-throughput nucleic acid amplification detector of the present invention has been reduced by 67%.

[0072] Example 2: Verification of the influence of the incident angle of the excitation light source of the handheld variable temperature high-throughput nucleic acid amplification detector:

[0073] To verify the optimal angle of light source in the handheld variable temperature high-throughput nucleic acid amplification detector of the present invention, 3D-printed optical module structures with incident angles of 20, 30, and 41 degrees were used to excite and collect fluorescence from (1) a chip filled with FAM fluorescein; (2) PCR detection and amplification were performed using optical module structures at different angles after loading with COVID-19 standard reagents to check the fluorescence increment and amplification status. The same miniaturized high-throughput nucleic acid amplification detector motherboard power supply and the same specification excitation LED lamp were used in the experiment to ensure consistency of external parameters.

[0074] Equipment: Miniaturized high-throughput nucleic acid amplification detector (ID: 02), 3-color microfluidic reagent chip (P6, P7 concentration), 3 sets of optical path structures with different incident angles.

[0075] The experimental results are shown in Table 1:

[0076] Table 1 Test results of incident angle of excitation light source

[0077] Angle of incidence P6 FAM Incremental P7 FAM Increment P6 CT value P7 CT value 20 degrees 93 48 11.48 13.49 30 degrees 116 116 10.38 12.36 41 degrees 116 118 9.82 11.50

[0078] From the results of the above comparative experiments and Table 1, it can be seen that under the same external conditions, good fluorescence increment and amplification can be obtained at an incident angle of 30-41 degrees for the excitation LED light source, especially at 41 degrees, the fluorescence increment is higher. The higher the fluorescence increment, the better the excitation efficiency, and the CT value after amplification is closer to the front, indicating that the detection sensitivity is higher. Therefore, 41 degrees is the optimal angle. The reason for not using an angle greater than 41 degrees here is that when it exceeds 41 degrees, a larger optical path module structure is required to adapt, which will make the device larger and does not meet the research and development requirements of the present invention.

[0079] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0080] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A handheld variable temperature high-throughput nucleic acid amplification detector, consisting of a power supply module, multiple detection modules and a control module, characterized in that: The detection module comprises an optical path module structure (1), a heating module structure (2), a chip clamping module structure (3), and a rapid heat dissipation structure (4); a chip (14) is arranged on one side of the optical path module structure (1), and the chip clamping module structure (3) presses the chip (14) against the heating module structure (2); the rapid heat dissipation structure (4) is arranged on the other side of the heating module structure (2); the optical path module structure (1) is detachably mounted on an optical path bracket (30) by an excitation light component and an emission light component; the optical path bracket is a multi-prism body, comprising a top surface, a bottom surface and side surfaces; the top surface and the bottom surface are parallel to the chip (14); more than one side surface is arranged as an inclined surface; an excitation light component is arranged at the center of each inclined surface; the inclination of the inclined surface and the installation angle of the excitation light component are matched to ensure that the angle at which the light of each excitation light component enters the chip (14) is 30-41 degrees; the excitation light component is composed of an excitation LED lamp (5) and an excitation light filter (6); The excitation light filter (6) is detachably mounted on an excitation light filter positioning groove (15) provided on the side of the optical path bracket, the excitation LED lamp (5) is detachably mounted on an excitation light filter positioning structure (7), the excitation light filter positioning structure (7) is detachably mounted on the excitation light filter positioning groove (15), and the bottom edge of the excitation light filter positioning structure (7) presses the periphery of the excitation light filter (6); the emission light component is composed of a photodetector (8 ) and an emission light filter (10), the emission light filter (10) being detachably mounted on an emission light filter positioning groove (11) arranged on the top surface of the optical path bracket, the photodetector (8) being detachably mounted on an emission light filter positioning structure (9), the emission light filter positioning structure (9) being detachably mounted on the emission light filter positioning groove (11), and the bottom edge of the emission light filter positioning structure (9) pressing the periphery of the emission light filter (10).

2. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The angle at which the light of each excitation light component enters the chip (14) is 41 degrees.

3. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The excitation light filter positioning structure (7) is provided with protrusions on both sides, the excitation light filter positioning groove (15) is provided with LED light positioning grooves (13) on both sides, and the protrusions are snap-fittedly installed in the LED light positioning grooves (13).

4. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: An LED positioning screw hole (12) is provided on the inclined surface of the optical path bracket at a position corresponding to the chassis of the excitation LED lamp (5), and the chassis of the excitation LED lamp (5) is pressed against the excitation light filter positioning structure (7) by means of screw locking, and the excitation light filter (6) is further pressed against the excitation light filter positioning groove (15).

5. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The heating module structure (2) comprises a silicon-based heating plate (16), a digital temperature sensor (17), and a heater circuit substrate (18); the silicon-based heating plate (16) is welded to the heater circuit substrate (18) via a welding port (19); the digital temperature sensor (17) is fixed to a first limiting card slot (21) of the heater circuit substrate (18); the digital temperature sensor (17) is partially fitted to the silicon-based heating plate (16); and a second limiting card slot (20) is provided on the heater circuit substrate (18).

6. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The chip clamping module structure (3) comprises a circular spring (22), a pressing device (24), and a fixing member (25); the circular spring (22) is connected to the pressing device (24) via a connecting assembly; the pressing device (24) is arranged on the fixing member (25); and the pressing device (24) is used to push the circular spring (22) to press and clamp the chip (14).

7. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 6, characterized in that: A limit block (23) is provided on the bottom surface of the fixing member (25), and the limit block (23) is used to prevent the pressing device (24) from pushing the circular spring (22) too far, thereby damaging the chip.

8. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The rapid heat dissipation structure (4) is a heat dissipation fan.

9. The handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The power supply module is provided with a plurality of power supply ports which are independent of each other.

10. The use of the handheld variable temperature high-throughput nucleic acid amplification detector according to claim 1, characterized in that: The control module is an application software installed on the mobile phone, which is connected to the detection module via Bluetooth, and can control each detection module individually and add new detection modules as needed.

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